Patent Description:
Companion animals such as dogs and cats frequently require differing diets depending on their life stage (age), size, body composition, and breed. Both dog and cat nutrient requirements can be separated into three different life-stages, based on age: growing dogs (or cats), adult dogs (or cats), and senior dogs (or cats). The latter category, senior dogs (or cats), can be further separated into two stages, which include senior (or mature adult) and super senior (or geriatric). Dogs are further separated into different categories for regular breed dogs versus large-breed dogs.

Essential fatty acids, consisting of omega-<NUM> and omega-<NUM> polyunsaturated fatty acids, are critical nutrients for the health of an animal. These nutrients, however, either cannot be made by animals or cannot be made in sufficient amounts to elicit benefits and therefore must be consumed in an animal's diet. See, e.g., <NPL>). It has previously been postulated that Docosahexaenoic Acid ("DHA"), an omega-<NUM> polyunsaturated fatty acid, is effective in increasing the maze-learning ability and brain functions in aged mice. See, <NPL>).

Rogers discusses the theory of the potential use of antioxidants to slow the deterioration of cognitive function, particularly in the elderly. See <NPL>). In addition, the European Patent Office identified the following prior art documents: <CIT> relates to the use of psyllium seeds or parts of seeds in dry dog food in order to reduce appetite of dogs as well as a process to obtain said dog food. <CIT> discloses a composition and process for controlling postprandial glycemic and/or insulinemic response in companion animals such as geriatric dogs. <CIT> discloses a method and composition to protect an obligate carnivore from a disease of abnormal carbohydrate metabolism by feeding the carnivore a nutritionally balanced diet. <CIT> provides a composition for feline consumption and methods of preparing such compositions, wherein the composition has a moisturized appearance and visually recognizable discrete food particles upon slicing. Laflamme (<NPL>)) discusses nutrition for aging cats and dogs and the importance of body condition by illustrating the effects of aging on nutritional requirements in geriatric animals and the common diet-sensitive conditions in geriatric animals. <CIT> discloses a diet food having effects to reduce body weight and prevent/improve obesity and cardiovascular disorders, diabetes, as well as hyperlipidemia. Backus et al. (<NPL>)) reports on the effect of gonadectomy and high dietary fat in domestic cats. The data provides evidence that a high dietary fat, but not carbohydrate, induces weight gain and congruent increase in insulin, while gonadectomy increases sensitivity to weight gain induced by dietary fat. Rand et al. (<NPL>)) discusses the occurrence of Diabetes Mellitus in Canine and Felines addressing genetic and environmental factors. <CIT> discloses methods for enhancing the quality of life of a senior or super senior animal by feeding the animal a composition comprising at least one omega-<NUM> polyunsaturated fatty acid in combination with other nutrients in amounts effective to enhance altertness, improve vitality, protect cartilage, maintain muscle mass, enhance digestability, improve skin and pelage quality.

Despite the studies and developments relating to improving cognitive abilities, there continues to be a need for methods for enhancing the quality of life of senior animals, as measured by, e.g., enhanced alertness, improved vitality, cartilage protection, maintenance of muscle mass, enhanced digestibility, and improved skin and pelage quality in senior and super senior animals. As previously reported, the super senior pet food composition described herein may be administered to achieve this result. Additionally, we now report herein our surprising discovery that the enhanced quality of life of senior and super senior animals achieved by the administration of the pet food compositions disclosed herein and is reflected at the genomic level. Specifically, as described in detail in the Examples below, gene chip data indicate that the expression of genes that encode proteins associated with several biological pathways such as blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and electron transport are modified, i.e., in general, the majority are beneficially altered through administration to the animal of the super senior pet food compositions described herein.

The invention provides a composition for use in preventing or treating diabetes in a companion animal, wherein the companion animal is a small breed canine that weighs less than <NUM> pounds when an adult and is age greater than <NUM> years, a regular breed canine that weighs less than <NUM> pounds when an adult with age greater than <NUM> years, a large breed canine that weighs more than <NUM> pounds when an adult with age greater than <NUM>, or a feline with age of greater than <NUM> years, wherein the composition comprises at least <NUM>% by weight protein, at least <NUM>% by weight fat, and at least <NUM>% by weight of at least one omega-<NUM> polyunsaturated fatty acid, and wherein the use comprises feeding the composition to the companion animal and wherein the companion animal is a canine or a feline, as defined in the claims.

In an embodiment, the composition comprises at least one omega-<NUM> polyunsaturated fatty acid chosen from the group consisting of docosahexaenoic acid ("DHA") and eicosapentaenoic acid ("EPA"), as defined in the claims.

In yet an additional embodiment, the composition further comprises at least one antioxidant and at least one nutrient selected from the group consisting of choline, manganese, methionine, cysteine, L-carnitine, lysine, and mixtures thereof, as further defined in the claims.

Other and further objects, features, and advantages of the present invention will be readily apparent to those skilled in the art.

It is contemplated that the invention described herein is not limited to the particular methodology, protocols, and reagents described as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention in any way, the scope of the present invention being defined by the claims.

Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods, devices and materials are now described.

In practicing the present disclosure, many conventional techniques in molecular biology may be used. These techniques are well known and are explained in, for example,<NPL>. ); <NPL>); <NPL>. ); <NPL>); <NPL>); <NPL>); and <NPL>).

As used herein and in the appended claims, the singular forms "a", "an", and "the" include plural reference unless the context clearly dictates otherwise.

The terms "senior" or "mature adult" refers to the life-stage of an animal. For small or regular breed canines, the "senior" life stage is from <NUM> to <NUM> years of age. For felines, the "senior" life stage is from <NUM> to <NUM> years of age. For large breed canines, over <NUM> years of age represents "super senior" as described below.

The terms "super senior" or "geriatric" refers to a specific life-stage of an animal. For small or regular breed canines, the super senior stage is any age greater than <NUM> years of age. For large breed canines, the super senior stage is any age greater than <NUM> years of age. For felines, the super senior stage is any age greater than <NUM> years of age.

The term "large breed" canine means a canine that weighs more than <NUM> pounds when an adult.

The term "regular breed" canine means a canine that weighs less than <NUM> pounds when an adult.

The term "small breed" canine means a canine that weighs less than <NUM> pounds when an adult.

The term "super senior pet food composition" refers to any and all of the pet food compositions disclosed herein.

The term "carbohydrate" as used herein includes polysaccharides (e.g., starches and dextrins) and sugars (e.g. sucrose, lactose, maltose, glucose, and fructose) that are metabolized for energy when hydrolyzed. Examples of carbohydrates suitable for inclusion in the compositions disclosed herein include, but are not limited to, com, grain sorghum, wheat, barley, and rice.

The term "antioxidant" means a substance that is capable of reacting with free radicals and neutralizing them. Illustrative examples of such substances include beta-carotene, selenium, coenzyme Q10 (ubiquinone), luetin, tocotri enols, soy isoflavones, <NUM>- adenosylmethionine, glutathione, taurine, N-acetylcysteine, vitamin E, vitamin C, lipoic acid and L-carnitine. Examples of foods containing useful levels of one or more antioxidants include but are not limited to ginkgo biloba, green tea, broccoli, citrus pulp, grape pomace, tomato pomace, carrot spinach, and a wide variety of fruit meals and vegetable meals. It will be understood by one of skill in the art that while units of antioxidants may be provided herein as "ppm", appropriate amounts of antioxidants may also be provided as "IU/kg" where appropriate and customary for a given antioxidant such as, e.g., Vitamin E.

The terms "beneficial change" in gene expression, or gene expression may be "beneficially altered" and like terms refer to a modification in gene expression (e.g., up or down regulation of mRNA levels) such that levels of proteins encoded by the genes may be correspondingly modified such that an associated biological pathway may be more likely to function normally and with less tendency to reflect pathological changes in the pathway that, e.g., may be typical of a super senior animal. Generally, beneficial changes in gene expression relate to improved health and/or reduced propensity for disease in an animal. As used herein, measuring differences in gene expression "levels" and like terms refer to, e.g., characterizing whether expression of a gene is up or down regulated in an animal compared to a control level.

As used herein, "improving" or "enhancing" the quality of life of an animal refers to as an improvement or enhancement in one or more characteristics selected from a group consisting of alertness, vitality, protection of cartilage, maintenance of muscle mass, digestibility, and skin and pelage quality. Additionally, improvement/enhancement in blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and electron transport are also contemplated.

An "improvement" or an "enhancement" in a characteristic or biological pathway refers to a modification in said characteristic or biological pathway such that there is a tendency for the characteristic or pathway to appear and/or function normally and with less tendency to reflect pathological changes in the characteristic or pathway that, e.g., may be typical of a super senior animal.

As used herein, methods to "treat" an animal suffering from a disease or disorder is also meant to encompass methods to prevent and/or to ameliorate the disease or disorder as well.

The present invention provides a composition for use in preventing or treating diabetes in a companion animal, wherein the companion animal is a small breed canine that weighs less than <NUM> pounds when an adult and is age greater than <NUM> years, a regular breen canine that weighs less than <NUM> pounds when an adult whith age greater than <NUM> years, a large breed canine that weighs more than <NUM> pounds when an adult with age greater than <NUM>, or a feline with age of greater than <NUM> years, wherein the composition comprises at least <NUM>% by weight protein, at least <NUM>% by weight fat, and at least <NUM>% by weight omega-<NUM> polyunsaturated fatty acid and wherein the use comprises feeding the composition to the companion animal and wherein the companion animal is a canine or a feline, as defined in the claims. Further disclosed are methods, which are useful for enhancing alertness, improving vitality, protecting cartilage, maintaining muscle mass, enhancing digestibility, and improving skin and pelage quality in a senior or super senior animal. The disclosed methods are also useful for improving in an animal one or more biological pathways selected from the group consisting of blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and the electron transport pathway, such improvements also being reflected in overall beneficial changes at the genomic level. Methods for treating animals suffering from disorders or diseases associated with or related to these biological pathways comprising administering the compositions are also disclosed herein.

Without being bound by theory, the benefits of the invention may be the result of physiological effects from the addition of omega-<NUM> polyunsaturated fatty acids to a senior or super senior animal's diet. Similarly, the antioxidants, choline, and other nutrients may play a role in enhancing a senior or super senior animal's quality of life.

Although the disclosed methods may improve an animal's quality of life by enhancing all of the above described characteristics or improving all of the described biological pathways, it is not necessary to demonstrate substantial improvements in each of the characteristics or pathways to achieve the "enhanced quality of life" as defined herein.

When the compositions are administered to a senior or super senior animal, the animal experiences an enhanced quality of life, e.g., exhibits or experiences one or more of enhanced alertness, improved vitality, protected cartilage, maintained muscle mass, enhanced digestibility, improved skin and pelage quality, as well as improvements in e.g., blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and the electron transport pathway as indicated by overall beneficial changes at the genomic level. Methods for determining these measurements of quality of life are known to skilled artisans. For example, alertness can be measured by various means, including an analysis of metabolism and antioxidant markers, as well as through clinical studies with follow-up questions to participating pet owners. Potential metabolism markers may include ghrelin, GLP-<NUM>, thyroid hormone, and/or growth hormone. Potential markers of antioxidant status may include serum vitamin E, ORAC, glutathione peroxidase, alkanels, and/or cell damage indicators. Further, vitality can be measured by various means, including an analysis of metabolism and antioxidant markers, as well as through clinical studies with follow-up questions to participating pet owners. Similarly, cartilage protection can be measured by various means, including an analysis of arthritis biomarkers. Potential arthritis biomarkers may include type II collagen synthesis, matrix metalloproteinase, osteocalcin, alkaline phosphatase activity, COMP, and fragments of cartilage damage. Muscle mass maintenance can be measured by various means, including an analysis of body composition and digestibility can be measured by various means, including clinical studies with follow-up questions to participating pet owners and animal feeding to determine the percentage of nutrients digested. Skin and pelage quality can be measured by various means, including clinical studies with follow-up questions to participating pet owners. Additionally, as discussed above, improvements in quality of life is also reflected at the genomic level, as evidenced by gene chip data which indicate beneficial changes on the expression of a majority of genes associated with various important biological pathways including blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and protection and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and the electron transport pathway. The identities of these genes are provided in the Examples below.

The methods of the disclosure are useful for enhancing the quality of life of humans and animals, including primates (e.g., monkeys, chimpanzees, etc.), companion animals (e.g., dogs, cats, horses, etc.), farm animals (e.g., goats, sheep, swine, cattle, etc.), laboratory animals (e.g., mice, rats, etc.), birds (e.g., domestic birds such as canaries, parrots, etc. and commercial birds such as chickens, ducks, turkeys, etc.), rodents (e.g., hamsters, guinea pigs, gerbils, rabbits, hedgehogs, ferrets, chinchillas, etc.), and wild, exotic, and zoo animals (e.g., wolves, bears, deer, etc.). In the composition for use of the present invention, the animal is a cat or a dog.

The compositions of the present disclosure are designed to enhance digestibility and improve chewability. Canine and feline foods are typically formulated based on life stage (age), size, body composition, and breed. Thus, some embodiments of the present invention include compositions that are formulated to address specific nutritional differences between regular or small breed dogs, large breed dogs, and cats.

The invention provides a composition containing at least one omega-<NUM> polyunsaturated fatty acid, as defined in the claims. The composition optionally includes foods, supplements, treats, and toys (typically chewable and consumable toys). The composition is fed to the designated animals over a period of time that is long enough to effectuate the improved quality of life. In one embodiment, the composition is provided to the animal for at least thirty days.

The compositions for use of the present invention generally have an omega-<NUM> polyunsaturated fatty acid content of at least <NUM>% (or from <NUM> % to <NUM>%, or from <NUM>% to <NUM>%) by weight on a dry matter basis, as defined in the claims. In some embodiments, the omega-<NUM> polyunsaturated fatty acid is DHA. In other embodiments, the omega-<NUM> polyunsaturated fatty acid is EPA. In still other embodiments, the omega-<NUM> polyunsaturated fatty acid comprises a mixture of DHA and EPA.

In some embodiments, the composition containing omega-<NUM> polyunsaturated fatty acid is a food. Although both liquid and solid foods are provided, solid foods are typically preferred. Foods include both dry foods and wet foods. Some of the non-polyunsaturated fatty acid components of the food, and their preferred proportions, include those listed in Table <NUM>.

In another embodiment, this invention provides a composition for use in preventing or treating diabetes as further defined in the claims and specifically for a senior or super senior small or regular breed canine. The use comprises feeding the canine a composition comprising:.

In another embodiment, this invention provides a composition for use in preventing or treating diabetes as further defined in the claims and specifically for a senior or super senior large breed canine. The use comprises feeding the canine a composition comprising:.

In another embodiment, this invention provides a composition for use in preventing or treating diabetes as further defined in the claims and specifically for a senior or super senior feline. The use comprises feeding the feline a composition comprising:.

In a further embodiment, this invention provides a composition for use in preventing or treating diabetes as defined in the claims in a senior or super senior animal comprising feeding the animal (e.g., small, regular or large breed canine or feline, as the case may be), wherein the composition comprises the components as indicated in Table 1A below:.

The compositions for use disclosed herein optionally further comprise at least one nutrient selected from the group consisting of manganese, methionine, cysteine, mixtures of methionine and cysteine, L-carnitine, lysine, and arginine. Specific preferred amounts for each component in a composition will depend on a variety of factors including, for example, the species of animal consuming the composition; the particular components included in the composition; the age, weight, general health, sex, and diet of the animal; the animal's consumption rate, and the like. Thus, the component amounts may vary widely, and may even deviate from the proportions given herein.

The omega-<NUM> fatty acids may be obtained from a variety of sources. One convenient source is fish oils from, for example, menhaden, mackerel, herring, anchovy, and salmon. DHA and EPA are typical fatty acids present in such fish oils, and, together often make up a significant portion of the oil, such as from <NUM>% to <NUM>% of the oil.

When the composition is an animal food, vitamins and minerals preferably are included in amounts required to avoid deficiency and maintain health. These amounts are readily available in the art. The National Research Council (NRC), for example, provides recommended amounts of such ingredients for farm animals. See, e.g., <NPL>), <NPL>), <NPL>), <NPL>). The American Feed Control Officials (AAFCO), for example, provides recommended amounts of such ingredients for dogs and cats. See <NPL>). Examples of vitamins useful as food additives include vitamin A, BI, B2, B6, B12, C, D, E, K, H (biotin), K, folic acid, inositol, niacin, and pantothenic acid. Examples of minerals and trace elements useful as food additives include calcium, phosphorus, sodium, potassium, magnesium, copper, zinc, chloride, and iron salts.

The compositions for use of the present invention may further contain other additives known in the art. Preferably, such additives are present in amounts that do not impair the purpose and effect provided by the invention. Examples of additives include, for example, substances with a stabilizing effect, processing aids, substances that enhance palatability, coloring substances, and substances that provide nutritional benefits.

Stabilizing substances include, for example, substances that tend to increase the shelf life of the composition. Potentially suitable examples of such substances include, for example, preservatives, antioxidants, synergists and sequestrants, packaging gases, stabilizers, emulsifiers, thickeners, gelling agents, and humectants. Examples of emulsifiers and/or thickening agents include, for example, gelatin, cellulose ethers, starch, starch esters, starch ethers, and modified starches.

Additives for coloring, palatability ("pal enhancers"), and nutritional purposes include, for example, colorants (e.g., iron oxide, such as the red, yellow, or brown forms); sodium chloride, potassium citrate, potassium chloride, and other edible salts; vitamins; minerals; and flavoring. Such additives are known in the art. See, e.g., <CIT>. See also, <CIT>. Flavorants include, for example, dairy product flavorants (e.g., milk or cheese), meat flavorants (e.g., bacon, liver, beef, poultry, or fish), oleoresin, pinacol, and the various flavorants identified in the trade by a FEMA (Flavor Extract Manufacturers Association) number. Flavorants help provide additional palatability, and are known in the art. See, e.g., <CIT>. See also, <CIT>. See also, <CIT>. See also, <CIT>. See also, <CIT>. The concentration of such additives in the composition typically may be up to <NUM>% by weight. In some embodiments, the concentration of such additives (particularly where such additives are primarily nutritional balancing agents, such as vitamins and minerals) is from <NUM>% to <NUM>% by weight. In some embodiments, the concentration of such additives (again, particularly where such additives are primarily nutritional balancing agents) is from <NUM>% to <NUM>% by weight.

Supplements include, for example, a feed used with another feed to improve the nutritive balance or performance of the total. Supplements include compositions that are fed undiluted as a supplement to other feeds, offered free choice with other parts of an animal's ration that are separately available, or diluted and mixed with an animal's regular feed to produce a complete feed. The AAFCO, for example, provides a discussion relating to supplements in the American Feed Control Officials, Inc. Official Publication, p. <NUM> (<NUM>). Supplements may be in various forms including, for example, powders, liquids, syrups, pills, encapsulated compositions, and the like.

Treats include, for example, compositions that are given to an animal to entice the animal to eat during a non-meal time. Treats for canines include, for example, dog bones. Treats may be nutritional, wherein the composition comprises one or more nutrients, and may, for example, have a composition as described above for food. Non-nutritional treats encompass any other treats that are non-toxic.

Toys include, for example, chewable toys. Toys for dogs include, for example, artificial bones. There is a wide range of suitable toys currently marketed. See, e.g., <CIT> (and references disclosed in <CIT>). See also, e.g., <CIT> (and references disclosed in <CIT>). The invention optionally provides both partially consumable toys (e.g., toys comprising plastic components) and fully consumable toys (e.g., rawhides and various artificial bones). It should be further recognized that this invention optionally provides toys for for dog or cat use.

A "food" is a nutritionally complete diet for the intended recipient animal (e.g., domestic cat or domestic dog). A "nutritionally complete diet" is a diet that includes sufficient nutrients for maintenance of normal health of a healthy animal on the diet. The compositions of this invention are not intended to be restricted by any specific listing of proteinaceous or fat ingredients or product form. The compositions can be prepared in, for example, a dry, canned, wet, or intermediate moisture form using conventional pet food processes. In some embodiments, the moisture content is from <NUM>% to <NUM>% of the total weight of the composition. In other embodiments, the moisture content is from <NUM>% to <NUM>% of the total weight of the composition.

In preparing a composition for use according to the present invention, any ingredient (e.g., fish oil) generally may, for example, be incorporated into the composition during the processing of the formulation, such as during and/or after mixing of other components of the composition. Distribution of these components into the composition can be accomplished by conventional means. In one embodiment, ground animal and poultry proteinaceous tissues are mixed with the other ingredients, including fish oils, cereal grains, other nutritionally balancing ingredients, special-purpose additives (e.g., vitamin and mineral mixtures, inorganic salts, cellulose and beet pulp, bulking agents, and the like); and water that is sufficient for processing is also added. These ingredients preferably are mixed in a vessel suitable for heating while blending the components. Heating of the mixture may be effected using any suitable manner, such as, for example, by direct steam injection or by using a vessel fitted with a heat exchanger. Following the addition of the last ingredient, the mixture is heated to a temperature range of from <NUM> (<NUM>°F) to <NUM> (<NUM>°F). In some embodiments, the mixture is heated to a temperature range of from <NUM> (<NUM>°F) to <NUM> (<NUM>°F). Temperatures outside these ranges are generally acceptable, but may be commercially impractical without use of other processing aids. When heated to the appropriate temperature, the material will typically be in the form of a thick liquid. The thick liquid is filled into cans. A lid is applied, and the container is hermetically sealed. The sealed can is then placed into conventional equipment designed to sterilize the contents. This is usually accomplished by heating to temperatures of greater than <NUM> (<NUM>°F) for an appropriate time, which is dependent on, for example, the temperature used and the composition.

Compositions for use of the present invention can be prepared in a dry form using conventional processes. In one embodiment, dry ingredients, including, for example, animal protein sources, plant protein sources, grains, etc., are ground and mixed together. Moist or liquid ingredients, including fats, oils, animal protein sources, water, etc., are then added to and mixed with the dry mix. The mixture is then processed into kibbles or similar dry pieces. Kibble is often formed using an extrusion process in which the mixture of dry and wet ingredients is subjected to mechanical work at a high pressure and temperature, and forced through small openings and cut off into kibble by a rotating knife. The wet kibble is then dried and optionally coated with one or more topical coatings which may include, for example, flavors, fats, oils, powders, and the like. Kibble also can be made from the dough using a baking process, rather than extrusion, wherein the dough is placed into a mold before dry-heat processing.

The compositions are also designed to be easier to chew. Canine and feline foods are typically formulated based on life stage (age), size, body composition, and breed. In the compositions for use of this invention, some embodiments of the compositions address specific nutritional differences between super senior regular or small breed dogs, large breed dogs, and cats.

All percentages expressed herein are on a weight by dry matter basis unless specifically stated otherwise.

As noted previously, this disclosure provides, in part, a method for enhancing the quality of life of an animal. The method comprises feeding a senior or super senior animal a composition in an amount effective to enhance alertness, improve vitality, protect cartilage, maintain muscle mass, enhance digestibility, and improve skin and pelage quality. Additionally, we now report herein our surprising discovery that the enhanced quality of life of an animal achieved by administration of the compositions of the present disclosure is reflected at the genomic level. While it may be that a change in expression of any one gene disclosed in the tables presented below may result in beneficial or deleterious biological effects, the data presented herein indicate that, overall, the observed expression profiles are consistent with the beneficial biological effects seen in vivo after administration of the diets disclosed herein. Specifically, gene chip data indicate that the expression of genes that encode proteins associated with or related to several biological pathways such as blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and electron transport are, for the most part, beneficially altered through administration to the animal of compositions described herein. Thus, the disclosure also relates to methods of measuring or characterizing the enhancement in the quality of life of an animal, particularly a senior or super senior animal, fed a composition described herein by quantitating the gene expression levels of one or more genes selected from a group consisting of those disclosed in Tables <NUM>-<NUM> in said animal and comparing said levels in the animal to levels in the animal prior to administration of the feed composition. Quantitation of gene expression may be carried out in numerous ways familiar to one of skill in the art and include such techniques as RT PCR as well as gene chip assays and Northern blotting. Thus, it is contemplated herein that the expression levels detected may be used, for example, in methods to measure enhancement in the quality of life of an animal as disclosed herein.

In another aspect, the present disclosure relates to kits which comprise:.

It will be appreciated that in any such kit, (a), (b), (c) or (d) may comprise a substantial component. The manufacture of kits as described herein and components thereof (e.g., antibody production) may be achieved according to conventional methods.

It is contemplated herein that modulating the expression levels of the genes disclosed herein may have therapeutic value with regard to the treatment of diseases or disorders associated with the various biological pathways. Such determination may be made on a gene by gene basis without undue experimentation, for example, by assessing expression levels in tissues as well as in blood samples, or by assaying expression levels in vitro in cells or cell lines relevant to particular disease states and suitable for such experimentation. In vivo models of disease might also be utilized in such experimentation. The nature of these and other suitable additional assays would be familiar to one of skill in the art. Thus, based on the genomic data disclosed herein, the disclosure also relates to methods to enhance the quality of life of an animal by modulating the expression level of one or more genes listed on Tables <NUM>-<NUM> (i.e. up or down regulation as indicated therein) in an animal in order to mimic the pattern of expression seen in vivo after administration of the pet food compositions.

Modulation of gene expression levels may be achieved through the use of known modulators of gene expression suitable for administration in vivo, including, but not limited to, ribozymes, antisense oligonucleotides, triple helix DNA, RNA aptamers and/or double stranded RNA directed to an appropriate nucleotide sequence of a gene of interest. These inhibitory molecules may be created using conventional techniques by one of skill in the art without undue burden or experimentation. For example, modification (e.g. inhibition) of gene expression may be obtained by designing antisense molecules, DNA or RNA, to the control regions of the genes discussed herein, i.e. to promoters, enhancers, and introns. For example, oligonucleotides derived from the transcription initiation site, e.g., between positions -<NUM> and +<NUM> from the start site may be used. Notwithstanding, all regions of the gene may be used to design an antisense molecule in order to create those which gives strongest hybridization to the mRNA and such suitable antisense oligonucleotides may be produced and identified by standard assay procedures familiar to one of skill in the art.

Similarly, inhibition of gene expression may be achieved using "triple helix" base-pairing methodology. Triple helix pairing is useful because it causes inhibition of the ability of the double helix to open sufficiently for the binding of polymerases, transcription factors, or regulatory molecules. Recent therapeutic advances using triplex DNA have been described in the literature (<NPL>. These molecules may also be designed to block translation of mRNA by preventing the transcript from binding to ribosomes.

Ribozymes, enzymatic RNA molecules, may also be used to modulate gene expression by catalyzing the specific cleavage of RNA. The mechanism of ribozyme action involves sequence-specific hybridization of the ribozyme molecule to complementary target RNA, followed by endonucleolytic cleavage. Examples which may be used include engineered "hammerhead" or "hairpin" motif ribozyme molecules that can be designed to specifically and efficiently catalyze endonucleolytic cleavage of gene sequences.

Specific ribozyme cleavage sites within any potential RNA target are initially identified by scanning the target molecule for ribozyme cleavage sites which include the following sequences: GUA, GUU and GUC. Once identified, short RNA sequences of between <NUM> and <NUM> ribonucleotides corresponding to the region of the target gene containing the cleavage site may be evaluated for secondary structural features which may render the oligonucleotide inoperable. The suitability of candidate targets may also be evaluated by testing accessibility to hybridization with complementary oligonucleotides using ribonuclease protection assays.

Ribozyme methods include exposing a cell to ribozymes or inducing expression in a cell of such small RNA ribozyme molecules (<NPL>; <NPL>). Intracellular expression of hammerhead and hairpin ribozymes targeted to mRNA corresponding to at least one of the genes discussed herein can be utilized to inhibit protein encoded by the gene.

Ribozymes can either be delivered directly to cells, in the form of RNA oligonucleotides incorporating ribozyme sequences, or introduced into the cell as an expression vector encoding the desired ribozymal RNA. Ribozymes can be routinely expressed in vivo in sufficient number to be catalytically effective in cleaving mRNA, and thereby modifying mRNA abundance in a cell (<NPL>). In particular, a ribozyme coding DNA sequence, designed according to conventional, well known rules and synthesized, for example, by standard phosphoramidite chemistry, can be ligated into a restriction enzyme site in the anticodon stem and loop of a gene encoding a tRNA, which can then be transformed into and expressed in a cell of interest by methods routine in the art. Preferably, an inducible promoter (e.g., a glucocorticoid or a tetracycline response element) is also introduced into this construct so that ribozyme expression can be selectively controlled. For saturating use, a highly and constituently active promoter can be used. tDNA genes (i.e., genes encoding tRNAs) are useful in this application because of their small size, high rate of transcription, and ubiquitous expression in different kinds of tissues. Therefore, ribozymes can be routinely designed to cleave virtually any mRNA sequence, and a cell can be routinely transformed with DNA coding for such ribozyme sequences such that a controllable and catalytically effective amount of the ribozyme is expressed. Accordingly the abundance of virtually any RNA species in a cell can be modified or perturbed.

Ribozyme sequences can be modified in essentially the same manner as described for antisense nucleotides, e.g., the ribozyme sequence can comprise a modified base moiety.

RNA aptamers can also be introduced into or expressed in a cell to modify RNA abundance or activity. RNA aptamers are specific RNA ligands for proteins, such as for Tat and Rev RNA (<NPL>) that can specifically inhibit their translation.

Gene specific inhibition of gene expression may also be achieved using conventional RNAi technologies. Numerous references describing such technologies exist and include, for example, <CIT>;<NPL>); <NPL>).

Antisense molecules, triple helix DNA, RNA aptamers and ribozymes may be prepared by any method known in the art for the synthesis of nucleic acid molecules. These include techniques for chemically synthesizing oligonucleotides such as solid phase phosphoramidite chemical synthesis. Alternatively, RNA molecules may be generated by in vitro and in vivo transcription of DNA sequences encoding the genes discussed herein. Such DNA sequences may be incorporated into a wide variety of vectors with suitable RNA polymerase promoters such as T7 or SP6 according to conventional methods. Alternatively, cDNA constructs that synthesize antisense RNA constitutively or inducibly can be introduced into cell lines, cells, or tissues using methods familiar to one of skill in the art. Vectors may be introduced into cells or tissues by many available means, and may be used in vivo, in vitro or ex vivo. For ex vivo therapy, vectors may be introduced into stem cells taken from an animal and clonally propagated for autologous transplant back into that same animal. Delivery by transfection and by liposome injections may be achieved using methods that are well known in the art.

The disclosure also includes a method to identify an animal that might benefit from feeding a composition as disclosed herein comprising measuring the gene expression levels of any one or more genes listed in Tables <NUM>-<NUM> in said animal and comparing said levels to the gene expression levels seen in Tables <NUM>-<NUM> wherein an animal with levels different than those seen in Tables <NUM>-<NUM> (e.g., up regulated versus down regulated) would be identified as potentially benefiting from feeding a composition of the present disclosure.

It is also contemplated herein that the disclosurerelates to methods for treating an animal suffering from disorders or disease associated with or relating to any one of more of the following biological pathways: blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and electron transport comprising administering to the animal a composition of the present disclosure.

The terms "comprise", "comprises", and "comprising" are to be interpreted inclusively rather than exclusively.

Unless defined otherwise, all technical and scientific terms and any acronyms used herein have the same meanings as commonly understood by one of ordinary skill in the art in the field of the invention.

In the specification there have been disclosed typical preferred embodiments of the invention and, although specific terms are employed, they are used in a generic and descriptive sense only and not for purposes of limitation, the scope of the invention being set forth in the following claims. Many modifications and variations are possible in light of the above teachings.

This invention can be further illustrated by the following examples of preferred embodiments thereof, although it will be understood that these examples are included merely for purposes of illustration and are not intended to limit the scope of the invention unless otherwise specifically indicated.

A composition formulated for senior or super senior regular or small breed canines is described in Table <NUM>.

A composition formulated for senior or super senior large breed canines is described in Table <NUM>.

A composition formulated for senior or super senior felines is described in Table <NUM>.

To further characterize the nutritional benefits of the super senior pet food compositions, gene expression profiles from animals fed the compositions compared to control animals are assayed and the results are described in detail below.

Blood samples are drawn from <NUM> Beagles according to conventional methods before and after feeding for <NUM> days on Super Senior K9 diet (a total of <NUM> samples). Each sample taken after the <NUM> day trial is compared to its own control.

<NUM> canine blood, heparin or EDTA tubes, Hank's Balanced Salt Solution (Gibco <NUM>-<NUM>),HEPES buffer (Gibco <NUM>-<NUM>), Accu-Paque (Accurate Chemical & Scientific Corp AN3100).

Transfer pipettes (VWR <NUM>-<NUM>), <NUM> centrifuge tubes w/ caps,<NUM>" Pasteur pipettes,<NUM> microcentrifuge tubes (VWR <NUM>-<NUM>),centrifuge tube racks, microcentrifuge tube rack, waste container, Beckman Coulter Allegra 25R Centrifuge, SN AJC01J015Eppendorf Centrifuge, 5417C.

Hank's Balanced Salt Solution (HBSS) w/ <NUM> HEPES buffer solution is made by adding <NUM> of HEPES buffer solution to a <NUM> bottle of HBSS. Hank's Balanced Salt Solution and Accu-Paque need to be removed from the refrigerator and placed at room temperature at least <NUM> minutes before beginning the lymphocyte isolation. Both solutions should be place back in the refrigerator (<NUM>) immediately following their use.

Deionized H<NUM>O, Absolute ethanol (Sigma E7023), RNA Storage Solution (Ambion <NUM>),RNase Zap® (Ambion <NUM>),Buffer RLT, Buffer RW1 and Buffer RPE (provided in the RNeasy Mini Kit).

RNeasy Mini Kit (Qiagen <NUM>), QIAshredder spin columns (Qiagen <NUM>),P1000 Pipetman pipette (Rainin), P200 Pipetman pipette (Rainin), <NUM>-<NUM>µl filtered pipette tips (USA Scientific <NUM>-<NUM>),<NUM>-<NUM>µl filtered pipette tips (USA Scientific <NUM>-<NUM>), sterile transfer pipettes (VWR <NUM>-<NUM>),<NUM> sterile solution basin (VWR <NUM>-<NUM>),<NUM> waste containers (one for liquid, one for tips/pipettes),<NUM> sterile microcentrifuge tubes (VWR <NUM>-<NUM>), Microcentrifuge tube rack, permanent marker, Eppendorf Microcentrifuge, model #5417C.

Ovation TM Biotin System v1. <NUM> for probe preps.

User Guide (Cat#D01002, version <NUM>/<NUM>/<NUM>, NuGEN Technologies, Inc). The experimental procedure is followed as described in the user guide. All probe preparation starts with <NUM> ng of total RNA.

The Genechips used for the test is the Canine Genome <NUM> Array (Affymetrix). This Genechip contains <NUM>,<NUM> probe sets. Detailed sequence information for each unique probe identification number is available from the manufacturer.

Normalization is performed using MAS <NUM> provided in GCOS Affymetrix software (version <NUM>). Expression levels for the genes analyzed are indicated on the tables included in the examples below, where an upward facing arrow refers to "up regulation" or increase and a downward facing arrow indicates "down regulation" in gene expression. Similarly, in some tables, upward or downward facing arrows also indicate increases or decreases in activity of certain proteins involved in a particular pathway, and are otherwise self explanatory.

<NUM>,<NUM> genes are selected for further analysis based on their "present" calls in at least <NUM> out of <NUM> samples.

Results of the gene chip analysis indicate that <NUM> genes are differentially expressed between the control and Super Senior diet treated groups. The expression levels of these <NUM> genes are statistically significant when grouped by 'diet'; using a parametric test where the variances is not assumed to be equal (Welch t-test). The p-value cutoff is <NUM> with no multiple testing correction. Under those selection criteria only about <NUM> genes would be expected to pass the restriction by chance. The genomic data is discussed in detail below.

Based on an analysis of the gene chip data, at the P <<NUM> level, <NUM>,<NUM> genes changed compared to control expression levels (<NUM> were up regulated and the rest down regulated). At the P < <NUM> level, data indicate that <NUM> genes are down regulated in beagles fed the super senior food. Nine of these down regulated genes are identified as related to the inflammatory and pain response. Down regulation of these genes may be predicted to result in pain relief, cartilage protection (less damage) and reduction in inflammatory responses. The compositions disclosed herein may be part of a therapeutic regimen to treat animals suffering from pain and/or inflammatory diseases. These genes and their putative role in inflammation and pain response are provided below in Tables <NUM>-<NUM>.

At the P < <NUM> and P < <NUM> level, <NUM> genes are identified to be related to heart health through regulation of the eicosanoid pathway and blood coagulation pathway. The genes are responsible for blood coagulation through platelet activation and aggregation. The down regulation of these genes through nutrition can prevent inappropriate blood clotting which may result in heart or brain related disorders. The compositions of the present disclosure may be part of a therapeutic regimen to treat animals suffering from disorders or diseases of the blood, heart or brain. These genes and their putative role in vivo are described in Tables <NUM> and <NUM> below.

Ten down regulated genes are identified as related to body composition through regulation of bone and muscle. The genes spare muscle and bone deterioration by reducing nitric oxide production and glucocorticoid degradation of muscle. Down regulation of these genes results in a decrease in nitric oxide production and glucocorticoid response. The compositions disclosed herein may be part of a therapeutic regimen to treat animals suffering from diseases or disorders associated with or relating to muscle or bone. These genes and their putative role in muscle and bone regulation are detailed in Tables <NUM> and <NUM> below.

Eleven genes are identified that are related to DNA damage/protection and neural function. With regard to the latter, the genes identified are important for rebound potentiation; they are believed to have a potential role in motor learning. Interestingly, of these genes, all were down regulated except for of gamma-aminobutyric acid (GABA) A receptor, gamma <NUM> which was up regulated. The compositions disclosed herein may be part of a therapeutic regimen to treat animals suffering from diseases or disorders associated with or relating to DNA damage/protection and neural function. The identity of these genes and their putative role in DNA damage/protection and neural function are described in Tables <NUM> and <NUM> below.

Twenty four genes associated with glucose metabolism are down regulated in animals fed the super senior diet which would suggest that these animals are utilizing fat (fat oxidation) instead of glucose as a fuel source. The compositions disclosed herein may be part of a therapeutic regime in diabetic animals and/or for obesity prevention or treatment in an animal. These down regulated genes are identified and their putative role in glucose metabolism described in detail below in Tables <NUM> and <NUM>.

Claim 1:
A composition for use in preventing or treating diabetes in a companion animal, wherein the companion animal is a small breed canine that weighs less than <NUM> pounds when an adult and is age greater than <NUM> years, a regular breed canine that weighs less than <NUM> pounds when an adult with age greater than <NUM> years, a large breed canine that weighs more than <NUM> pounds when an adult with age greater than <NUM>, or a feline with age of greater than <NUM> years, wherein the composition comprises:
at least <NUM> % by weight protein;
at least <NUM> % by weight fat; and
at least <NUM> % by weight of at least one omega-<NUM> polyunsaturated fatty acid,
wherein the use comprises feeding the composition to the companion animal, and wherein the companion animal is a canine or a feline.