# Patent

## Title
Methods for improving health in canines

## Abstract

A method of improving health in a canine includes administering to the canine a nutritional supplement comprising an amino acid secretagogue composition, which stimulates the pituitary gland of the canines to produce growth hormone. The nutritional supplement may be administered orally. The nutritional supplement may comprise L-arginine hydrochloride, Oxo-proline, L-lysine hydrochloride, and cysteine. When desired, the nutritional supplement may consist essentially of L-arginine hydrochloride, Oxo-proline, L-lysine hydrochloride, N-acetyl-L-cysteine, L-glutamine, and schizonepeta powder.

## Background
TECHNICAL FIELD
Embodiments of the present disclosure generally relate to methods and nutritional supplements for improving health in canines.
BACKGROUND
Growth hormone is an anabolic hormone capable of promoting linear growth, weight gain and whole body nitrogen retention in animals. Growth hormone is produced in somatotroph cells of the anterior pituitary gland of animals and secreted throughout life. It is mainly controlled in the brain by two hypothalamic peptides: growth hormone releasing hormone (GHRH), which stimulates its secretion and synthesis; and somatostatin, which inhibits them. In human beings and other mammals, regulated expression of the growth hormone pathway is considered essential for optimal linear growth, as well as homeostasis of carbohydrate, protein and fat metabolism.
As canines age, their growth hormone levels decrease considerably resulting in a loss of skeletal muscle mass, osteoporosis, increased fat deposition, decreased lean body mass, and other disorders. Studies in human beings and other mammals have demonstrated that the recombinant growth hormone therapy may address the reduction of growth hormone. However, the recombinant growth hormone therapy for canines is limited and has proved to be inefficient.
U.S. Pat. No. 7,361,642, issued on Apr. 22, 2008 to Draghia-Akli, discloses a method for increasing growth hormone in canines. A canine-specific growth hormone releasing hormone (dGHRH) or a nucleic acid molecule that encodes the dGHRH is administered to canines to increase the level of growth hormone secretion. The canines treated with specific growth hormone releasing hormone show increased insulin-like growth factor I, enhanced red blood cells production and hemoglobin concentration, and improved protein metabolism.
Journal of Orthopedic Research 15:519-527 (1997) discloses that a growth hormone secretagogue, MK-0677, elevates levels of serum insulin-like growth factor-I in canines, which in turn increases the size and strength of the quadriceps muscle in canines during remobilization.
U.S. Publication No. 2008/0119819, published on May 22, 2008 to Hojby, discloses that an administration of growth hormone enhances bone growth factors and fracture healing in canines.
It has also been reported that for adult canines of both sexes, exogenous growth hormone induces a marked increase in new bone formation. (Calcified Tissue Research 10(1):1-13 (1972)).
Other studies have shown that in the canines that were given growth hormone, the quantity of protein in the blood plasma is increased, and the plasma protein pattern is altered. (Endocrinology 53: 134-162 (1953)). Additionally, it is known that growth hormone treatment improves protein metabolism in canines.
SUMMARY OF THE DISCLOSURE
The present disclosure includes a nutritional supplement and a method of using the same for improving health in canines. The nutritional supplement is an amino acid secretagogue composition, which stimulates the pituitary gland of canines to release growth hormone.
A particular embodiment of the present disclosure relates to a nutritional supplement for improving health in canines, the nutritional supplement comprising L-arginine, Oxo-proline, and L-lysine.
Another particular embodiment relates to a nutritional supplement for improving health in canines, the nutritional supplement includes L-arginine hydrochloride, Oxo-proline, L-lysine hydrochloride, N-acetyl-L-cysteine, L-glutamine, and schizonepeta powder.
Other embodiments are drawn to methods of increasing growth hormone in canines, the methods including administering the disclosed nutritional supplement to a canine. In particular embodiments, the method includes orally administering the disclosed nutritional supplement to a canine.


DETAILED DESCRIPTION OF THE DISCLOSURE
The present disclosure relates to methods and nutritional supplements for improving health in canines. The nutritional supplement is an amino acid secretagogue composition, which stimulates the pituitary gland in a canine to produce growth hormone. Increased production of growth hormone in canines may prevent the loss of skeletal muscle mass, osteoporosis, increased fat deposition, decreased lean body mass, and other disorders. The supplement of the present disclosure may work as a dietary supplement by assisting the canine's ability to secrete growth hormone naturally in a manner which is safe and effective.
A particular embodiment of the present disclosure relates to a nutritional supplement for improving health of canines, the nutritional supplement including L-lysine, L-arginine, oxo-proline, and at least one of cysteine and glutamine. The supplement may additionally include both cysteine and glutamine and/or schizonepeta powder. In particular embodiments, a functional dosage may include L-arginine at a level between 0.1-6 mmol, oxo-proline at a level between 0.1-8 mmol, and/or the L-lysine in an amount between 0.1-12 mmol. The cysteine and/or glutamine may be contained at a level between 0.001-6 mmol. The cysteine may be n-acetyl L-cysteine, and the glutamine may be L-glutamine. The amino acids may be delivered as non-toxic salts thereof, effective complexes thereof, stable chelates thereof, active esters thereof, functional derivatives thereof, or mixtures thereof, which are effective to increase growth hormone levels in canines. The nutritional supplement may be present in an amount of 2.9 grams. Dosages are generally calculated in the range of about 20-50 gms per kg of body weight of the canine. The nutritional supplement may be administered to canines orally. The supplement may be in any acceptable and known oral formulation including, but are not limited to, powder, tablet, capsule, or liquid form.
Another particular embodiment relates to a nutritional supplement for improving health of canines, the nutritional supplement includes L-lysine HCl, L-arginine HCl, oxo-proline, N-acetyl L-cysteine, L-glutamine, and schizonepeta (aerial parts) powder. In particular embodiments, a functional dosage may include L-arginine HCl at a level between 0.1-6 mmol, and oxo-proline at a level between 0.1-8 mmol, and/or the L-lysine HCl in an amount between 0.1-12 mmol. The n-acetyl L-cysteine and/or L-glutamine may be contained at a level between 0.001-6 mmol. In another particular embodiment, a functional dosage may include L-arginine HCl at a level between 2.5-4.5 mmol, oxo-proline between 4-6 mmol, and/or the L-lysine HCl in an amount between 7-9 mmol. The n-acetyl L-cysteine and/or L-glutamine may be contained at a level between 0.001-0.5 mmol. The nutritional supplement may be administered to canines orally. The supplement may be in any acceptable and known oral formulation including, but are not limited to, powder, tablet, capsule, or liquid form.
Other embodiments are drawn to methods of increasing growth hormone in canines, the methods including administering the disclosed nutritional supplement to a canine. In particular embodiments, the method includes orally administering the disclosed nutritional supplement to a canine. The amount of dosage and frequency of administration may be varied according to many factors including, but are not limited to, canine breeds, ages (e.g., puppy, adult, elderly) and health of the canines.
The disclosed nutritional supplements may be administered to a canine to improve health, including by: increasing insulin-like growth factor I, enhancing red blood cells production and hemoglobin concentration, boosting protein metabolism, improving bone growth factors and fracture healing, increasing new bone formation, enhancing protein metabolism, or combinations thereof.
In accordance with the “consist essentially of” and “consisting essentially of” language, the nutritional supplement is essentially limited to the aforementioned ingredients and does not include any additional active ingredients intended to add nutritional content (e.g., vitamins, minerals, etc.), but may include additional ingredients not intended to add nutritional content such as ingredients intended to fulfill a non-nutritional purpose (e.g., coloring, fillers, flavoring, an ingredient for maintaining the structural form, etc.).
Each ingredient of the nutritional supplement may be prepared in accordance with any method known to one of ordinary skill in the art. Alternatively, each ingredient may be obtained in a fully prepared from a commercially available source.
The nutritional supplement may be in any suitable oral administration form, including but not limited to: a chewable form, a liquid form, a spray form, a capsule form, a suppository form, dissolvable wafer, and a powder form.
Irrespective of the structural form of the nutritional supplement, the ingredients of the nutritional supplement may be distributed homogeneously or non-homogeneously within the nutritional supplement.
The nutritional supplement may be administered to a canine on a regular basis, such as a weekly or monthly intake at a dosage tailored to the canine's needs; i.e., the nutritional supplement may be taken regularly as multiples (1×, 2×, etc.) of the structural units (pills, tablets, capsules, etc.) in accordance with the needs of the canine. For example, an elderly canine is likely to need higher daily doses than does a young canine. Alternatively, the nutritional supplement may be administered on an as-needed basis at a dosage tailored to the canine's needs, such as administering the nutritional supplement to the canine that is recovering from bone fracture. Veterinarian counseling may be beneficial for arriving at a desirable or optimal dosage tailored to the canine's needs.
The combination of types of amino acids, mass ranges, and specific formulations may be selected to be synergistically balanced and of adequate quantity to achieve the desired physiological effect, namely, growth hormone release. Improper combinations of the amino acids may be ineffective. The component amino acids are synergistic in the sense that several of them when combined together, synergistically stimulate the release of human growth hormone. The combination may also be chosen to reduce or inhibit chemical combination or reaction between the amino acids.
EXAMPLES
Example 1
The short-term effects of the disclosed supplement on the growth hormone level in canine are tested two hours after administration. The canine subjects are administered the capsules of supplement (SeroVitalTM) or an identical looking placebo. SeroVitalTM is a blend of l-lysine HCl, l-arginine HCL, oxo-proline, N-acetyl-l-cysteine, l-glutamine, and schizonepeta (aerial parts) powder. Canine subjects are administered approximately 35 gms per kg of body weight. Blood is drawn at 15, 30, 60 and 90 and 120 minutes for assay. The growth hormone level is measured at each time point using the Siemens Immulite 2000 (intra-assay CV was 3.72%, inter-assay CV was 5.70%, and the detection limit for GH was 0.05 ng/ml). The 15 and 120 minute time points are additionally assayed for triiodothyronine (T3) as informative for mechanistic investigations.
The mean growth hormone in the canine subjects are increased more than 600% after the administration of supplement from 0.17 at baseline to 1.33 ng/ml at 120 minutes, while there is are no increases observed in the mean growth hormone of the canine subjects after the administration of placebo.
The growth hormone levels from baseline to 120 minutes (the growth hormone level at 120 minutes minus the growth hormone level at 0 minute), are increased about two fold when compared to the administration of a placebo. Overall, 120 minutes after the administration of the supplement, the growth hormone levels are significantly higher in absolute levels or by AUC.
Furthermore, the canines that were administered SeroVitalTM will exhibit a deceased reduction in T3 by nearly one-half over the same time period, compared to the canines administered with the placebo. These results affirm that somatostatin inhibition plays a mechanistic role in the ability of SeroVitalTM to induce significant increases in serum growth hormone levels in canine subjects.
These findings demonstrate that a specialized low-dose amino acid supplement can significantly increase short-term growth hormone levels in canines.
Example 2
The effect of the nutritional supplement on endurance and fat metabolism in canines are investigated. Each canine subject is measured for baseline weight, height, body fat percent and resting metabolic rate. Then, each canine subject is administered with the supplement SeroVitalTM, which is a blend of l-lysine HCl, l-arginine HCl, oxo-proline, N-acetyl-l-cysteine, l-glutamine, and schizonepeta (aerial parts) powder for 2 weeks before being measured again for weight, height, body fat percent and resting metabolic rate. The measurements before and after the administration of the supplement are analyzed.
Mean VO2max of the canine subjects increases compared to baseline. After the period of supplementation with SeroVitalTM, the mean RMR increases compared to baseline (P=0.165). Estimated daily calorie expenditure also increases.
After two weeks of supplementation with the supplement SeroVitalTM, both RMR and estimated daily calorie expenditure of the canine subjects tend to increase, evidencing the potential of the supplement to impart long-term fat burning effects. Additionally, endurance as measured by VO2max in the post-absorptive state significantly improves. Overall, the canine subjects administered with the SeroVitalTM supplement will show enhanced parameters of endurance, energy, and vitality.
Example 3
The effect of the nutritional supplement on lean body mass and weight change of canines is studied. The canine subjects in the first group are given daily doses of the nutritional supplement for 6 months at different dosing concentrations of active ingredients, while the canine subjects in the second group are given placebo. Weight, percent lean body mass, and percent fat body mass are chosen as primary measures of body composition at the 6 month analysis period. The weight, body composition, and physical performance (including endurance) of the canine subjects are determined before and after the treatment, along with hormonal data analysis.
The canine subjects treated with the nutritional supplement show a sustained increase in lean body mass and an improved physical performance.
While embodiments of the present invention have been described herein for purposes of illustration, many modifications and changes will become apparent to those skilled in the art. Accordingly, the appended claims are intended to encompass all such modifications and changes as fall within the true spirit and scope of this invention.

## Claims

1. A method of increasing growth hormone in a canine in need thereof, comprising:
providing a nutritional supplement, consisting essentially of:
about 1 mmol L-arginine, about 1 mmol Oxo-proline, about 2 mmol L-lysine, about 1.5 μmol N-acetyl L-cysteine, about 2 μmol L-glutamine, and about 125 μg Schizonepta (aerial parts) powder; and

administering an effective amount of the nutritional supplement to the canine.

2. The method of claim 1, wherein administering the nutritional supplement comprises orally administering.

3. The method of claim 1, wherein administering the nutritional supplement comprises administering about 2.9 grams of the nutritional supplement.

4. The method of claim 1, wherein the nutritional supplement is in powder, tablet, capsule, liquid, or wafer form.

5. The method of claim 1, wherein the nutritional supplement is administered once a week or once a month.

6. The method of claim 1, wherein the nutritional supplement is administered in an amount effective to reduce weight, increase endurance, increase fat metabolism, and/or increase lean body weight in the canine.

7. A method of increasing growth hormone in a canine, comprising:
providing a nutritional supplement, consisting essentially of:
0.86 mmol L-arginine;
1.32 mmol Oxo-proline:
2.05 mmol L-lysine;
1.53 μmol N-acetyl L-cysteine;
1.71 μmol L-glutamine; and
125 μg Schizonepta (aerial parts) powder; and

administering an effective amount of the nutritional supplement to the canine.

8. The method of claim 1, wherein administering the nutritional supplement comprises orally administering.

9. The method of claim 7, wherein administering the nutritional supplement comprises administering about 2.9 grams of the nutritional supplement.

10. The method of claim 7, wherein the nutritional supplement is in powder, tablet, capsule, liquid, or wafer form.

11. The method of claim 7, wherein the nutritional supplement is administered once a week or once a month.

12. The method of claim 7, wherein the nutritional supplement is administered in an amount effective to reduce weight, increase endurance, increase fat metabolism, and/or increase lean body weight in the canine.