COMPOUND FOR TREATING OXIDATIVE STRESS IN MAMMALS

Abstract

Managing and treating elevated OS biomarkers in mammals such as companion animals with at least one of the supplements alpha-lipoic acid, carnitine, co-enzyme Q-10, ginger, green tea, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, vitamin E or selenium. Diagnosing an oxidative stress (OS) in a mammal comprises collecting a sample; screening the sample to detect the presence of an OS biomarker, selectively isoprostane and other antioxidants such as HODE microRNAs. TAC: GSH, MDA, and TNF-alpha. The sample can be saliva.

Claims

1-20. (canceled)

21. A veterinary nutritional compound for treating an elevated isoprostane OS biomarker in a dog with supplements of at least vitamin D and selenium, wherein the isoprostane biomarker in the dog so treated is reduced by at least 40% after five to six months from a level below 1.75 ng/ml as measured in saliva from the dog.

22. A veterinary nutritional compound as claimed in claim 21 including the supplements of at least alpha-lipoic acid, co-enzyme Q-10, green tea, turmeric.

23. A veterinary nutritional compound as claimed in claim 21 including treating an elevated HODE OS biomarker in the dog.

24. A veterinary nutritional compound as claimed in claim 22 including treating an elevated HODE OS biomarker in the dog.

25. A compound as claimed in claim 21 including at least one of supplements selected from the group consisting of alpha-lipoic acid, co-enzyme Q-10, green tea, turmeric.

26. A compound as claimed in claim 21 including at least two supplements selected from the group consisting of alpha-lipoic acid, co-enzyme Q-10, green tea, turmeric.

27. A compound as claimed in claim 21 including at least three supplements selected from the group consisting of alpha-lipoic acid, co-enzyme Q-10, green tea, turmeric.

28. A compound as claimed in claim 21 including at least two supplements selected from the group consisting of carnitine, ginger, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, and vitamin E.

29. A compound as claimed in claim 21 including at least three supplements selected from the group consisting of carnitine, ginger, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, and vitamin E.

30. A compound as claimed in claim 22 including at least two supplements selected from the group consisting of carnitine, ginger, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, and vitamin E.

31. A compound as claimed in claim 22 including at least three supplements selected from the group consisting of carnitine, ginger, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, and vitamin E.

Description

DETAILED DESCRIPTION

[0030] Different biomarkers are described and different abbreviations below are as follows.

[0031] IsoP, isoprostane(s); AOX, antioxidant capacity; CRP, C-reactive protein; GSH, glutathione; MDA, malonaldehyde; NO, nitric oxide; TAC, total antioxidant capacity; transcription factor Nrf-2 (Nuclear factor erythroid 2-related factor 2), TNF-alpha, tumor necrosis factor-alpha, and HODE, hydroxyoctadecadienoic acid.

[0032] The disclosure provides a straightforward, reliable assay for OS biomarkers and for antioxidant capacity of biological fluids.

[0033] The present disclosure relates to a test for levels of oxidized lipid biomarkers, selectively isoprostanes, HODE, microRNA and other biomarkers for animals, particularly companion animals and more particularly dogs, cats or horses using saliva that permits the rapid, accurate, non-invasive quantitative screening for biolipids, selectively in the animal.

[0034] The saliva-based test assay quantifies the isoprostane, HODE and microDNA levels in dog saliva to determine if the pet's body is undergoing harmful OS. OS creates reactive oxygen species (ROS) causing cells to undergo damage and release biomarker lipids and enzymes that lead to tissue inflammation, infections, periodontal disease, obesity and even cancers. However, free radicals themselves are so reactive and short-lived that direct measurement is not possible. Thus, isoprostane and HODE levels serve as reliable surrogate biomarkers for the presence of ROS.

[0035] The saliva-based tests of this disclosure are novel isoprostane, HODE and microRNA tests, and are examples of a set of unique biomarker tests for pets that can be measured in saliva.

[0036] For quantitative testing, an animal's serum or saliva or other bodily fluid is added to the ELISA microtiter plate or other immunoassay platforms such as but not limited to lateral flow, or latex or bead agglutination, which measures the presence of cellular oxidative stress.

[0037] Once collected at or received by the lab, the blood serum or saliva or other bodily fluid sample is then screened using the ELISA method or other immunoassay platforms such as but not limited to lateral flow, or latex or bead agglutination, which measures the presence of OS.

[0038] Forms of biological fluid, other than saliva, for instance urine, tears, sweat, or milk or other mucosal secretions can be used.

[0039] The detection of cellular oxidative stress can be performed with an immunoassay. Immunoassays include, but are not limited to, ELISA test, RIA test, latex agglutination, beads assay, and proteomic assays. A preferable immunoassay is the ELISA test. Other immunoassays can be used and the choice of immunoassay can be determined by one of ordinary skill in the art.

[0040] A method for diagnosing OS in a mammal such as a human, dog or other companion animal comprises the steps of collecting a sample of saliva; screening the sample to detect a level of at least one of a number of OS markers and detecting and diagnosing the presence of OS based on the level of one or more markers.

[0041] A method for diagnosing OS in a mammal such as human, dog or other companion animal comprises the steps of collecting a sample of saliva; screening the sample to detect a level of at least one isoprostane, and detecting and diagnosing the presence of OS based on the quantitative level of the isoprostane.

[0042] A method for diagnosing OS in a mammal such as human, dog or other companion animal comprises the steps of collecting a sample of saliva; screening the sample to detect a level of at least one HODE, and detecting and diagnosing the presence of OS based on the quantitative level of the HODE.

[0043] The method further comprises collecting a first testing portion of the saliva sample and wherein the first testing portion is the sample for use in the screening step. [Glucuronidase pretreatment is not required for saliva samples.]

[0044] The method includes the screening step utilizes an enzyme-linked immunosorbent assay (ELISA) testing system to detect the level of the saliva-based OS marker.

Results

[0045] After completing the initial clinical trial studies, analyzing 282 clinical patient samples; 79 of them (35%) were positive, having isoprostane biolipid levels, for example, above the normal reference range we have established (i. e., 0.5-1.75 ng/mL of saliva).

[0046] Of the 79 positive testing dogs, there were: 34% spayed females, 32% neutered males, 21% intact males and 15% intact females. The ages ranged from 4 months to 15 years, although most were middle aged or older. The weight range was 4-143 pounds, with 84% being medium to large or giant in size; no breed type predominated.

[0047] The diets fed the 79 positive dogs included: 40% ate only a commercial raw diet; 20% only a commercial dry kibble; 13% a home cooked or home prepared raw diet, and 3% ate a combination of a commercial kibble and raw.

[0048] Of the 79 positive dogs, 38 also had saliva-based profiles run for food sensitivity and intolerances to 24 foods. Interestingly, only 3 of these 38 dogs had Saliva based test results that were reactive (20 or more foods). These results suggest that dogs with clinical issues related to intense itching, scratching, chewing, and bowel irritability had relatively few identified foods as the culprits. Environmental exposure to inhalants, fleas, ticks, mites and other insects as well as contact reactants could be involved; and 3 dogs were taking an isooxazoline parasiticide.

[0049] Positive testing dogs should be retested in about 6 months after being on foods and supplements designed to lower their OS. The beneficial outcome of 50 isoprostane positive samples revealed reduced isoprostane levels when retested 5-6 months after taking the supplements listed below.

Management and Treatment

[0050] Management and treatments measures to alleviate elevated oxidative stress biomarkers of at least isoprostane and/or HODE, and their clinical expression in companion animals such as dogs or cats, and other animal species are described below. Use of supplements for isoprostane and HODE can be used for any other biomarkers of OS such as MicroRNAs PGF2a, MDA, TAC and others. Supplements that are used to bring down high levels of isoprostane and/or HODE are: [0051] Alpha-Lipoic acid [0052] Carnitine [0053] Co-Enzyme Q-10 [0054] Ginger [0055] Green tea [0056] Licorice [0057] Milk thistle, and a few more like garlic and honey. [0058] Resveratrol (as a natural supplement or as food like blueberries and cranberries) [0059] Selenium [0060] Soybeans [0061] Tomatoes [0062] Turmeric (curcumin)—without black pepper for pets [0063] Vitamin D [0064] Vitamin E

[0065] A synergistic biological effect of mutual benefit, based upon the principle of cooperative symbiosis, is achieved by this group of ingredients.

[0066] Two or more of Alpha-Lipoic acid; Carnitine, Co-Enzyme Q-10, Ginger, Green tea, Licorice, and Milk thistle produce enhanced and thus are synergistic.

[0067] The synergy results from the interaction between nutrients, their absorption and bioavailability in the body which in this situation is positive and provides health benefits. Evidence-based studies have illustrated that the active components in the nutritional supplements of the disclosed supplements work together synergistically to enhance their functional properties in preventing diseases and betterment of health. The role of the disclosed nutritional supplements working together enhances treatment and prevention of chronic diseases, such as cardiovascular and periodontal disease, infections, dysbiosis, diabetes, obesity, and cancers. This combination synergic effect is a scientific advance in nutritional therapy and significantly brings down high levels of isoprostane and/or HODE more effectively than when the supplements are separately used.

[0068] Additional synergistic effects can be obtained with Garlic and honey, Resveratrol (as a natural supplement or as food like blueberries and cranberries); Selenium; Soybeans; Tomatoes; Turmeric (curcumin); Vitamin D; and Vitamin E.

[0069] Each of these individually produce overtly similar effects, they display greatly enhanced effects when given in combination. The combined effect is greater than that predicted by their individual potencies, and thus the combination is synergistic.

[0070] The compound of the disclosure includes treating an elevated isoprostane OS biomarker with at least one of alpha-lipoic acid, co-enzyme Q-10, green tea, turmeric, vitamin D, or selenium.

[0071] The method of the disclosure includes treating an elevated isoprostane OS biomarker and an elevated HODE OS biomarker with at least one of alpha-lipoic acid, co-enzyme Q-10, green tea, turmeric, vitamin D, or selenium.

[0072] The compound of the disclosure includes treating an elevated HODE OS biomarker with at least one of alpha-lipoic acid, co-enzyme Q-10, green tea, turmeric, vitamin D, or selenium.

[0073] The compound of the disclosure includes treating an elevated OS biomarker being at least TAC, GSH, MDA, TNF-alpha, NO, and microRNA with at least one of alpha-lipoic acid, co-enzyme Q-10, green tea, turmeric, vitamin D, or selenium.

[0074] The compound of the disclosure includes treating an elevated isoprostane OS biomarker with at least alpha-lipoic acid, co-enzyme Q-10, green tea, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, or selenium.

[0075] The compound of the disclosure includes treating an elevated HODE OS biomarker with at least alpha-lipoic acid, co-enzyme Q-10, green tea, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, or selenium.

[0076] The compound of the disclosure includes treating an elevated isoprostane OS biomarker with at least one of, or with at least two of, or with at least three of, or with at least four of, or with at least five of, or with at least six of, or with at least seven of, or with at least eight of, or with at least nine of, or with at least ten of alpha-lipoic acid, carnitine, co-enzyme Q-10, ginger, green tea, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, vitamin E or selenium.

[0077] The compound of the disclosure includes treating an elevated isoprostane OS biomarker and an elevated HODE OS biomarker with at least one of, or with at least two of, or with at least three of, or with at least four of, or with at least five of, or with at least six of, or with at least seven of, or with at least eight of, or with at least nine of, or with at least ten of: alpha-lipoic acid, carnitine, co-enzyme Q-10, ginger, green tea, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, vitamin E or selenium.

[0078] The compound of the disclosure includes treating an elevated HODE OS biomarker with at least one of, or with at least two of, or with at least three of, or with at least four of, or with at least five of, or with at least six of, or with at least seven of, or with at least eight of, or with at least nine of, or with at least ten of alpha-lipoic acid, carnitine, co-enzyme Q-10, ginger, green tea, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, vitamin E or selenium.

[0079] The compound of the disclosure includes treating an elevated OS biomarker being at least TAC, GSH, MDA, TNF-alpha, NO, and microRNA with at least one of, or with at least two of, or with at least three of, or with at least four of, or with at least five of, or with at least six of, or with at least seven of, or with at least eight of, or with at least nine of, or with at least ten of alpha-lipoic acid, carnitine, co-enzyme Q-10, ginger, green tea, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, vitamin E or selenium.

[0080] The compound of the disclosure includes treating an elevated OS biomarker being at least TAC, GSH, MDA, TNF-alpha, NO, and microRNA with at least one of, or with at least two of, or with at least three of, or with at least four of, or with at least five of, or with at least six of, or with at least seven of, or with at least eight of, or with at least nine of, or with at least ten of: alpha-lipoic acid, carnitine, co-enzyme Q-10, ginger, green tea, licorice, milk thistle, garlic, honey. resveratrol, soybeans, tomatoes, turmeric, vitamin D, vitamin E or selenium.

[0081] The proportional decrease in the biomarker level of the oxidative stress, being the value of the isoprostane and/or HODE has been at least 40% and in some cases as much as 90% after about 5-6 months of ingestion of the compounds of the disclosure.

[0082] The compound is formulated with the ingredients being mixed together in powdered form and being for ingestion and digestion by the mammal, selectively an animal at least once or twice per day together with the food that is being eaten. In this format, the powder is mixed into the food prior to the food being given to the mammal such as human or animal.

General

[0083] Many different formats are possible. In the specification, there have been disclosed typical embodiments of the disclosure. 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 disclosure being set out in the claims. It is therefore to be understood that within the scope of the appended claims the disclosure may be practiced otherwise than as described.