GB2547903A

GB2547903A - Antioxidant mix for learning

Info

Publication number: GB2547903A
Application number: GB1603585.9A
Authority: GB
Inventors: Lowe John
Original assignee: Dodson & Horrell Ltd
Current assignee: Dodson & Horrell Ltd
Priority date: 2016-03-01
Filing date: 2016-03-01
Publication date: 2017-09-06
Also published as: GB201603585D0

Abstract

An animal food supplement for use in improving cognitive function in animals comprises one or more natural plant-based antioxidants, one or more sources of vitamin C and one or more sources of vitamin E. The plant-based antioxidants may comprise bioflavonoids such as polyphenolic flavonoids and may be present as a fruit, plant, fruit extract or plant extract, such as beetroot, spinach, pomegranate, blackcurrants or blackberries. The vitamin C source may comprise ascorbic acid and/or rosehip while the vitamin E source may comprise mixed tocopherols and/or tocotrienols. The supplement may also contain rosemaric acid and sacrificial antioxidants including gallic acid, a source of carotenoids and one or more of carrot, tomato or oregano. A method feeding animals, such as dogs, horses, rabbits, livestock or poultry, the supplement to improving learning ability, memory or cognitive function is disclosed. This may involve providing an animal with 0.2g to 20g or from 1g to 5g of the supplement per kilogram of body weight per day.

Description

Antioxidant mix for learning

The present invention is in the field of animal food supplements. More particularly, the present invention relates to the new use of an antioxidant blend which provides various advantages to the health and wellbeing of animals. GB2385768 discusses an animal food supplement that comprises a blend of antioxidants. It is reported in this document how the antioxidants interact synergistically in the body so as to increase the total antioxidant capacity (TAG) of animals, increase their plasma vitamin E levels, increase the resistance of low density lipoprotein to oxidation, and to increase protection of intestinal cells in the gut. The antioxidant blend is designed to be a component of the diet of cats, dogs, rabbits and horses.

Flavonoid antioxidants have recently been reported to modulate memory and learning in animals when included at varying amounts in the daily diet when fed over a range of periods from a single dose to 7 months continuous feeding, such as in “Rendeiro, C., Guerreiro, J.D.T., Williams, C.M., and J.P.E Spencer (2012) Flavonoids as modulators of memory and learning: molecular interactions resulting in behavioural effects. Proc. Nutr. Soc. 71 246-262”. This is believed in part to be due to their antioxidant capacity, but also through other yet unidentified actions in the animal, such as regulation of oxidative stress, enhancement of neuro-protective proteins and anti-inflammatory actions.

According to an aspect of the invention, there is provided an animal food supplement for use in improving cognitive function, learning, memory, or any combination thereof in animals, wherein the supplement comprises one or more naturally occurring plant-based antioxidants, one or more sources of vitamin C and one or more sources of vitamin E.

According to another aspect of the invention, there is provided the use of an animal feed supplement in the manufacture of a medicament for the improvement cognitive function, learning, memory, or any combination thereof in animals, wherein the animal feed supplement comprises one or more naturally occurring plant-based antioxidants, one or more sources of vitamin C and one or more sources of vitamin E.

According to another aspect of the invention, there is provided a method of improving cognitive function, learning, memory, or any combination thereof in animals, wherein the method comprises administering an animal feed supplement to an animal, wherein the supplement comprises one or more naturally occurring plant-based antioxidants, one or more sources of vitamin C and one or more sources of vitamin E.

Preferably, the at least one naturally occurring plant-based antioxidant comprises one or more bioflavonoids. Preferably, the one or more bioflavonoids comprise polyphenolic flavonoids. Preferably, the at least one naturally occurring plant-based antioxidant is present in the animal feed supplement in the form of a fruit extract or plant extract. However, the plant-based antioxidant may be present in the form of a plant perse or fruit perse.

Preferably, the supplement for use in the invention comprises at least two plant-based antioxidants, optionally such that the antioxidants function synergistically. GB2385768 discusses how a combination of naturally occurring plant based antioxidant components, vitamin C and vitamin E, is particularly effective in elevating total antioxidant capacity (TAG), plasma vitamin E levels and the resistance of low density lipoprotein oxidation, when administered over a suitable period of time. It is discussed how combinations of naturally occurring plant based antioxidant components and vitamins C and E, show synergism in their antioxidant effectiveness. Thus, whilst each vitamin and each antioxidant component is effective on its own to some degree in increasing the TAG of an animal, the supplements of GB2385768 are much more effective than the simple additive effect of the ingredients. Furthermore, the blend of antioxidants can cover all the phases in the animal (lipid, aqueous, membrane intra and intercellular etc) where antioxidant activity is needed, and they can thus deal with the variety of free radicals produced. Furthermore, the antioxidants in the blend support each other in a cascadelike fashion with a sacrificial antioxidant (eg. gallic acid) at the end of the chain. This is an extremely valuable effect and is believed to contribute substantially to the effectiveness of these supplements. Thus, what happens is that when any antioxidant reacts with a free radical, it in itself becomes oxidized. This first oxidized antioxidant is not very reactive, however, so is not especially dangerous. Because it is oxidized and is itself a free radical, it will react with another second antioxidant, providing the H-donating ability of the second antioxidant is chemically acceptable to the reactivity level of the oxidized first antioxidant. Thus, each antioxidant's ability to scavenge or donate H atoms relative to another is important.

Surprisingly, the inventors of the present invention have found that the blend of antioxidants discussed in GB2385768 can improve learning ability and cognitive function in animals, and particularly young animals. The blend of antioxidants has, surprisingly, been found to synergistically improve cognitive function and learning in animals. The supplement for use in the present invention has been found, surprisingly, to increase learning, memory and cognitive function in animals to a synergistic extent. That is, the supplement for use has been found to improve learning, cognitive function and memory to a greater extent than what would be expected by combining the total additive effect of each antioxidant component. Without being limited by theory, it is postulated that bioflavonoid antioxidants, which are known to increase learning, memory and cognitive function in animals, have this function synergistically improved by the presence of the vitamin C and Vitamin E antioxidants, and any other additional antioxidants. In a similar manner to as discussed in GB2385768, the antioxidants present in the animal food supplement for use in the present invention are believed to work better in teams than in isolation. For example, including two different, but complimentary antioxidants will have a greater impact than doubling either one.

The food supplement for use in the present invention is for use in enhancing the cognitive function, learning, memory, or any combination thereof of any animal, including humans. Preferably, the food supplement for use in the invention is for use in enhancing the cognitive function, learning, memory, or any combination thereof of dogs, cats, horses, rabbits, livestock or poultry. It is envisaged that the supplement for use in the invention will be able to be fed to pregnant and young animals where an enhancement in the animal’s memory, cognitive function and learning ability would mean the animal can be improved at carrying out a specific function or task. Examples of animals that would benefit from an enhancement in learning, memory and cognitive function include guide dogs and sheep dogs.

The food supplement for use in the invention can be used to improve learning, memory and/or cognitive function in animals of any age. Preferably, the food supplement is used to improve learning, memory and/or cognitive function in young animals. Preferably, this is accomplished by feeding the supplement to pregnant female animals or to their offspring from as soon as they are able to eat solid food until adult hood. Most preferably, this is accomplished by feeding the supplement to pregnant female animals and to their offspring from as soon as they are able to eat solid food until adulthood.

The supplement for use in the invention is preferably fed to the animals in an amount sufficient to improve their memory, learning ability and/or cognitive function. Typically, the supplement for use in the invention is fed to animals in an amount of from 0.1g to 30g per kilogram of the animal’s diet, and preferably from 0.2 g to 20 g per kilogram of the animal’s diet. Alternatively, the supplement for use in the invention is fed to the animal in an amount of from 0.2 g to 20 g per kilogram of bodyweight per day. Preferably, the supplement for use in the invention is fed to the animal in an amount of from 1 g to 5 g per kilogram of bodyweight per day. Alternatively, the supplement for use in the invention can be fed to the animals in an amount such that the animal is provided with from 250 mg to 750 mg of flavonoid per kg of bodyweight per day, and preferably from 350 mg to 500 mg of flavonoid per kg of bodyweight per day. Typically, the supplement for use in the invention is included in the diet of an animal in an amount of from 0.01 % to 5% by weight of the animal’s diet, and preferably in an amount of from 0.02 % to 2% by weight of the animal’s diet.

The animal supplement for use in the invention comprises one or more naturally occurring plant-based antioxidants, one or more sources of vitamin C and one or more sources of vitamin E. The one or more naturally occurring plant-based antioxidants typically comprise one or more bioflavonoids, preferably polyphenolic bioflavonoids. The one or more bioflavonoids may comprise flavanols such as quercetin (and derivatives); kaempferol; myricetin; galangin; flavanonols such as taxifolin and dihydrokaempferol; flavones such as luteolin (and derivatives); apigenin; chrysin; flavonones such as eriodictyol, hesperetin and naringenin (and derivatives); catechins; catechin gallates such as catechin, epicatechin, epigallocatechin, epicatechin gallate and epigallocatechin gallate; anthocyanidins such as cyanidin (and derivatives) and perlargonidin; and isoflavones such as genistein (and derivatives), biochanin, diadzein and formononetin; and any combination thereof.

The plant-based antioxidant components can be provided as parts of the plant (or fruit) per se, or as extracts therefrom. Normally, extracts are preferred. The extracts can contain other substances from the plants provided that they are not harmful in an animal feed. We have referred herein to the use of naturally occurring plant based antioxidants. We prefer to use plants or extracts (of the plants or of their fruits), especially since the other plant components present will normally be beneficial. However, synthetic equivalents can be used. As will be clear to those skilled in the art, the preferred plants are those containing high quantities of the desired antioxidants. Some plants are generally not very suitable for use in the present invention, one example being cereals.

Typical plants and plant extracts comprising one or more bioflavonoids suitable for use in the supplements for use in the invention comprise anthocyanin-containing Blueberry, Elderberry, grape skins and Bilberry plants, the extracts of which are particularly useful. Other typical plants, fruits and extracts thereof suitable for use comprise rutin; beetroot: spinach; pomegranate; red fruits and/or black fruits such as blackcurrants and blackberries; or any combination thereof. Preferably, the one or more bioflavonoids are present in the form of, or are extracted from rutin; beetroot; spinach; pomegranate; red fruits and/or black fruits such as blackcurrants and blackberries; or any combination thereof.

The plants, fruits, plant extracts or fruit extracts that comprise the one or more naturally occurring plant based antioxidants are typically present in the supplement for use in the invention in an amount of from 5% to 75% by weight of the supplement, and preferably, in an amount of from 5% to 25% by weight of the supplement.

Preferably, the supplement for use in the invention comprises from 150 g to 350 g of one or more flavonoids per kg of supplement, and more preferably from 200 g to 250 g of one or more flavonoids per kg of supplement.

The one or more sources of vitamin C for use in the supplement of the invention can be any suitable source of vitamin C. Typically, the source of vitamin C will be in a form (or protected) to avoid loss of essential function in processing and storage of the food supplements of the invention. The one or more sources of vitamin C for use in the supplement for use in the invention may typically comprise ascorbic acid. Vitamin C, Rosehip, or any combination thereof.

The one or more sources of vitamin C are typically present in the supplement for use in the invention in an amount of from 15% to 55% by weight of the supplement, and preferably in an amount of from 12% to 20% by weight of the supplement.

The one or more sources of vitamin E can comprise any suitable source of vitamin E. Typically, the one or more sources of vitamin E comprise mixed tocopherols, tocotrienols, or combinations thereof. Typically, the one or more sources of vitamin E are present in the supplement for use in the invention in an amount of from 5% to 30% by weight of the supplement, and preferably in an amount of from 5% to 15% by weight of the supplement.

The supplements for use in the present invention may contain additional antioxidants such as sacrificial antioxidants, and other additives that are considered beneficial to the health and/or wellbeing of the animal to which the supplement is to be administered. Additional antioxidants that can be included in supplements for use in the invention comprise gallic acid, taurine, glutathione, carotenoids, rosemaric acid, curcumin, tannin, bee pollen, parsley, oregano, thyme oil, tea polyphenols, or any combination thereof. Additional additives that can be included in supplements for use in the invention comprise oregano, carrot, tomato, gallic acid, rosemaric acid, and any combination thereof. Preferably, the supplements for use in the invention further comprise oregano, carrot, tomato, gallic acid and rosemaric acid.

Preferably, in the supplement for use in the invention, the one or more naturally occurring plant-based antioxidant is present in the form of rutin, beetroot, pomegranate, spinach, blackcurrant and blackberry; the one or more sources of vitamin C comprises ascorbic acid and rosehip; the one or more sources of vitamin E comprises mixed tocopherols; and the supplement further comprises rosemaric acid, gallic acid, tomato, carrot and oregano.

Most preferably, the supplement for use in the invention comprises the following ingredients in the following proportions.

Examples

Recent work in the Dodson and Horrell laboratories have shown the synergistic effects of the individual components of QLC, using slope ratio assay techniques. QLC is the commercially marketed name of the product disclosed in GB2385768. It comprises one or more naturally occurring plant-based antioxidant, one or more sources of vitamin C and one or more sources of vitamin E. The currently marketed formula of QLC, and the one tested in the experimental tests discussed below is as follows.

Slope ratio assays measure the relative effects of two treatments. The studies compared mixes of antioxidants with single antioxidants. For example, comparing rosehip with vitamin E indicates a value of 12.8:1, and comparing blackcurrant to vitamin E indicates a value of 3.35:1. One would thus predict a mix of equal amounts of the three to give 5.72:1 (calculated by determining the mean value of the antioxidants ((12.8+3.35+1)/3))), When measured, the value is 6.71:1, demonstrating the synergistic effect of mixed antioxidants over a single supply, and demonstrating that each antioxidant influences the activity of other antioxidants present in the supplement. This synergistic effect is the secret of the success and effectiveness of QLC; so it is much more beneficial than adding any one antioxidant to excess, which is often the case for example with vitamin E.

Slope ratio assay graph of a mix of three antioxidants compared to a standard vitamin E control.

Given the way in which QLC appears to work as a blend of antioxidants, the synergistic effect of one antioxidant supporting / protecting the other in the mix, it is believed that the delivery of the flavonoids in QLC along with additional antioxidants present in the QLC is sufficient to influence memory and learning in animals to a synergistic extent, and to a greater extent than if flavonoids were administered as the sole form of antioxidant in the supplement.

Tests on animals

In the study, a QLC composition with a formulation as detailed in the table above was included in the diet of rats at an amount to deliver 2g/kg body weight per day, equivalent to 432mg flavonoids / kg bodyweight per day. The experiment was carried out blind by the operatives.

The weight percentage of QLC in the diet ranged from 0.01-5 wt%.

Two groups of 20 rats per group were established at the start of the evaluation period, each group comprising 10 male and 10 female rats.

Group one (offspring from female rats fed QLC throughout pregnancy) received a diet containing QLC and group two (offspring from female rats not fed QLC during pregnancy) received an identical diet without QLC.

Suitable coding of the rats, cages and feed made it clear as to which group is which, but experimenters were not told which group is receiving the QLC feed. The rats were housed in single sex groups by treatment as appropriate using normal college facilities, husbandry practices and environmental conditions. Animals were routinely handled from as early as possible in life to accustom them to human contact.

The rats were fed the QLC diet free-choice until mature ~90 days of age during which time daily diet intake was recorded. 23 hours prior to the start of the training period food was withdrawn, to induce hunger to encourage learning via food reward in a maze. Subsequently rats were maintained at 0.85 of free feeding weight to maintain motivation in the study.

The rats from each group were randomly trained in a 12 choice T maze and then tested for learning ability at random in the same maze using a food reward. Measurements of the time taken to obtain the reward (T) and the number of erroneous turns (E) made in the maze were recorded, both during training and trial periods.

Training involved introducing the rat into the maze, initially near to the food reward, then gradually further from the reward. Initially, if the rat appears completely lost then some guidance was offered to help direct the rat to the food reward. Once the food had been found and consumed the rat was removed and allowed to rest before being repeated.

The maze was wiped down between rats to remove any odour traces that could confound the testing of subsequent rats.

During the trial period each rat was located at same start point, furthest from the feed, each time it is introduced into the maze. Testing was random by animal, each animal 3 tests per day over five days with a suitable, consistent (in terms of duration), rest-period between tests. An appropriate end to each trial period was when the rat gets to the reward or after a suitable period of time had elapsed.

The recorded results were plotted and analysed for significance using appropriate statistical techniques (cumulative and median latency plots and Cox regression analysis).

Results

Rats receiving QLC were significantly quicker through the maze than those not receiving QLC (P<0.05) at the start of the study and were also quicker at the end of the study duration. QLC fed rats made significantly less errors during the journey through the maze (P<0.05) Rats not fed QLC were significantly slower to commence learning the maze, started and finished with more incorrect turns.

The indications are that QLC as an antioxidant blend and supplier of flavonoids positively influences learning ability in young animals.

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Claims

Claims

1. An animal food supplement for use in improving cognitive function, learning, memory, or any combination thereof in animals, wherein the supplement comprises one or more naturally occurring plant-based antioxidants, one or more sources of vitamin C and one or more sources of vitamin E.
2. A supplement for use according to claim 1, wherein the one or more naturally occurring plant-based antioxidants comprises one or more bioflavonoids such as polyphenolic flavonoids.
3. A supplement for use according to claim 1 or 2, wherein the one or more naturally occurring plant-based antioxidants is present in the form of a fruit extract, plant extract, a fruit perse, a plant perse, or combinations thereof.
4. A supplement for use according to claim 3, wherein the fruit, plant, fruit extract or plant extract comprises rutin; beetroot; spinach; pomegranate; red fruits and/or black fruits such as blackcurrants and blackberries; or any combination thereof.
5. A supplement for use according to any preceding claim, wherein the source of vitamin C comprises ascorbic acid, rosehip, or a combination thereof.
6. A supplement for use according to any preceding claim, wherein source of vitamin E comprises mixed tocopherols, tocotrienols, or combinations thereof.
7. A supplement for use according to any preceding claim, wherein the supplement further comprises rosemaric acid.
8. A supplement for use according to any preceding claim, wherein the supplement further comprises one or more sacrificial antioxidants.
9. A supplement for use according to claim 8, wherein the one or more sacrificial antioxidants comprise gallic acid.
10. A supplement for use according to any preceding claim, wherein the supplement further comprises a source of carotenoids.
11. A supplement for use according to any preceding claims, wherein the supplement further comprises carrot, tomato, oregano, or any combination thereof.
12. A supplement for use according to any preceding claim, wherein the supplement comprises at least two plant-based antioxidants, optionally such that the antioxidants function synergistically.
13. A supplement for use according to any preceding claim, wherein the one or more naturally occurring plant-based antioxidant is present in the form of rutin, beetroot, pomegranate, spinach, blackcurrant and blackberry; wherein the one or more sources of vitamin C comprises ascorbic acid and rosehip; wherein the one or more sources of vitamin E comprises mixed tocopherols; and wherein the supplement further comprises rosemaric acid, gallic acid, tomato, carrot and oregano.
14. A supplement for use according to any preceding claim, wherein the supplement comprises
15. A supplement for use according to any preceding claim, wherein the use comprises improving the cognitive function, memory, learning ability, or any combination thereof of dogs, horses, rabbits, livestock and/or poultry.
16. A supplement for use according to any preceding claim, wherein the use comprises improving the cognitive function, memory, learning ability, or any combination thereof of animals prior to adulthood.
17. A supplement for use according to any preceding claim, wherein the use comprises improving the cognitive function, memory, learning ability, or any combination thereof of adult animals.
18. A supplement for use according to any preceding claim, wherein the use comprises providing the animal with the supplement in an amount of from 0.2 g to 20 g per kilogram of diet.
19. A supplement for use according to any preceding claim, wherein the use comprises providing the animal with the supplement in an amount of from 0.2 g to 20 g per kilogram of the animal’s body weight, per day, optionally, in an amount of from 1g to 5g per kilogram of the animal’s body weight, per day.
20. Use of an animal feed supplement in the manufacture of a medicament for the improvement of learning ability, memory, cognitive function, or any combination thereof in animals, wherein the animal feed supplement is as defined in any one of claims 1 to 14.
21. A method of improving learning ability, memory, cognitive function, or any combination thereof in animals, wherein the method comprises administering an animal feed supplement as defined in any of claims 1 to 14 to an animal.
22. Use according to claim 20, or a method according to claim 21, wherein the use or method of administration of the supplement is as defined in any one of claims 15 to 19.