IT202100028997A1 - USEFUL COMPOSITIONS TO REDUCE COGNITIVE DECLINE IN PETS - Google Patents

Description

Description of the patent for an industrial invention entitled:

?USEFUL COMPOSITIONS TO REDUCE COGNITIVE DECLINE IN PET ANIMALS?

The object of the invention is compositions comprising mangaferin or an extract containing it, bacosides or an extract containing them, Hericium erinaceus and luteolin or a plant source thereof. The compositions of the invention are useful for counteracting and preventing cognitive decline in pets, particularly dogs.

State of the art

Cognitive dysfunction in dogs? a neurodegenerative condition characterized by changes that affect cognitive decline and memory; The biological and behavioral characteristics of CCD (Canine Cognitive Dysfunction) are similar to what is observed in humans during the processes of aging and cognitive decline associated with age. in humans and in Alzheimer's disease ( 2017).

For example, in older Beagle breed dogs? A 30% loss of hippocampal neurons was observed followed by a consequent decrease in ability. cognitive ( 2008). Studies on other dog breeds have confirmed these findings (2008). In addition to the accumulation of amyloid beta plaques, recent studies have shown that the increase in the number of astrocytes and microglia correlates with the worsening of CCD ( 2016). In recent years, many researchers have recognized the importance of astrocytes and microglia in neuronal homeostasis and involvement in various pathologies. In humans? It has been demonstrated that the presence of populations of reactive microglia can be considered a biomarker of Alzheimer's disease and that these cells contribute to the mechanisms of neuronal damage and loss of neuronal capacity. cognitive ( 2012). Indeed, ? It has been demonstrated that inflammation has a central role in acute and chronic neurological disorders and? often dependent on the release of inflammatory mediators by these cells ( 2017). Neuro-inflammatory processes can be managed through the use of drugs, natural active ingredients, microbiota modulation and foods ( 2019, 2021).

In both humans with Alzheimer's disease and dogs with CCD, diet or the addition of supplements have a substantial impact on the development and progression of cognitive decline. In the dog? For example, it has been shown that foods rich in carnitine, alpha-lipoic acid, polyunsaturated fatty acids, carotenoids, vitamin C and E can cause a significant increase in physical capacity. cognitive ( 2003).

Together with diet therapy, numerous therapeutic proposals have been put forward for CCD with variable evidence both in terms of improving cognitive functions and slowing down the progression towards full-blown cognitive decline. These include the use of phytotherapeutics such as curcumin, resveratrol, tea catechins green, herbal extracts deriving from Chinese medicine including Ginkgo biloba, Huperzia serrata, Panax ginseng, Coptis chinensis. Recent literature on the prevention and treatment of cognitive decline in mouse models of Alzheimer's disease and the results obtained in humans reinforce the idea of a possible positive effect achievable through the use of standardized extracts and derivatives typical of Chinese medicine (

2019). Associations of phytotherapeutic extracts have been described as useful for combating memory loss and cognitive decline. US 20180369306 describes compositions including numerous components including Hericium erinaceus and Bacopa monnieri. EP 3 487 491 describes the association of cyperus extract with mangaferin or with an extract containing it useful for reducing stress and improving mood.

WO 2019038454 describes formulations including shilajit gum resin, Whitania (Ashwaganha) and possibly Bacopa monnieri and Hericium erinaceus.

EP 2785354 describes compositions including the flavonoid vicenin 2. Bacopa monnieri, Mangifera indica, Whitania somnifera are mentioned as possible additional components together with many other plants. US 2018/0021326 describes compositions based on psilocybin, psilocins, erinacins and other compounds extracted from mushrooms. Among the possible components mentioned are Bacopa monnieri, Cannabis, Hericium.

Description of the invention

Yes ? It has now been found that an association of mangaferin, bacosides, luteolin, in isolated form or of extracts or plant sources containing them, and Hericium erinaceus, determines advantageous and/or synergistic effects for the prevention and treatment of cognitive decline in livestock animals. company, particularly in dogs.

In its first aspect, the invention therefore has as its object compositions comprising mangaferin or an extract containing it, bacosides or an extract containing them, Hericium erinaceus and luteolin or a plant source thereof.

A particularly rich source of mangaferin, a derivative of xanthone present in plants belonging to the Anacardiaceae and Gentianaceae families? the mango (Mangifera indica). For the purposes of the invention, known extracts of seeds, fruits or bark of Mangifera indica can be used. Aqueous or hydroethanolic extracts, in particular of seeds, with mangaferin content equal to or greater than 90% by weight are preferred.

Bacosides are triterpene saponins contained in the aquatic plant Bacopa monnieri, used in Ayurvedic medicine to improve memory. Extracts from the aerial parts of Bacopa monnieri typically have a bacoside content of 20% to 50% by weight, depending on the extraction solvent (water, ethanol-water mixtures or other). The compositions of the invention can contain said extracts or bacosides in an isolated and purified form.

Even luteolin, a flavone present in many plants (thyme, sage, carrot, dandelion, peanut) can be present in the compositions of the invention in substantially pure form or in the form of an extract containing it. AND? preferred an extract of Arachis hypogaea fruit peels.

Hericium erinaceus, an edible basidiomycete used in Chinese medicine, can be in the form of a dried powder or an aqueous extract titrated in polysaccharides.

The compositions of the invention can also contain one or more components chosen from extracts of Oryza sativa (L.), Opuntia ficus indica (L.), Whitania somnifera (Ashwaganda), Passiflora edulis, Cannabis sativa. Cannabis sativa extract is preferably free from tetrahydrocannabinol and with a cannabidiol content varying from 0.1 to 20% by weight. A mixture of extracts of Oryza sativa (L.) and Opuntia ficus indica? commercially available (Actrisave?). An extract of Bacopa monnieri? present on the market as Bacopin?.

The quantities? per unit dose of the components will obviously depend on the species of animal and weight. Generally speaking, they will however be included in the following ranges:

- Bacopa monnieri extract from 1 to 300 mg, preferably from 1 to 25 mg; - Mangifera indica extract from 1 to 300 mg, preferably from 1 to 25 mg - Hericium erinaceus from 1 to 300 mg, preferably from 1 to 25 mg

- Luteolin from 1 to 50 mg.

If Mangiferin and bacosides are used in isolated form, the unit doses will correspond to those indicated above, taking into account the active ingredient content indicated for the extracts.

Can the optional components be present in quantity? from 1 to 200 mg.

The activity of the components of the compositions of the invention? was evaluated on an in vitro model (on murine cells) of neuronal inflammation mediated by microglial cells (Velagapudi et al. 2018). The compounds under examination have shown the ability to restore the up-regulation and down-regulation of markers of neuronal inflammation induced by activated microglia.

Hippocampal neurons of murine species, previously differentiated in specific culture medium, were put in contact, in co-culture for 24h, with soluble factors produced by microglial cells inflamed with 100 ng/ml LPS and IFN? 5 ng/ml for 24h. The genes most up-regulated and down-regulated by inflammation were then evaluated by transcriptomic analysis, with the aim of finding target genes of neuronal inflammation. Those that are significantly related to neurodegenerative diseases in the literature were then selected. In a second step, the inflamed cultures were treated with the selected extracts, which were then evaluated for their ability to to restore gene expression of target genes induced by inflammation.

The effect of the inflammatory stimulus in neurons after 24 hours of treatment of the following genes? been reversed as follows:

Hericium: Cxcl9 (0.333 ? 0.014; p<0.05); Gbp5 (0.339 ? 0.009; p<0.05); Ifi44 (0.176 ? 0.030; p<0.05); Usp18 (0.208 ? 0.030; p<0.05); Gbp8 (0.247 ? 0.061; p<0.05); Abcba1 (3.878 ? 0.110; p<0.01); Gpr56 (3.490 ? 0.168; p<0.05).

Bacopa: (Gbp5 0.313 ? 0.022; p<0.05); Ifi44 (0.123 ? 0.029; p<0.05); Usp18 (0.133 ? 0.019; p<0.05). Ashwaganda: Hp (1.954 ? 0.132; p<0.05); Serpine1 (3.826 ? 0.332; p<0.05).

Actrisave: Gbp5 (0.309 ? 0.006; p<0.05); Ifi44 (0.152 ? 0.039; p<0.05); Usp18 (0.175 ? 0.046; p<0.05); Gbp8 (0.251 ? 0.045 p<0.05); Abcba1 (2.662 ? 0.183 p<0.05).

Mangifera: Cxcl9 (0.411 ? 0.019; p<0.05); Ifi44 (0.159 ? 0.032 p<0.05); Usp18 (0.187? 0.031 p<0.05). Luteolin: Cxcl9 (0.645 ? 0.013; p<0.01); Gbp5 (0.511 ? 0.028; p<0.05); Ifi44 (0.334 ? 0.042 p<0.05); Gbp8 (0.566 ? 0.026 p<0.05)

Subsequent tests then showed synergistic effects (verified by obtaining results similar to those previously shown but also by reducing the concentrations of the products by 10 times) between Hericium, Bacopa, Ashwaganda, Mangifera, piceatannol (extracted from Passiflora edulis) , luteolin (extract from Arachis hypogaea) and cannabidiol (extract from Cannabis sativa).

The compositions of the invention may be formulated in a way suitable for oral administration and will be prepared according to conventional methods well known in pharmaceutical art, such as those described in "Remington's Pharmaceutical Handbook", using excipients, diluents, bulking agents, anti-caking agents , flavourings, sweeteners, acceptable for their end use.

The compositions of the invention will be presented in the form of capsules, tablets, wafers, granules, dispersible powders, suspensions, bars, biscuits. The compositions can also be used as additives to variously formulated foods or feeds (pet food) (jellies, kibbles, soups and the like).

In the formulations exemplified below, Hericium refers to a powder of sporophores of Hericium erinaceus, Bacopa refers to a hydroethanolic extract of Bacopa monnieri with a content of at least 20% bacosides determined by HPLC, Mangifera refers to an aqueous extract of Mangifera indica with a content of 90% of mangaferin determined by HPLC, Passiflora refers to a dry extract of Passiflora edulis seeds, Cannabis refers to an extract of Cannabis sativa free of THC and with a cannabidiol content from 0.1 to 20%, Ashwaganda yes refers to a dry extract of Whitania somnifera (whole plant) with a whitanolide content of 7% -10% by weight, Actrisave? ? a commercially available mixture of Oryza sativa (L.) and Opuntia ficus extracts.

Example 1 - Gastro-protected film tablets

Ingredient mg/tablet

Hericium 25

Bacopa 25

Mangifera 25

Luteolin 2

Cellulose 20

Silicon dioxide 20

Magnesium stearate 150

Gastro-resistant film 5

Other excipients to taste

TOTAL 500

Example 2 - Gastro-protected film tablets

Ingredient mg/tablet

Hericium 15

Bacopa 15

Mangifera 15

Luteolin 5

Actrisave? 10

Ashwaganda 10

Passionflower 10

Cannabis 10

Cellulose 20

Silicon dioxide 25

Magnesium stearate 100

Gastro-resistant film 5

Other excipients to taste

TOTAL 550

Example 3 - Immediate release tablets Ingredient mg/tablet Hericium 10 Bacopa 10 Mangifera 10 Luteolin 2.5 Sorbitol 90 Flavor 20 Magnesium stearate 7.5 Silicon dioxide 2.5 Acesulfame K 5.0 Croscarmellose sodium 6.0 Other excipients to taste Total 250

Example 4 - Slow-release tablets Ingredient mg/tablet Hericium 20 Bacopa 10 Mangifera 10 Luteolin 2.5 Actrisave? 15 Ashwaganda 15 Passion flower 15 Cannabis 5 Microcrystalline cellulose 90 Calcium phosphate 200 Hydroxypropyl methylcellulose 55 Magnesium stearate 5 Silicon dioxide 4 Other excipients to taste Total 600 Example 5 - Three-layer differentiated-release tablets Immediate-release internal layer

Ingredient mg/tablet Hericium 10 Bacopa 10 Mangifera 10 Sorbitol 20 Flavor 20 Magnesium stearate 2.5 Silicon dioxide 2.5 Acesulfame K 5.0 Croscarmellose sodium 6.0 Intermediate layer

Luteolin 5 Actrisave 10 Ashwaganda 10 Microcrystalline cellulose 100 Calcium phosphate 100 Magnesium stearate 4.5 Silicon dioxide 3.5 Croscarmellose sodium 1 Extended release layer

Ashwaganda 5 Passiflora 5 Cannabis 5 Calcium phosphate 120 Hydroxypropyl methylcellulose 100 Magnesium stearate 2 Silicon dioxide 2 Other excipients to taste Total 1500 Example 6 - Gold-dispersible formulation Ingredient mg/dose Hericium 30 Bacopa 10 Mangifera 15 Luteolin 1.5 Sorbitol 160 A rome orange 20 Mandarin flavor 5 Aerosil 5 Fructose 1250 Other excipients to taste Total 1500

Example 7 - Gummy discoid

Ingredient mg/dose Hericium 10 Bacopa 10 Mangifera 10 Luteolin 2 Actrisave 10 Ashwaganda 10 Cannabis 5 Gum base 200 Thyme aroma 2 Levilite 20 Talc F.U. 20 Vegetable magnesium stearate 15 Shellac 12 Xylitol 125 Gum arabic 6 Titanium dioxide 6 Carnauba wax 0.2 Other excipients to taste Total 500

Example 8? liquid form for oral use

Ingredient mg/dose Hericium 5 Bacopa 5 Mangifera 5 Luteolin 5 Actrisave 5 Ashwaganda 5 Cannabis 2 Magnesium gluconate 250 Zinc gluconate 25 Fructose 2500 Apricot flavor 25 Anhydrous citric acid 50 Coloring: Beta-carotene 1% 25 Aerosil 200 10 Water to taste

Example 9? Gel for topical application

Ingredient % Hericium 0.1 Bacopa 0.1 Mangifera 0.2 Luteolin 0.1 Actrisave 0.1 Ashwaganda 0.1 Cannabis 0.3 Sodium hyaluronate 0.5 Water to taste

Example 10? Dog biscuit

Ingredient % Spelled flour 19 Rye flour 27 Sunflower oil 3 Grated cheese 8 Oat flour 19 Hericium 0.1 Bacopa 0.1 Mangifera 0.2 Luteolin 0.1 Actrisave 0.1 Ashwaganda 0.1 Cannabis 0. 3 Water to taste at 100

Claims (11)

1. Compositions including mangaferin or an extract containing it, bacosides or an extract containing them, Hericium erinaceus and luteolin or a plant source thereof. 2. Compositions according to claim 1 in which the mangaferin source is? an extract of Mangifera indica and the source of bacosides? an extract of Bacopa monnieri. 3. Compositions according to claim 1 or 2 in which the source of luteolin is? an extract of Arachis hypogaea. 4. Compositions according to one of claims 1 to 3 further comprising one or more? components chosen from extracts of Oryza sativa (L.), Opuntia ficus indica (L.), Whitania somnifera, Passiflora edulis, Cannabis sativa. 5. Compositions according to one or more? of the previous claims in which the quantities? per unit dose of the components are included in the following ranges: - Bacopa monnieri extract from 1 to 300 mg; - Mangifera indica extract from 1 to 300 mg; - Hericium erinaceus from 1 to 300 mg; - luteolin from 1 to 50 mg. 6. Compositions according to one or more? of the previous claims in which the Bacopa monnieri extract is a hydroalcoholic extract with a bacoside content of at least 20% by weight. 7. Compositions according to one or more? of the previous claims in which the Mangifera extract indicates ? a hydroalcoholic extract with a mangaferin content of at least 90%. 8. Compositions according to one or more? of the previous claims in which Hericium erinaceus? in the form of dried powder or aqueous extract titrated in polysaccharides. 9. Compositions according to one or more? of the previous claims comprising extracts of Oryza sativa (L.), Opuntia ficus indica (L.), Whitania somnifera, Passiflora edulis and Cannabis sativa. 10. Compositions according to one or more? of the previous claims in the form of capsules, tablets, wafers, granules, dispersible powders, suspensions, bars, biscuits, gels for topical application. 11. Compositions of claims 1-10 for use in the treatment of cognitive decline in companion animals.