WO2024200611A1 - Combination of trigonelline and oleuropein or oleuropein-metabolite for treating or preventing mitochondria-related conditions - Google Patents

Abstract

The compositions comprising trigonelline and at least one of oleuropein and/or metabolite thereof can be administered to a human to treat or prevent a mitochondria-related disease or a condition associated with altered mitochondrial function in an individual in need thereof or at risk thereof or in conditions of restrictions of NAD+ bioavailability. The mitochondria-related disease or condition is selected from the group consisting of deleterious effects of aging, stress (e.g., oxidative stress), obesity, overweight, reduced metabolic rate, metabolic syndrome, diabetes mellitus, complications from diabetes, hyperlipidemia, neurodegenerative disease, cognitive disorder, stress-induced or stress-related cognitive dysfunction, mood disorder, anxiety disorder, age-related neuronal death or dysfunction, chronic kidney disease, kidney failure, trauma, infection, cancer, hearing loss, macular degeneration, myopathies and dystrophies, and combinations thereof. The compositions can be administered to a human to promote the increase of intracellular levels of nicotinamide adenine dinucleotide ("NAD+") in cells and tissues for improving cell and tissue survival or overall cell and tissue health.

Description

TITLE

COMBINATION OF TRIGONELLINE AND OLEUROPEIN OR OLEUROPEIN-METABOLITE FOR TREATING OR PREVENTING MITOCHONDRIA-RELATED CONDITIONS

BACKGROUND

[001] The present disclosure generally relates to compositions and methods that use a combination of trigonelline and at least one oleuropein and/or metabolite thereof to produce intracellular NAD⁺/NADH and/or boost mitochondrial function. Intracellular levels of NAD+ can be increased in cells and tissues to improve cell and tissue survival and/or or overall cell and tissue health.

[002] Nicotinic acid and nicotinamide are the vitamin forms of nicotinamide adenine dinucleotide (NAD⁺). Eukaryotes can synthesize NAD⁺ de novo via the kynurenine pathway from tryptophan, and niacin supplementation prevents the pellagra that can occur in populations with a tryptophan-poor diet. Nicotinic acid is phosphoribosylated to nicotinic acid mononucleotide (NaMN), which is then adenylylated to form nicotinic acid adenine dinucleotide (NaAD), which in turn is amidated to form NAD⁺.

[003] NAD⁺ is an enzyme co-factor that is essential for the function of several enzymes related to reduction-oxidation reactions and energy metabolism. NAD⁺ functions as an electron carrier in cell metabolism of amino acids, fatty acids, and carbohydrates. NAD⁺ serves as an activator and substrate for sirtuins, a family of protein deacetylases that have been implicated in metabolic function and extended lifespan in lower organisms. The co-enzymatic activity of NAD⁺, together with the tight regulation of its biosynthesis and bioavailability, makes it an important metabolic monitoring system that is clearly involved in the aging process.

[004] Moreover, mitochondria are the primary source of aerobic energy production in mammalian cells and also maintain a large Ca²⁺ gradient across their inner membrane, providing a signaling potential for this molecule. Furthermore, mitochondrial Ca²⁺ plays a role in the mitochondria in the regulation of ATP generation and potentially contributes to the orchestration of cellular metabolic homeostasis. Specifically, activation of mitochondrial Ca2+ import increases cellular energy metabolism (Glancy & Balaban, 2012). SUMMARY

[005] In an embodiment, the present disclosure provides a method of treating or preventing (e.g., reducing incidence and/or severity) a mitochondria-related disease or a condition associated with altered mitochondrial function in a human in need thereof or at risk thereof. The method comprises orally administering to the human a composition comprising trigonelline and at least one of oleuropein and/or metabolite thereof.

[006] The mitochondria-related disease or condition can be selected from the group consisting of deleterious effects of aging, stress (e.g., oxidative stress), obesity, overweight, reduced metabolic rate, metabolic syndrome, diabetes mellitus, complications from diabetes, hyperlipidemia, neurodegenerative disease, cognitive disorder, stress-induced or stress-related cognitive dysfunction, mood disorder, anxiety disorder, age-related neuronal death or dysfunction, chronic kidney disease, kidney failure, trauma, infection, cancer, hearing loss, macular degeneration, myopathies and dystrophies, and combinations thereof.

[007] In another embodiment, the present invention provides a method of achieving at least one result selected from the group consisting of (i) improved mitochondrial calcium uptake in one or more cells, (ii) improved utilization of calcium in one or more cells, (iii) increased mitochondrial energy in one or more cells, (ii) improvement in a physiological state linked to metabolic fatigue in one or more cells, (iii) treatment or prevention of metabolic fatigue in one or more cells, (iv) treatment or prevention of muscle fatigue, (v) improved mobility, (vi) improved longevity (viii) increasing NAD+ levels in one or more cells and (ix) improved conditions of restrictions of NAD+ bioavailability, the method comprising orally administering to a human an effective amount of a combination of trigonelline and at least one of oleuropein and/or metabolite thereof..

[008] In a preferred embodiment, the trigonelline and at least one of oleuropein and/or metabolite thereof, is in an amount effective to increase the calcium uptake in mitochondrial cells.

[009] Moreover, the present combination of ingredient is also particularly effective in conditions of restrictions of NAD⁺ bioavailability. That condition has been reported in cancer and neurodegeneration, as well as the course of natural aging and, in general, during states of genotoxic stress, that accompany a growing list of diseases. In addition, loss of NAD homeostasis leads to progressive and reversible degeneration of skeletal muscle.

[0010] The increase in NAD⁺ biosynthesis can provide one or more benefits to the individual (e.g., a human undergoing medical treatment). The one or more benefits can comprise at least one of increased mitochondrial energy, treatment or prevention of metabolic fatigue, treatment or prevention of muscle fatigue, improvement in a physiological state linked to metabolic fatigue in one or more cells, improved mobility or improved longevity. Preferably, the NAD⁺ biosynthesis is increased in one or more cells of the human, for example one or more cells that are part of at least one body part selected from the group consisting of a liver, a kidney, a brain, and a skeletal muscle.

[0011] In some embodiments, at least a portion of the trigonelline is provided by a plant extract in the composition, such as one or more of a coffee extract, a hemp extract, a pumpkin seed extract and/or a fenugreek seed extract, for example a plant extract enriched in trigonelline.

[0012] In a preferred embodiment, at least a portion of trigonelline is provided from a fenugreek extract.

[0013] In some embodiments, at least a portion of the trigonelline is provided from an algae source, for example, a Laminariaceae extract.

[0014] In an embodiment, the metabolite of oleuropein is selected from the group consisting of oleuropein aglycone, hydroxytyrosol, homovanillyl alcohol, isohomovanillyl alcohol, glucuronidated forms thereof, sulfated forms thereof, derivatives thereof, and mixtures thereof.

[0015] In an embodiment, the composition is selected from the group consisting of a food product, beverage product, a food supplement, an oral nutritional supplement (ONS), a medical food, powdered nutritional products to be reconstituted in water or milk before consumption and combinations thereof.

[0016] An advantage of one or more embodiments provided by the present disclosure is to potentiate benefits on oxidative metabolism and prevent DNA damage.

[0017] Another advantage of one or more embodiments provided by the present disclosure is to replenish NAD⁺ pools, which decline with age.

[0018] Yet another advantage of one or more embodiments provided by the present disclosure is to help off-set slowing of the metabolism associated with aging.

[0019] Another advantage of one or more embodiments provided by the present disclosure is to help increase fatty acids metabolism.

[0020] Yet another advantage of one or more embodiments provided by the present disclosure is to help the body to metabolize fat and increase lean body mass.

[0021] Another advantage of one or more embodiments provided by the present disclosure is to help maintain heart health.

[0022] Yet another advantage of one or more embodiments provided by the present disclosure is to help support healthy LDL-cholesterol and fatty acid levels in the blood.

[0023] Another advantage of one or more embodiments provided by the present disclosure is to treat or prevent neurodegeneration, such as age-related neurodegeneration, and/or promote neurite outgrowth.

[0024] Additional features and advantages are described in, and will be apparent from, the following Detailed Description and the Figures.

BRIEF DESCRIPTION OF DRAWINGS

[0025] FIG. 1 is a graph showing the beneficial effect of the combination of 3pM Oleuropein aglycone plus 500pM Trigonelline iodine on mitochondria activation, via mitochondrial Ca²⁺, in C2C12-derived myotubes. The graph shows that Oleuropein aglycone alone (3pM) slightly enhances mitochondrial Ca²⁺ by 17%, compared with buffer (white). Conversely, the combination of Oleuropein aglycone (3pM) plus Trigonelline (500 pM) enhances mitochondrial Ca²⁺ by 76%, compared with Trigonelline alone (buffer, black). The data are plotted as a percentage of the integrated mitochondrial calcium response, obtained in presence of the buffer. Mitochondrial Ca²⁺ was stimulated in all the conditions with 5 mM caffeine. Results are expressed as mean +/- SEM, from n = 16 biological replicates, from 2 independent experiments. * indicates statistically significant difference between the conditions, at P < 0.05 (unpaired t-test).

[0026] FIG. 2 is a graph showing the beneficial effect of the combination of lOpM Oleuropein aglycone plus 500pM Trigonelline iodine on mitochondria activation, via mitochondrial Ca²⁺, in C2C12-derived myotubes. The graph shows that Oleuropein aglycone alone (lOpM) slightly enhance mitochondrial Ca²⁺ by 28%, compared with buffer (white). Conversely, the combination of Oleuropein aglycone (lOpM) plus Trigonelline (500 pM) enhances mitochondrial Ca²⁺ by 67%, compared with Trigonelline alone (buffer black). The data are plotted as a percentage of integrated mitochondrial calcium response obtained in presence of the buffer. Mitochondrial Ca²⁺ was stimulated in all the conditions with 5 mM caffeine. Results are expressed as mean +/- SEM, from n = 16 biological replicates, from 2 independent experiments. * indicates statistically significant difference between the conditions, at P<0.05 (unpaired test). DETAILED DESCRIPTION

[0027] Definitions

[0028] Some definitions are provided hereafter. Nevertheless, definitions may be located in the “Embodiments” section below, and the above header “Definitions” does not mean that such disclosures in the “Embodiments” section are not definitions.

[0029] All percentages expressed herein are by weight of the total weight of the composition unless expressed otherwise. As used herein, “about,” “approximately” and “substantially” are understood to refer to numbers in a range of numerals, for example the range of -10% to +10% of the referenced number, preferably -5% to +5% of the referenced number, more preferably -1% to +1% of the referenced number, most preferably -0.1% to +0.1% of the referenced number.

[0030] All numerical ranges herein should be understood to include all integers, whole or fractions, within the range. Moreover, these numerical ranges should be construed as providing support for a claim directed to any number or subset of numbers in that range. For example, a disclosure of from 1 to 10 should be construed as supporting a range of from 1 to 8, from 3 to 7, from 1 to 9, from 3.6 to 4.6, from 3.5 to 9.9, and so forth.

[0031] As used in this disclosure and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a component” or “the component” includes two or more components.

[0032] The words “comprise,” “comprises” and “comprising” are to be interpreted inclusively rather than exclusively. Likewise, the terms “include,” “including” and “or” should all be construed to be inclusive, unless such a construction is clearly prohibited from the context. Nevertheless, the compositions disclosed herein may lack any element that is not specifically disclosed herein. Thus, a disclosure of an embodiment using the term “comprising” includes a disclosure of embodiments “consisting essentially of’ and “consisting of’ the components identified. Any embodiment disclosed herein can be combined with any other embodiment disclosed herein.

[0033] The term “and/or” used in the context of “X and/or Y” should be interpreted as “X,” or “Y,” or “X and Y.” Similarly, “at least one of X or Y” should be interpreted as “X,” or “Y,” or “X and Y.”

[0034] Where used herein, the terms “example” and “such as,” particularly when followed by a listing of terms, are merely exemplary and illustrative and should not be deemed to be exclusive or comprehensive. As used herein, a condition “associated with” or “linked with” another condition means the conditions occur concurrently, preferably means that the conditions are caused by the same underlying condition, and most preferably means that one of the identified conditions is caused by the other identified condition.

[0035] The terms “food,” “food product” and “food composition” mean a product or composition that is intended for ingestion by an individual such as a human and provides at least one nutrient to the individual. A food product typically includes at least one of a protein, a lipid, a carbohydrate and optionally includes one or more vitamins and minerals. The compositions of the present disclosure, including the many embodiments described herein, can comprise, consist of, or consist essentially of the elements disclosed herein, as well as any additional or optional ingredients, components, or elements described herein or otherwise useful in a diet.

[0036] As used herein, the term "isolated" means removed from one or more other compounds or components with which the compound may otherwise be found, for example as found in nature. For example, “isolated” preferably means that the identified compound is separated from at least a portion of the cellular material with which it is typically found in nature. In an embodiment, an isolated compound is free from any other compound.

[0037] ‘ ‘Prevention” includes reduction of risk, incidence and/or severity of a condition or disorder. The terms “treatment,” “treat” and “to alleviate” include both prophylactic or preventive treatment (that prevent and/or slow the development of a targeted pathologic condition or disorder) and curative, therapeutic or disease-modifying treatment, including therapeutic measures that cure, slow down, lessen symptoms of, and/or halt progression of a diagnosed pathologic condition or disorder; and treatment of patients at risk of contracting a disease or suspected to have contracted a disease, as well as patients who are ill or have been diagnosed as suffering from a disease or medical condition. The term does not necessarily imply that a subject is treated until total recovery. The terms “treatment” and “treat” also refer to the maintenance and/or promotion of health in an individual not suffering from a disease but who may be susceptible to the development of an unhealthy condition. The terms “treatment,” “treat” and “to alleviate” are also intended to include the potentiation or otherwise enhancement of one or more primary prophylactic or therapeutic measure. The terms “treatment,” “treat” and “to alleviate” are further intended to include the dietary management of a disease or condition or the dietary management for prophylaxis or prevention a disease or condition. A treatment can be patient- or doctor-related.

[0038] The term "unit dosage form," as used herein, refers to physically discrete units suitable as unitary dosages for human subjects, each unit containing a predetermined quantity of the composition disclosed herein in an amount sufficient to produce the desired effect, in association with a pharmaceutically acceptable diluent, carrier or vehicle. The specifications for the unit dosage form depend on the particular compounds employed, the effect to be achieved, and the pharmacodynamics associated with each compound in the host.

[0039] As used herein, an “effective amount” is an amount that prevents a deficiency, treats a disease or medical condition in an individual, or, more generally, reduces symptoms, manages progression of the disease, or provides a nutritional, physiological, or medical benefit to the individual. The relative terms “improve,” “increase,” “enhance,” “promote” and the like refer to the effects of the composition disclosed herein, namely a composition comprising trigonelline, relative to a composition not having trigonelline but otherwise identical. As used herein, "promoting" refers to enhancing or inducing relative to the level before administration of the composition disclosed herein.

[0040] A "subject" or “individual” is a human. The term “elderly” means an age from birth of at least 60 years, preferably above 63 years, more preferably above 65 years, and most preferably above 70 years. The term “older adult” means an age from birth of at least 45 years, preferably above 50 years, more preferably above 55 years, and includes elderly individuals.

[0041] “Overweight” is defined for a human as a body mass index (BMI) between 25 and 30 kg/m². “Obese” is defined for a human as a BMI of at least 30 kg/m², for example 30-39.9 kg/m². “Weight loss” is a reduction of the total body weight. Weight loss may, for example, refer to the loss of total body mass in an effort to improve one or more of health, fitness or appearance.

[0042] " Diabetes" encompasses both the type I and type II forms of the disease. Nonlimiting examples of risk factors for diabetes include: waistline of more than 40 inches for men or 35 inches for women, blood pressure of 130/85 mmHg or higher, triglycerides above 150 mg/dl, fasting blood glucose greater than 100 mg/dl or high-density lipoprotein of less than 40 mg/dl in men or 50 mg/dl in women.

[0043] As used herein, the term "metabolic syndrome" refers to a combination of medical disorders that, when occurring together, increase the risk of developing cardiovascular disease and diabetes. It affects one in five people in the United States and prevalence increases with age. Some studies have shown the prevalence in the United States to be an estimated 25% of the population. In accordance with the International Diabetes Foundation consensus worldwide definition (2006), metabolic syndrome is central obesity plus any two of the following:

[0044] Raised triglycerides: > 150 mg/dL (1.7 mmol/L), or specific treatment for this lipid abnormality;

[0045] Reduced HDL cholesterol: < 40 mg/dL (1.03 mmol/L) in males, < 50 mg/dL (1.29 mmol/L) in females, or specific treatment for this lipid abnormality;

[0046] Raised blood pressure: systolic BP > 130 or diastolic BP >85 mm Hg, or treatment of previously diagnosed hypertension; and

[0047] Raised fasting plasma glucose: (FPG) > 100 mg/dL (5.6 mmol/L), or previously diagnosed type 2 diabetes.

[0048] As used herein, "neurodegenerative disease" or "neurodegenerative disorder" refers to any condition involving progressive loss of functional neurons in the central nervous system. In an embodiment, the neurodegenerative disease is associated with age-related cell death. Non-limiting examples of neurodegenerative diseases include mild cognitive impairment, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (also known as ALS and as Lou Gehrig's disease), peripheral neuropathy, AIDS dementia complex, adrenoleukodystrophy, Alexander disease, Alper's disease, ataxia telangiectasia, Batten disease, bovine spongiform encephalopathy (BSE), Canavan disease, corticobasal degeneration, Creutzfeldt-Jakob disease, dementia with Lewy bodies, fatal familial insomnia, frontotemporal lobar degeneration, Kennedy's disease, Krabbe disease, Lyme disease, Machado-Joseph disease, multiple sclerosis, multiple system atrophy, neuroacanthocytosis, Niemann-Pick disease, Pick's disease, primary lateral sclerosis, progressive supranuclear palsy, Refsum disease, Sandhoff disease, diffuse myelinoclastic sclerosis, spinocerebellar ataxia, subacute combined degeneration of spinal cord, tabes dorsalis, Tay-Sachs disease, toxic encephalopathy, transmissible spongiform encephalopathy, and wobbly hedgehog syndrome. The present disclosure is not limited to a specific embodiment of the neurodegenerative disease, and the neurodegenerative disease can be any neurologically-related condition known to one skilled in this art.

[0049] As used herein, "cognitive function" refers to any mental process that involves symbolic operations, e.g., perception, memory, attention, speech comprehension, speech generation, reading comprehension, creation of imagery, learning, and reasoning, preferably at least memory.

[0050] Methods for measuring cognitive function are well-known and can include, for example, individual or battery tests for any aspect of cognitive function. One such test is the Prudhoe Cognitive Function Test by Margallo-Lana et al. (2003) J. Intellect. Disability Res. 47:488-492. Another such test is the Mini Mental State Exam (MMSE), which is designed to assess orientation to time and place, registration, attention and calculation, recall, language use and comprehension, repetition, and complex commands. Folstein et al. (1975) J. Psych. Res. 12:189-198. Such tests can be used to assess cognitive function in an objective manner, so that changes in cognitive function, for example in response to treatment in accordance with methods disclosed herein, can be measured and compared.

[0051] As used herein, a “cognitive disorder” refers to any condition that impairs cognitive function. Non-limiting examples of a cognitive disorder include delirium, dementia, learning disorder, attention deficit disorder (ADD), and attention deficit hyperactivity disorder (ADHD).

[0052] As used herein, a "neurite" refers to any projection from the cell body of a neuron, such as an axon or a dendrite. The term is frequently used when speaking of immature or developing neurons, especially of cells in culture, because axons can be difficult to differentiate from dendrites before differentiation is complete. Neurites are often packed with microtubule bundles, the growth of which is stimulated by nerve growth factor (NGF), as well as tau proteins, microtubule associated protein 1 (MAPI), and microtubule associated protein 2 (MAP2). The neural cell adhesion molecule N-CAM simultaneously combines with another N-CAM and a fibroblast growth factor receptor to stimulate the tyrosine kinase activity of that receptor to induce the growth of neurites.

[0053] “Mobility” is the ability to move independently and safely from one place to another.

[0054] ‘ ‘Metabolic fatigue” means reduced mitochondrial function in one or more cells (e.g., one or more of liver, kidney, brain, skeletal muscle) due to a shortage of substrates within the one or more cells and/or due to an accumulation of metabolites within the one or more cells which interfere with mitochondrial function.

[0055] As used herein, “trigonelline” is any compound comprising 1-methylpyridin-l- ium-3 -carboxylate including, for example, any salt thereof (e.g., Chloride or Iodide salt) and/or a form in which the ring therein may be reduced.

[0056] In some embodiments, trigonelline is represented by the structure of formula 1, being able to establish a salt with an anion (X-), such as a halogen, for example, iodide or chloride. The structure of formula 1 is also known as 3 -carboxy- 1 -methylpyridinium, N-Methylnicotinic acid, l-methylpyridine-3 -carboxylic acid, 1-methylpyridin-l -ium-3 -carboxylic acid, Pyridinium 3 -carboxy- 1 -methyl- hydroxide inner salt (8CI), 1 -methylnicotinic acid, Pyridinium 3-carboxy-l - methyl-.

1

[0057] In some embodiments, trigonelline is represented by the structure of formula 2 in its inner salt form. The structure of formula 2 is also known as Caffearine, Gynesine, N-Methylnicotinate, Trigenolline, Coffearine, Trigonellin, Coffearin, Betain nicotinate, Betaine nicotinate, 1- methylpyridinium-3-carboxylate, Nicotinic acid N-methylbetaine, l-Methylpyridinio-3-carboxylate, 1- Methyl-3-pyridiniumcarboxylate, N-Methylnicotinic acid, Trigenelline, Caffearin, 3-Carboxy-l- methylpyridinium hydroxide inner salt, N'-Methylnicotinate, l-methylpyridin-l-ium-3-carboxylate, 3- Carboxy-1 -methylpyridinium hydroxide inner salt, Pyridinium 3-carboxy-l -methyl- hydroxide inner salt, l-methylpyridine-3 -carboxylic acid, l-methylpyridin-l-ium-3-carboxylic acid, 1 -methylnicotinate, Trigonelline (S), N-methyl-nicotinate, Pyridinium 3-carboxy-l -methyl- hydroxide inner salt (8CI), N'- Methylnicotinic acid, N-Methylnicotinic acid betaine, Nicotinic acid N- methylbetaine, 1 -Methyl- Nicotinic Acid Anion, Pyridinium 3-carboxy-l -methyl- inner salt, l-Methyl-5- (oxylatocarbonyl)pyridinium-3-ide, Pyridinium 3-carboxy-l -methyl- inner salt, 3-carboxy-l-methyl- Pyridinium hydroxide inner salt.

2

[0058] In some embodiments, optionally “trigonelline” can include metabolites and pyrolysis products thereof, such as nicotinamide, nicotinamide riboside, 1 -methylnicotinamide, l-methyl-2- pyridone-5-carboxamide (Me2PY), l-methyl-4-pyridone-5 -carboxamide (Me4PY), and alkylpyridiniums, such as 1-methyl-pyridinium (NMP) and 1,4-dimethylpyridinium; although as noted later herein, some embodiments exclude one or more of these metabolites and pyrolysis products of trigonelline.

[0059] Embodiments

[0060] The present disclosure provides a method for increasing intracellular NAD+ in a human, the method comprising administering to the human a composition comprising trigonelline and at least one of oleuropein and/or metabolite thereof in an amount effective to increase NAD⁺ biosynthesis.

[0061] In another embodiment, the present disclosure provides a method of treating or preventing (e.g., reducing incidence and/or severity) a mitochondria-related disease or a condition associated with altered mitochondrial function in a human in need thereof or at risk thereof. The method comprises orally administering to the human a composition comprising trigonelline and at least one of oleuropein and/or metabolite thereof in an amount effective to increase NAD⁺ biosynthesis.

[0062] The mitochondria-related disease or condition can be selected from the group consisting of deleterious effects of aging, stress, obesity, overweight, reduced metabolic rate, metabolic syndrome, diabetes mellitus, complications from diabetes, hyperlipidemia, neurodegenerative disease, cognitive disorder, stress-induced or stress-related cognitive dysfunction, mood disorder, anxiety disorder, age-related neuronal death or dysfunction, chronic kidney disease, kidney failure, trauma, infection, cancer, hearing loss, macular degeneration, myopathies and dystrophies, and combinations thereof.

[0063] The increase in NAD⁺ biosynthesis can provide one or more benefits to the individual (e.g., a human undergoing medical treatment). The one or more benefits can comprise at least one of increased mitochondrial energy, treatment or prevention of metabolic fatigue, treatment or prevention of muscle fatigue, improvement in a physiological state linked to metabolic fatigue in one or more cells, improved mobility or improved longevity. Preferably, the NAD⁺ biosynthesis is increased in one or more cells of the human, for example one or more cells that are part of at least one body part selected from the group consisting of a liver, a kidney, a brain, and a skeletal muscle. In some embodiments, the composition is administered to an older adult or an elderly individual.

[0064] The composition can comprise a pharmacologically effective amount of trigonelline in a pharmaceutically suitable carrier. In aqueous liquid compositions, the trigonelline concentration preferably ranges from about 0.05 wt.% to about 4 wt.%, or from about 0.5 wt.% to about 2 wt.% or from about 1.0 wt.% to about 1.5 wt.% of the aqueous liquid composition.

[0065] The method can comprise administering daily trigonelline in the weight range of 0.05 mg - 1 g per kg body weight, preferably 1 mg -200 mg per kg body weight, more preferably 5 mg - 150 mg per kg body weight, even more preferably 5 mg - 80 mg per kg body weight, or most preferably 5 mg - 20 mg per kg body weight.

[0066] Some embodiments comprise administering an amount of the composition that provides 1.0 mg to 10.0 g of the trigonelline / kg of body weight of the human, preferably 10 mg to 5.0 g of the trigonelline / kg of body weight of the human, more preferably 50 mg to 2.0 g of the trigonelline / kg of body weight of the human, most preferably 100 mg to 1.0 g of the trigonelline / kg of body weight of the human.

[0067] In some embodiments, at least a portion of the trigonelline is isolated. Additionally or alternatively, at least a portion of trigonelline can be chemically synthesized.

[0068] In one embodiment, the composition comprises trigonelline which is chemically synthesized which is at least about 90% trigonelline, preferably at least about 98% trigonelline.

[0069] In a preferred embodiment, at least a portion of the trigonelline is provided by a plant or algae extract, for example an extract from one or more of coffee bean (e.g., a green coffee extract), Japanese radish, fenugreek seed, garden pea, hemp seed, oats, potato, dahlia, Stachys species, Strophanthus species, Laminariaceae species (especially Laminaria and Saccharina), Postelsia palmaeformis, Pseudochorda nagaii, Akkesiphycus or Dichapetalum cymosum. The plant extract is preferably enriched in trigonelline, i.e., the starting plant material comprises one or more other compounds in addition to the trigonelline, and the enriched plant material has a ratio of the trigonelline relative to at least one of the one or more other compounds that is higher than the ratio in the starting plant material.

[0070] Therefore, some embodiments of the composition comprise plant sources and/or enriched plant sources that provide at least a portion of the trigonelline in the composition.

[0071] In a preferred embodiment, the composition comprises enriched fenugreek extract which provides at least about 25 - 50% trigonelline in the composition. In a more preferred embodiment, the composition comprises enriched fenugreek extract which provides at least about 28 - 40% trigonelline.

[0072] As used herein, a “composition consisting essentially of trigonelline” contains trigonelline and is substantially free or completely free of any additional compound that affects NAD⁺ production other than the trigonelline. In a particular non-limiting embodiment, the composition consists of the trigonelline and one or more excipients.

[0073] As used herein, “substantially free” means that any of the other compound present in the composition is no greater than 1.0 wt.% relative to the amount of trigonelline, preferably no greater than 0.1 wt.% relative to the amount of trigonelline, more preferably no greater than 0.01 wt.% relative to the amount of trigonelline, most preferably no greater than 0.001 wt.% relative to the amount of trigonelline.

[0074] In an embodiment, at least a portion of the oleuropein is obtained by extraction, e.g., by extraction from a plant such as a plant belonging to the Oleaceae family, preferably one or more of the stems, the leaves, the fruits or the stones of a plant belonging to the Oleaceae family such as Olea europaea (olive tree), a plant of genus Ligustrum, a plant of genus Syringa, a plant of genus Fraximus, a plant of genus Jasminum and a plant of genus Osmanthus. Additionally or alternatively, at least a portion of the oleuropein can be obtained by argan oil, produced from kernels of the argan tree (Argania spinosa) or by chemical synthesis.

[0075] Non-limiting examples of suitable metabolites of oleuropein include oleuropein aglycone, hydroxytyrosol, homovanillyl alcohol, isohomovanillyl alcohol, and mixtures thereof.

[0076] The effective amount of the combination of oleuropein and/or metabolite thereof varies with the particular composition, the age and condition of the recipient, and the particular disorder or disease being treated. Nevertheless, in a general embodiment, 0.001 mg to 1.0 g of the at least one of oleuropein or metabolite thereof can be administered to the individual per day, preferably from 0.01 mg to 0.9 g of the at least one of oleuropein or metabolite thereof per day, more preferably from 0.1 mg to 750 mg of the at least one of oleuropein or metabolite thereof per day, more preferably from 0.5 mg to 500 mg of the at least one of oleuropein or metabolite thereof per day, and most preferably from 1.0 mg to 200 mg of the at least one of oleuropein or metabolite thereof per day

[0077] In some embodiments, the at least one of oleuropein and/or metabolite thereof is the only polyphenol in the composition and/or the only polyphenol administered to the individual.

[0078] Another aspect of the present disclosure is a method of treating or preventing (e.g., reducing incidence and/or severity) a mitochondria-related disease or a condition associated with altered mitochondrial function in an individual human in need thereof or at risk thereof. The method comprises orally administering to a human an effective amount of a combination of trigonelline and at least one of oleuropein and/or metabolite thereof. The mitochondria-related disease or condition can be selected from the group consisting of deleterious effects of aging, stress (e.g., oxidative stress), obesity, overweight, reduced metabolic rate, metabolic syndrome, diabetes mellitus, complications from diabetes, hyperlipidemia, neurodegenerative disease, cognitive disorder, stress-induced or stress-related cognitive dysfunction, mood disorder, anxiety disorder, age-related neuronal death or dysfunction, chronic kidney disease, kidney failure, trauma, infection, cancer, hearing loss, macular degeneration, myopathies and dystrophies, and combinations thereof.

[0079] Yet another aspect of the present disclosure is a method of achieving at least one result selected from the group consisting of (i) improved mitochondrial calcium uptake in one or more cells,

(ii) improved utilization of calcium in one or more cells, (iii) increased mitochondrial energy in one or more cells, (ii) improvement in a physiological state linked to metabolic fatigue in one or more cells,

(iii) treatment or prevention of metabolic fatigue in one or more cells, (iv) treatment or prevention of muscle fatigue, (v) improved mobility, (vi) improved longevity (viii) increasing NAD+ levels in one or more cells and (ix) improved conditions of restrictions of NAD+ bioavailability, the method comprising orally administering to a human an effective amount of a combination of trigonelline and at least one of oleuropein and/or metabolite thereof.

[0080] In an embodiment, the trigonelline and at least one of oleuropein and/or metabolite thereof, are in an amount effective to increase the calcium uptake in mitochondrial cells.

[0081] Aging is a condition that can be linked to one of the following: oxidative stress, reduced level of glutathione, and lower redox ratio NAD⁺/NADH. The compositions disclosed herein can treat or prevent these deleterious effects of aging. For example, trigonelline increases NAD⁺, and the present inventors believe that the increased NAD⁺ may lead to enhancement of glutathione through redox recycling.

[0082] As other examples, depression is linked to low glutathione, and anxiety is linked to oxidative stress. The compositions disclosed herein can treat or prevent these conditions.

[0083] Another aspect of the present disclosure is a unit dosage form of a composition comprising trigonelline and at least one of oleuropein and/or metabolite thereof, and the composition contains the trigonelline and at least one of oleuropein and/or metabolite thereof in an amount effective for treatment or prevention of at least condition selected from the group consisting of deleterious effects of aging, diabetes (type I or type II), complications from diabetes (e.g., diabetic dyslipidemia and/or diabetic microvascular complications such as nephropathy, retinopathy, and/or neuropathy), insulin resistance, metabolic syndrome, dyslipidemia, overweight, obesity, overweight, raised cholesterol levels, raised triglyceride levels, elevated fatty acid levels, fatty liver disease (e.g., non-alcoholic fatty liver disease, including with or without inflammation), cardiovascular disease (e.g., heart failure and/or impaired cardiac contractile function), neurodegenerative disease (e.g., from aging), depression, anxiety, decreased/low motivation, impaired cognitive function, myopathy such as statin-induced myopathy, nonalcoholic steatohepatitis, tinnitus, dizziness, alcohol hangover, hearing impairment, osteoporosis, hypertension, atherosclerosis/coronary artery disease, myocardial damage after stress (e.g., from burns or trauma), traumatic brain injury (including concussions), cystic fibrosis, inflammation, cancer, HIV infection stroke, migraine, and brain ischemia.

[0084] Another aspect of the present disclosure is a method of improving mitochondrial function in a human.

[0085] Yet another aspect of the present disclosure is a method of delaying off-set of metabolic decline, decreasing oxidative stress, maintaining immune function and/or maintaining cognitive function in a healthy older adult. The method comprises administering an effective amount of a composition comprising trigonelline and at least one of oleuropein and/or metabolite thereof to the human.

[0086] Yet another aspect of the present disclosure is a composition to promote neurite outgrowth. In a related embodiment, a method of promoting neurite outgrowth in an individual comprises administering to the individual an effective amount of a composition comprising trigonelline and at least one of oleuropein and/or metabolite thereof to the human.

[0087] Yet another aspect of the present disclosure is a composition comprising trigonelline and at least one of oleuropein and/or metabolite thereof to the human for weight management. “Weight management” for an adult (e.g., at least eighteen years from birth) means that the individual has approximately the same body mass index (BMI) after one week of consumption of the composition, preferably after one month of consumption of the composition, more preferably after one year of consumption of the composition, relative to their BMI when consumption of the composition was initiated. “Weight management” for younger individuals means that the BMI is approximately the same percentile relative to an individual of a corresponding age after one week of consumption of the composition, preferably after one month of consumption of the composition, more preferably after one year of consumption of the composition, relative to their BMI percentile when consumption of the composition was initiated.

[0088] In another aspect, the present disclosure provides a method of improving cognitive function. The method comprises administering an effective amount of acomposition comprising trigonelline and at least one of oleuropein and/or metabolite thereof to the human. The cognitive function can be selected from the group consisting of perception, memory, attention, speech comprehension, speech generation, reading comprehension, creation of imagery, learning, reasoning, and combinations thereof. In an embodiment, the human does not have a cognitive disorder; alternatively, the individual has a cognitive disorder. The human can be elderly and/or can have cognitive decline associated with aging.

[0089] Yet another aspect of the present disclosure is a method of (i) treating or preventing at least one physical state selected from the group consisting of oxidative stress, a condition associated with oxidative stress (e.g., aging and its effects such as skin aging), a reduced level of glutathione, a condition associated with a reduced level of glutathione, or (ii) improving one or more of fetal metabolic programming for prevention of later development of obesity, overweight, pre-diabetes and/or diabetes, maternal and fetal health in gestational diabetes, exercise capacity and physical function, quality of life, longevity, memory, cognition, post-traumatic recovery and survival, or recovery from trauma and surgery. The method comprises administering an effective amount of a composition comprising trigonelline and at least one of oleuropein and/or metabolite thereof to the human at risk thereof or in need thereof. For example, trigonelline increases NAD⁺, and the present inventors believe that the increased NAD⁺ may lead to enhancement of glutathione through redox recycling.

[0090] In biology and psychology, the term "stress" refers to the consequence of the failure of a human to respond appropriately to physiological, emotional, or physical threats, whether actual or imagined. The psychobiological features of stress may present as manifestations of oxidative stress, i.e., an imbalance between the production and manifestation of reactive oxygen species and the ability of a biological system readily to detoxify the reactive intermediates or to repair the resulting damage. Disturbances in the normal redox state of tissues can cause toxic effects through the production of peroxides and free radicals that damage all of the components of the cell, including proteins, lipids, and DNA. Some reactive oxidative species can even act as messengers through a phenomenon called "redox signaling."

[0091] In humans, oxidative stress is involved in many diseases. Examples include atherosclerosis, Parkinson's disease, heart failure, myocardial infarction, Alzheimer's disease, schizophrenia, bipolar disorder, fragile X syndrome, and chronic fatigue syndrome.

[0092] One source of reactive oxygen under normal conditions in humans is the leakage of activated oxygen from mitochondria during oxidative phosphorylation. Other enzymes capable of producing superoxide (O2-) are xanthine oxidase, NADPH oxidases and cytochromes P450. Hydrogen peroxide, another strong oxidizing agent, is produced by a wide variety of enzymes including several oxidases. Reactive oxygen species play important roles in cell signaling, a process termed redox signaling. Thus, to maintain proper cellular homeostasis a balance must be struck between reactive oxygen production and consumption.

[0093] Oxidative stress contributes to tissue injury following irradiation and hyperoxia. It is also suspected to be important in neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), and Huntington's disease.

[0094] Oxidative stress is also thought to be linked to certain cardiovascular diseases, since oxidation of low-density lipoprotein (LDL) in the vascular endothelium is a precursor to plaque formation. Oxidative stress also plays a role in the ischemic cascade due to oxygen reperfusion injury following hypoxia. This cascade includes both strokes and heart attacks. Oxidative stress has also been implicated in chronic fatigue syndrome.

[0095] Moreover, the free radical theory of aging suggests that the biological process of aging results in increased oxidative stress in elderly humans. The ability of a cell to resist the damaging potential of oxidative stress is determined by a vital balance between generation of oxidant free radicals and the defensive array of antioxidants available to the cell. There are multiple antioxidant defense systems and of these, glutathione (GSH) is the most abundant intracellular component of overall antioxidant defenses. GSH, a tripeptide, is synthesized from precursor amino-acids glutamate, cysteine, and glycine in two steps catalyzed by glutamate cysteine ligase (GCL, also known as gamma-glutamylcysteine synthetase, EC 6.3.2.2) and gamma-L-glutamyl-L-cysteine:glycine ligase (also known as glutathione synthetase, EC 6.3.2.3), and GSH synthesis occurs de novo in cells.

[0096] In each of the compositions and methods disclosed herein, the composition is preferably a food product, including food additives, food ingredients, beverage product, functional foods, dietary supplements, medical foods, nutraceuticals, oral nutritional supplements (ONS), food supplements powdered nutritional products to be reconstituted in water or milk before consumption.

[0097] The composition can be administered at least one day per week, preferably at least two days per week, more preferably at least three or four days per week (e.g., every other day), most preferably at least five days per week, six days per week, or seven days per week. The time period of administration can be at least one week, preferably at least one month, more preferably at least two months, most preferably at least three months, for example at least four months. In some embodiments, dosing is at least daily; for example, a subject may receive one or more doses daily, in an embodiment a plurality of doses per day. In some embodiments, the administration continues for the remaining life of the individual. In other embodiments, the administration occurs until no detectable symptoms of the medical condition remain. In specific embodiments, the administration occurs until a detectable improvement of at least one symptom occurs and, in further cases, continues to remain ameliorated.

[0098] The compositions disclosed herein may be administered to the subject enterally, e.g., orally, or parenterally. Non-limiting examples of parenteral administration include intravenously, intramuscularly, intraperitoneally, subcutaneously, intraarticularly, intrasynovially, intraocularly, intrathecally, topically, and inhalation. As such, non-limiting examples of the form of the composition include natural foods, processed foods, natural juices, concentrates and extracts, injectable solutions, microcapsules, nano-capsules, liposomes, plasters, inhalation forms, nose sprays, nosedrops, eyedrops, sublingual tablets, and sustained-release preparations.

[0099] The compositions disclosed herein can use any of a variety of formulations for therapeutic administration. More particularly, pharmaceutical compositions can comprise appropriate pharmaceutically acceptable carriers or diluents and may be formulated into preparations in solid, semi-solid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, gels, microspheres, and aerosols. As such, administration of the composition can be achieved in various ways, including oral, buccal, rectal, parenteral, intraperitoneal, intradermal, transdermal, and intratracheal administration. The active agent may be systemic after administration or may be localized by the use of regional administration, intramural administration, or use of an implant that acts to retain the active dose at the site of implantation.

[00100] In pharmaceutical dosage forms, the compounds may be administered as their pharmaceutically acceptable salts. They may also be used in appropriate association with other pharmaceutically active compounds. The following methods and excipients are merely exemplary and are in no way limiting.

[00101] For oral preparations, the compounds can be used alone or in combination with appropriate additives to make tablets, powders, granules or capsules, for example, with conventional additives, such as lactose, mannitol, corn starch or potato starch; with binders, such as crystalline cellulose, cellulose functional derivatives, acacia, corn starch or gelatins; with disintegrators, such as corn starch, potato starch or sodium carboxymethylcellulose; with lubricants, such as talc or magnesium stearate; and if desired, with diluents, buffering agents, moistening agents, preservatives and flavoring agents.

[00102] The compounds can be formulated into preparations for injections by dissolving, suspending or emulsifying them in an aqueous or non-aqueous solvent, such as vegetable or other similar oils, synthetic aliphatic acid glycerides, esters of higher aliphatic acids or propylene glycol; and if desired, with conventional, additives such as solubilizers, isotonic agents, suspending agents, emulsifying agents, stabilizers and preservatives. [00103] The compounds can be utilized in an aerosol formulation to be administered by inhalation. For example, the compounds can be formulated into pressurized acceptable propellants such as dichlorodifluoromethane, propane, nitrogen and the like.

[00104] Furthermore, the compounds can be made into suppositories by mixing with a variety of bases such as emulsifying bases or water-soluble bases. The compounds can be administered rectally by a suppository. The suppository can include a vehicle such as cocoa butter, carbowaxes and polyethylene glycols, which melt at body temperature, yet are solidified at room temperature.

[00105] Unit dosage forms for oral or rectal administration such as syrups, elixirs, and suspensions may be provided wherein each dosage unit, for example, teaspoonful, tablespoonful, tablet or suppository, contains a predetermined amount of the composition. Similarly, unit dosage forms for injection or intravenous administration may comprise the compounds in a composition as a solution in sterile water, normal saline or another pharmaceutically acceptable carrier, wherein each dosage unit, for example, mL or L, contains a predetermined amount of the composition containing one or more of the compounds.

[00106] EXAMPLE

[00107] Comparative effect of the combination of Oleuropein aglycone plus Trigonelline vs single compounds on mitochondrial activation in muscle cells

[00108] Materials and Methods

[00109] To test the effect of Oleuropein aglycone (Oea), Trigonelline and their combination on mitochondrial activation in living cells, the inventors measured mitochondrial calcium elevation in myotubes differentiated from C2C12 cells.

[00110] C2C12 cells were purchased from ATCC. C2C12 cells were seeded in 384-well plates at a density of 4500 cells per well in DMEM medium, high glucose (Gibco) + 10% fetal calf serum. Myotubes were differentiated from C2C12 cells by growing the cells in DMEM containing 2% horse serum, for 6 days.

[00111] Mitochondrial calcium measurements were carried out using myotubes infected with the adenovirus (from Sirion biotech) expressing the luminescent mitochondrially-targeted calcium sensor mitochondrial mutated aequorin (Montero et al., 2000). For aequorin reconstitution, 48 hours after infection, myotubes were incubated for 2h at room temperature (22 ±°C) in standard Aequorin buffer (145 mM NaCl, 5 mM KC1, 1 mM MgC12, 1 mM CaC12, 10 mM glucose and 10 mM Hepes, pH 7.4) with 1 pM wild type coelenterazine. For treatment, Trigonelline Iodine (0 pM, buffer, or 500 pM„ as indicated in the figures) and Oleuropein aglycone (3 pM or 10 pM, as indicated in the figures) were incubated for 2h in Aequorin buffer. Myotubes were stimulated with 5 mM of caffeine and the total calcium transiting during stimulation was calculated as the area under the curve, during caffeine stimulation. Luminescence was measured at the FLIPR cell imaging reader (Molecular devices). Calibration of the luminescence data into Calcium concentration was carried out using an algorithm as described previously (Bonora et al., 2013). Custom module analysis based on Excel (Microsoft) and GraphPad Prism 7.02 (GraphPad) software was used for quantification.

[00112] Results

[00113] As shown in FIG. 1 and FIG.2, the combination of Trigonelline and Oleuropein promotes the beneficial effects of the 2 compounds by boosting the calcium uptake into the mitochondria.

[00114] REFERENCES

[00115] Glancy, B. and R. S. Balaban (2012). "Role of mitochondrial Ca2+ in the regulation of cellular energetics." Biochemistry 51(14): 2959-2973.

[00116] Montero, M., Alonso, M.T., Carnicero, E., Cuchillo-Ibanez, I., Albillos, A., Garcia, A.G, Garcia-Sancho, J., & Alvarez, J. (2000), Chromaffin-cell stimulation triggers fast millimolar mitochondrial Ca²⁺ transients that modulate secretion, Nature Cell Biology, 2, 57-61.

[00117] Bonora, M., Giorgi, C., Bononi, A., Marchi, S., Patergnani, S., Rimessi, A., Rizzuto, R., and Pinton, P. (2013). Subcellular calcium measurements in mammalian cells using jellyfish photoprotein aequorin-based probes. Nature Protocols 8, 2105-2118.

[00118] It should be understood that various changes and modifications to the presently preferred embodiments described herein will be apparent to those skilled in the art. Such changes and modifications can be made without departing from the spirit and scope of the present subject matter and without diminishing its intended advantages. It is therefore intended that such changes and modifications be covered by the appended claims.

Claims

1. A method of treating or preventing a mitochondria-related disease or a condition associated with altered mitochondrial function in an individual in need thereof or at risk thereof, the method comprising orally administering to the individual an effective amount of a combination of trigonelline and at least one of oleuropein and/or metabolite thereof.

2. The method of Claim 1, wherein the mitochondria-related disease or condition is selected from the group consisting of deleterious effects of aging, stress, obesity, overweight, reduced metabolic rate, metabolic syndrome, diabetes mellitus, complications from diabetes, hyperlipidemia, neurodegenerative disease, cognitive disorder, stress-induced or stress-related cognitive dysfunction, mood disorder, anxiety disorder, age-related neuronal death or dysfunction, chronic kidney disease, kidney failure, trauma, infection, cancer, hearing loss, macular degeneration, myopathies and dystrophies, and combinations thereof.

3. A method of achieving at least one result selected from the group consisting of (i) improved mitochondrial calcium uptake in one or more cells, (ii) improved utilization of calcium in one or more cells, (iii) increased mitochondrial energy in one or more cells, (ii) improvement in a physiological state linked to metabolic fatigue in one or more cells, (iii) treatment or prevention of metabolic fatigue in one or more cells, (iv) treatment or prevention of muscle fatigue, (v) improved mobility, (vi) improved longevity (viii) increasing NAD+ levels in one or more cells and (ix) improved conditions of restrictions of NAD+ bioavailability, the method comprising orally administering to a human an effective amount of a combination of trigonelline and at least one of oleuropein and/or metabolite thereof.

4. The method of any of Claims 1 to 3, wherein the trigonelline and at least one of oleuropein and/or metabolite thereof, is in an amount effective to increase the calcium uptake in mitochondrial cells.

5. The method of any of Claim 1 to 4, wherein the composition is selected from the group consisting of a food product, a beverage product, a food supplement, an oral nutritional supplement (ONS), a medical food, powdered nutritional products to be reconstituted in water or milk before consumption and combinations thereof.

6. The method of any one of preceding claims, wherein the one or more cells are part of at least one body part selected from the group consisting of a liver, a kidney, a brain, and a skeletal muscle.

7. The method of any one of preceding claims, wherein the composition is formulated for enteral administration.

8. The method according to any one of preceding claims, wherein at least a portion of trigonelline is isolated.

9. The method according to any one of preceding claims, wherein at least a portion of trigonelline is provided by a plant or algae extract in the composition.

10. The method according to any one of preceding claims, wherein at least a portion of trigonelline is provided by a trigonelline-enriched plant or algae extract in the composition.

11. The method according to any one of preceding claims, wherein the trigonelline is selected from an extract of coffee, fenugreek or algae.

12. The method according to any one of preceding claims, wherein trigonelline is selected from an extract of fenugreek which contains at least about 25% - 50% trigonelline.

13. The composition according to any one of preceding claims, wherein trigonelline is chemically synthesized and which contains at least about 90% trigonelline.

14. The method of any of preceding claims, wherein the metabolite of oleuropein is selected from the group consisting of oleuropein aglycone, hydroxytyrosol, homovanillyl alcohol, isohomovanillyl alcohol, glucuronidated forms thereof, sulfated forms thereof, derivatives thereof and mixtures thereof.

15. The method of any of preceding claims, wherein the combination of trigonelline and at least one of oleuropein and/or metabolite thereof is administered daily for at least one week.

16. The method of any of preceding claims, wherein the trigonelline and the at least one of oleuropein and/or metabolite thereof are administered together in the same composition.

17. The method of any of preceding claims, wherein the trigonelline is administered separately in a different composition from the at least one of oleuropein and/or metabolite thereof.