Compositions and methods for treatment of "plaques and tangles" in humans and animals

Abstract

To provide compositions and methods for treating brain "plaques" and / or "tangles" in a subject.SOLUTION: This method comprises administering a therapeutically effective amount of a composition comprising a black currant extract, an Uncaria tomentosa extract, and an oolong tea extract.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] Related Applications This application claims priority benefit of U.S. Provisional Application No. 62 / 778,875, filed December 12, 2018, the entire contents of which are incorporated herein by reference. Technical Field

[0002] The present invention relates to a blended composition of blackcurrant, Uncaria tomentosa, and oolong tea plant extracts, and methods of using such compositions to treat "plaques" and "tangles" that accumulate in the aging brain in amyloidosis and tauopathy in humans and animals (such as elderly dogs and cats). Furthermore, the present invention relates to the development of a dietary supplement blend composition comprising a combination of blackcurrant extract, Uncaria tomentosa plant extract, and oolong tea extract for preventing and treating traumatic brain injury (TBI), concussion (TBI), single and repeated blows to the head (observed in most athletes and military / soldiers), and chronic traumatic encephalopathy (CTE). Finally, the present invention relates to the surprising discovery that the blended composition is more soluble than a blended composition containing Uncaria tomentosa and oolong tea but not blackcurrant extract. [Background technology]

[0003] Beta-amyloid plaque accumulation in the brain in amyloidosis and various disorders Alzheimer's disease is characterized by the accumulation of a fibrillar form of a 39-43 amino acid peptide called beta amyloid protein, or Aβ, which exists as extracellular amyloid plaques and as amyloid within the walls of cerebral blood vessels. Fibrillar Aβ amyloid deposition in Alzheimer's disease is believed to be detrimental to patients and ultimately leads to toxicity and neuronal cell death, the hallmark features of Alzheimer's disease. Accumulating evidence implicates amyloid, more specifically, the formation, deposition, accumulation, and / or persistence of Aβ fibrils, as a major causative factor in the pathogenesis of Alzheimer's disease. Furthermore, in addition to Alzheimer's disease, several other amyloid diseases, including Down syndrome, disorders associated with congophilic vasculopathy such as hereditary cerebral hemorrhage of the Dutch type, inclusion body myositis, dementia pugilistica, cerebral beta-amyloid angiopathy, dementia associated with progressive supranuclear palsy, and dementia associated with corticobasal degeneration and mild cognitive impairment, also involve the formation, deposition, accumulation, and persistence of Aβ fibrils.

[0004] Amyloid diseases (amyloidoses) are classified according to the type of amyloid protein and the underlying disease. Amyloid diseases share several common characteristics, including each amyloid consisting of a unique type of amyloid protein. Amyloid diseases include, but are not limited to, Alzheimer's disease, Down's syndrome, canine dysfunction syndrome (CDS), amyloid associated with canine cognitive dysfunction (CCD) (as seen in older animals such as dogs and cats), hereditary cerebral hemorrhage with Dutch-type amyloidosis, dementia pugilistica, inclusion body myositis (Askanas et al., Ann. Neurol. 43:521-560, 1993), and mild cognitive impairment (the specific amyloid is called beta-amyloid protein, or Aβ), amyloid associated with chronic inflammation, various forms of malignancy and familial Mediterranean fever (the specific amyloid is called AA amyloid or inflammation-associated amyloidosis), amyloid associated with multiple myeloma and other B-cell cachexia (the specific amyloid is called AL amyloid), type 2 amyloidosis, and the like. These include amyloids associated with diabetes (the specific amyloid is called amylin or islet amyloid polypeptide, or IAPP), amyloids associated with prion diseases such as Creutzfeldt-Jakob disease, Gerstmann-Straussler syndrome, kuru, and animal scrapie (the specific amyloid is called PrP amyloid), amyloids associated with long-term hemodialysis and carpal tunnel syndrome (the specific amyloid is called α2-microglobulin amyloid), amyloids associated with senile cardiac amyloidosis and familial amyloidotic polyneuropathy (the specific amyloid is called transthyretin or prealbumin), and amyloids associated with endocrine tumors such as medullary thyroid carcinoma (the specific amyloid is called a variant of procalcitonin).Furthermore, α-synuclein protein, which forms amyloid-like fibrils and is Congo red and thioflavin S positive (specific stains used to detect amyloid fibril deposits), has been implicated in Parkinson's disease, Lewy body disease (Lewy in Handbuch der Neurologie, M. Lewandowski, ed., Springer, Berlin, pp. 920-933, 1912; Pollanen et al., J. Neuropath. Exp. Neurol. 52:183-191, 1993; Spillantini et al., Proc. Natl. Acad. Sci. USA 95:6469-6473, 1998; Arai et al., Neurosc. Lett. 259:83-86, 1999), multiple system atrophy (Wakabayashi et al., Acta Neuropath. 96:445-452, 1998), dementia with Lewy bodies, and Alzheimer's disease with Lewy bodies are found as part of Lewy bodies in the brains of patients with Alzheimer's disease. For the purposes of this disclosure, Parkinson's disease is now considered a disease that also exhibits features of amyloid disease due to the fact that fibrils occur in the brains of patients with this disease (which are Congo red and thioflavin S positive and contain a predominant beta-pleated sheet secondary structure). Amyloid as a therapeutic target for Alzheimer's disease

[0005] Alzheimer's disease is characterized by the deposition and accumulation of a 39-43 amino acid peptide called beta-amyloid protein, Aβ, or β / A4 (Glenner and Wong, Biochem. Biophys. Res. Comm. 120:885-890, 1984; Masters et al., Proc. Natl. Acad. Sci. USA 82:4245-4249, 1985; Husby et al., Bull. WHO 71:105-108, 1993). Aβ is derived by protease cleavage from a larger precursor protein called β-amyloid precursor protein (APP), which has several alternatively spliced ​​variants. The most abundant forms of APP contain proteins of 696, 751, and 770 amino acids (Tanzi et al., Nature 31:528-530, 1980).

[0006] Small Aβ peptides are the major component of amyloid deposits, or "plaques," in the brains of Alzheimer's disease patients. Furthermore, Alzheimer's disease is characterized by the presence of numerous neurofibrillar "tangles," consisting of paired helical filaments, that abnormally accumulate in neuronal cytoplasm (Grundke-Iqbal et al., Proc. Natl. Acad. Sci. USA 83:4913-4917, 1986; Kosik et al., Proc. Natl. Acad. Sci. USA 83:4044-4048, 1986; Lee et al., Science 251:675-678, 1991). Therefore, the pathological hallmark of Alzheimer's disease is the presence of both "plaques" and "tangles," with amyloid deposits in the central core of the plaques. Another major type of lesion found in the brain of Alzheimer's disease is the accumulation of amyloid in the walls of blood vessels, both within the brain parenchyma and in the walls of the middle meningeal blood vessels outside the brain. Amyloid deposits localized in the walls of blood vessels are called cerebrovascular amyloid or congophilic vasculopathy (Mandybur, J. Neuropath. Exp. Neurol. 45:79-90, 1986; Pardridge et al., J. Neurochem. 49:1394-1401, 1987).

[0007] For many years, scientific debate has been ongoing regarding the importance of amyloid in Alzheimer's disease and whether the characteristic plaques and tangles are the cause or merely a consequence of the disease. Over the past few years, research has demonstrated that amyloid is indeed a causative factor in Alzheimer's disease and should not be viewed as merely an innocent bystander. Alzheimer's Aβ protein in cell culture has been shown to cause neuronal degeneration within a short period of time (Pike et al., Br. Res. 563:311-314, 1991; J. Neurochem. 64:253-265, 1995). Research has shown that the neurotoxic effect is due to the fibrillar structure (consisting of a predominantly beta-pleated sheet secondary structure) characteristic of all amyloids. Aβ is also neurotoxic in hippocampal slice cultures (Harrigan et al., Neurobiol. Aging 16:779-789, 1995) and has been found to induce neuronal cell death in transgenic mice (Games et al., Nature 373:523-527, 1995; Hsiao et al., Science 272:99-102, 1996). Injection of Alzheimer's disease Aβ into rat brains also causes memory impairment and neuronal dysfunction (Flood et al., Proc. Natl. Acad. Sci. USA 88:3363-3366, 1991; Flood et al., Br. Res. 663:271-276, 1994).

[0008] Perhaps the most compelling evidence that Aβ amyloid is directly involved in the pathogenesis of Alzheimer's disease comes from genetic studies. It has been discovered that the production of Aβ can result from mutations in the gene encoding its precursor, the beta-amyloid precursor protein (Van Broeckhoven et al., Science 248:1120-1122, 1990; Murrell et al., Science 254:97-99, 1991; Haass et al., Nature Med. 1:1291-1296, 1995). The identification of mutations in the beta-amyloid precursor protein gene that cause early-onset familial Alzheimer's disease is the strongest argument that amyloid is central to the pathogenic process underlying this disease. Several reported disease-causing mutations have been discovered, demonstrating the importance of Aβ in causing familial Alzheimer's disease (reviewed in Hardy, Nature Genet. 1:223-234, 1992). All of these studies suggest that providing therapies, drugs, or supplements to reduce, eliminate, and / or prevent the formation, deposition, accumulation, and / or persistence of fibrillar Aβ in the human and animal brains may well serve as an effective treatment. Accumulation of "plaques and tangles" in the aging human and animal brains

[0009] The human brain is the most complex organ in the universe. It weighs only 3 pounds, or about 2% of body weight. Yet, it consumes 20-30% of the calories we burn, 20% of the oxygen we breathe, and 25% of the blood flow in the body, and is composed of 85% water (Daniel G. Amen, MD, 12 prescriptions for creating a brain healthy life. Source: www.amenclinics.com / cybcyb / 12-prescriptions-for-creating-a-brain-healthy-life / ). The brain contains approximately 100 billion nerve cells (i.e., neurons) and up to 4 trillion connections called synapses (ibid.). As the human brain ages, it loses approximately 85,000 cortical neurons per day, or about one every second. (Deepak Chopra, MD, and Rudolph Tanzi, Ph.D. Super Brain. Unleashing the Explosive Power of Your Mind to Maximize Health, Happiness, and Spiritual Well Being. See: / / / www.chopra.com / super-brain-by-deepak-chopra-rudolph-tanzi.) As the brain ages, there is a slow but deliberate accumulation of two neurotoxic proteins, beginning in the 20s. The first is the accumulation in the brain of an insoluble (aggregated) specific neurotoxic protein known as "beta-amyloid protein," or Aβ. Beta-amyloid protein deposits in the form of "plaques" (which look like "meatballs" in the brain under a microscope) have been shown to aid in killing healthy neurons, leading to the decline of hippocampus-dependent memory and cognition. Dr. Alan Snow and co-inventors have developed patented methods for generating "in vitro plaque" (similar to that found in the human brain) and have used these methods to screen and identify natural "plaque-reducing" dietary supplement ingredients (U.S. Patent No. 7,148,001, which is incorporated herein by reference in its entirety).

[0010] A second neurotoxic protein that accumulates in the aging brain is known as "tau protein," which forms twisted, paired helical filaments known as "tangles." Neurofibrillary tangles accumulate inside neurons, causing them to die and appearing under a microscope like "dried spaghetti strands." Dr. Snow's lab developed proprietary methods for forming "tangles" in test tubes and then used these assays to identify dietary supplement ingredients that "inhibit tangles" (see example below). Thus, in the aging brain, both "plaques and tangles" accumulate, causing neurons to die and disrupting connections between nerve cells (called synapses), leading to a gradual decline in memory and cognition. Compounds or agents that can disaggregate and reduce the accumulation of "plaques and tangles" have been shown to improve memory and reduce memory decline (Karinoski et al., Suppression of amyloid deposition leads to long-term reductions in Alzheimer's pathologies in Tg2576 mice. J. Neurosc. 29:4964-4971, 2009; Vellas et al., Long-term follow-up of patients immunized with AN1792: Reduced functional decline in antibody responders. Current Alz. Res. 6:144-151, 2009; Morgan et al., Aβ peptide vaccination prevents memory loss in an animal model of Alzheimer's disease. Nature 408:982-985, 2000; Chen et al., A learning deficit related to age and β-amyloid plaques in a mouse model of Alzheimer's disease.Nature 408:975 - 979, 2000; Janus et al., Aβ peptide immunization reduces behavioral impairment and plaques in a model of Alzheimer’s disease. Nature 408:979 - 985, 2000; Schenk et al., Immunization with amyloid-β attenuates Alzheimer-disease like pathology in PDAPP mouse. Nature 400:173 - 177, 1999; Yanamandra et al., Anti-tau antibodies that block tau aggregate seeding in vitro markedly decreases pathology and improves cognition in vivo. Neuron 80:402 - 414, 2013; Dumont et al., Bezafibrate administration improves behavioral deficits and tau pathology in P3015 mice. Human Molecular Genetics 21:5091 - 5105, 2012; Oddo et al., Reduction of soluble Abeta and tau, but not soluble Abeta alone, ameliorates cognitive decline in transgenic mice with plaques and tangles. J. Biol. Chem. 281:39413 - 39423, 2006; Santacruz et al., Tau suppression in a neurodegenerative mouse model improves memory function. Science 309:476 - 481, 2005).

[0011] The only difference between aging brains that may lead to age-associated memory impairment (AAMI), mild cognitive impairment (MCI), and potentially Alzheimer's disease, and non-aging brains is the number of "plaques and tangles" in the brain. Brains with Alzheimer's disease have tens to hundreds of thousands of "plaques and tangles" per square millimeter, which significantly increases neuronal death and leads to loss of synapses (connections between neurons), resulting in memory loss and cognitive decline.

[0012] Thus, beta-amyloid and tau are two key proteins in the aging brain that accumulate as insoluble "plaques and tangles" and have been shown to be directly associated with memory loss and cognitive decline. Currently, there are no approved medications to reduce and remove both beta-amyloid protein "plaques" and tau protein-containing "tangles" in the brain. Plaque buildup in the brains of aging dogs and cats

[0013] Dogs and cats also accumulate "plaques" (and, to a lesser extent, "tangles") in their brains as they age, which are thought to contribute to memory decline and cognitive impairment. The same beta-amyloid proteins (i.e., "plaques") and tau proteins (i.e., "tangles") that accumulate in the human brain have been shown to accumulate in elderly dogs (Papoiannou et al., Immunohistochemical investigation of the brain of aged dogs. I. Detection of neurofibrillary tangles and of 4-hydroxynonenal protein, an oxidative damage product, in senile plaques. Amyloid 8:11-21, 2001; Uchida et al., Amyloid angiopathy with cerebral hemorrhage and senile plaques in aged dogs. Nihon Juigaku Zasshi 52:605-11, 1990) and cats (Gunn-Moore et al., Cognitive dysfunction and the neurobiology of aging in cats. J Small Anim. Pract. 48:546-53, 2007; Nakamura et al., Senile plaques in very aged dogs. It also accumulates in cats. Acta Neuropath. 91:437-9, 1996).

[0014] Canine cognitive dysfunction (CCD) (also known as cognitive dysfunction syndrome or CDS) is a widespread disease in dogs and cats that manifests as dementia or Alzheimer's disease in humans. CCD causes pathological changes in the brain, impairing mental function in dogs and cats, resulting in loss of memory, motor skills, and behaviors learned through training earlier in life. In dog and cat brains, the cause is again beta-amyloid protein, or Aβ, which forms "plaques" in the brain. As dogs age, more and more "plaques" accumulate, causing nerve cell death. While the initial symptoms of the disorder are mild, they gradually worsen over time, also known as "cognitive decline." Amyloid "plaques" develop in older dogs at approximately 5-7 years of age and in cats at approximately 10 years of age (corresponding to an average lifespan of 15-20 years). In fact, clinical signs of cognitive dysfunction syndrome are present in 50% of dogs over the age of 11, and by age 15, 68% of dogs will exhibit at least one sign. Dogs often become disoriented in familiar areas of the home, spend long periods of time in one area of ​​the home, do not respond to calls or commands, and experience abnormal sleep patterns.

[0015] Plaques containing beta-amyloid protein have also been identified in the brains of other higher mammals, including monkeys, bears, camels, and horses (Nakamura et al., Histopathological studies of senile plaques and cerebral amyloidosis in cynomolgus monkeys. J Med Primatol. 27:244-52, 1998; Capucchio et al., studies. J Comp Pathol 142:61-73, 2010; Nakamura et al., Senile plaques in an aged two-humped (Bactrian) camel (Camelus bactrianus), Acta Neuropathol 90:415-8, 1995; Uchida et al., Senile plaques and other senile changes in the brain of an aged American black bear, Vet. Pathol. 32:412-4, 1995). Tauopathies and "tangles"

[0016] Tau was discovered in the mid-1970s as a microtubule-associated protein (Weingarten, 1975). In addition to being a stabilizer of microtubules in neurons and other cells, it has since been found to play an important role in cell differentiation, polarization, and axonal transport. Normal tau is a soluble protein bound to microtubules, but in a range of neurodegenerative diseases now known as tauopathies, tau accumulates as a pathogenic insoluble fibrillar protein. These tau inclusions appear to modulate the severity of dementia and the clinical features of these neurodegenerative diseases. Tauopathies include Alzheimer's disease, frontotemporal lobar degeneration with tau inclusions (FTLD-tau) such as Pick's disease, progressive supranuclear palsy, and diseases such as corticobasal degeneration, argyrophilic grain dementia, several prion diseases, amyotrophic lateral sclerosis / parkinsonism-dementia complex, chronic traumatic encephalopathy, and some genetic forms of Parkinson's disease (V.M. et al., Ann. Rev. Neurosci. 24:1121-1159, 2001; B. Omalu et al., Neurosurgery 69(1):173-83, 2011; A. Rajput et al., Neurology 67:1506-1508, 2006; G. Santpere and I. Ferrer, Acta Neuropathol. 117:227-246, 2009).

[0017] One of the most striking effects of the buildup of fibrillar tau in the brain is a gradual deterioration of short-term memory, i.e., the ability to quickly recall recently stored memories (P. Giannakopoulos et al., Neurology 60:1495-1500, 2003). Because there is no treatment for these disorders, it is important to find novel methods that can target this pathogenic protein and improve memory impairment. "Tangles" accumulate in the brain in traumatic brain injury (TBI), concussion, head injury, and chronic traumatic encephalopathy (CTE)

[0018] Brain "tangles" of tau protein also gradually accumulate in the brain after blows to the head, such as concussions, head trauma, post-traumatic stress disorder (PTSD), and blast traumatic brain injury (seen in soldiers and military personnel with traumatic head injuries caused by a single blast). The movie "Concussion" and / or the NFL Players Association all describe dementia-like behaviors seen in athletes after repeated concussions and head blows (known as traumatic brain injury, or TBI). Loss of consciousness is a clinical feature of concussion but is not required to make a diagnosis. Other symptoms include confusion, disorientation, unsteadiness, dizziness, headache, and visual disturbances.

[0019] The long-term effects of traumatic brain injury are called chronic traumatic encephalopathy (CTE), a form of tauopathy (i.e., the formation of "tangles" containing tau proteins in the brain). CTE is a progressive degenerative disease seen in people who have suffered repeated brain trauma, including subconcussive blows to the head, that do not cause immediate symptoms. This disease was first discovered in people with a history of boxing and was therefore formerly known as dementia pugilistica (DP), or "punch-drunk." CTE is now found in the brains of professional athletes, including NFL football players, ice hockey, rugby, skiing, skateboarding, stunt performers, bull riding, rodeo, and all other contact sports that experience repeated brain trauma. Patients with CTE exhibit many signs of dementia, such as memory loss, aggression, confusion, and depression, which can appear years or even decades after the trauma. In September 2015, researchers from the Department of Veterans Affairs and Boston University announced that they had identified CTE in 96% of NFL football players they surveyed and in 79% of all football players (Jason Breslow, New: 87 deceased NFL players test positive for brain disease. Frontline, January 9, 2016).

[0020] Under a microscope, neuropathology is evident, primarily consisting of accumulations of tau protein "tangles," similar to those found in the brains of Alzheimer's disease patients. While some beta-amyloid protein deposits (i.e., "plaques") are also present, these are typically rare and less distinctive than the accumulation of brain "tangles." These findings suggest that a blow to the head can result in the near-instant accumulation of brain "tangles," which can lead to dementia-like symptoms, such as memory loss and cognitive decline. Identifying nutritional supplements that can help reduce and / or remove brain "tangles" would likely be specialized supplements taken daily by athletes of all kinds, NFL players, the military, and their soldiers. Summary of the Invention [Problem to be solved by the invention]

[0021] In one aspect, provided is a composition comprising a therapeutically effective amount of a blackcurrant extract, an Uncaria tomentosa extract, and an oolong tea extract.

[0022] In some embodiments, the blackcurrant extract comprises about 10% to about 35% w / w of proanthocyanidins, anthocyanidins, anthocyanins, or a combination thereof. In some embodiments, the blackcurrant extract comprises about 25% w / w of proanthocyanidins, anthocyanidins, anthocyanins, or a combination thereof. In some embodiments, the composition comprises about 10 mg to about 100 mg of blackcurrant extract. In some embodiments, the composition comprises about 1% to about 10% w / w of blackcurrant extract. In some embodiments, the composition comprises about 100 mg to about 500 mg of Uncaria tomentosa extract. In some embodiments, the composition comprises about 10% to about 40% w / w of Uncaria tomentosa extract. In some embodiments, the composition comprises about 100 mg to about 500 mg of oolong tea extract. In some embodiments, the composition comprises about 10% to about 40% w / w of oolong tea extract. In some embodiments, the composition is formulated as a pill, tablet, caplet, soft or hard gelatin capsule, lozenge, sachet, cachet, veggie cap, drop, elixir, suspension, emulsion, solution, beverage preparation, cold or hot tea beverage, syrup, tea bag, aerosol, suppository, sterile injectable solution, or sterile packaged powder. In some embodiments, the composition is formulated as a capsule. In some embodiments, the capsule is about 200 mg to about 1000 mg.

[0023] In another aspect, a composition is provided comprising a therapeutically effective amount of blackcurrant extract.

[0024] In some embodiments, the blackcurrant extract comprises about 10% to about 35% w / w of proanthocyanidins, anthocyanidins, anthocyanins, or a combination thereof. In some embodiments, the blackcurrant extract comprises about 25% w / w of proanthocyanidins, anthocyanidins, anthocyanins, or a combination thereof. In some embodiments, the composition comprises about 10 mg to about 100 mg of blackcurrant extract. In some embodiments, the composition comprises about 1% to about 10% w / w of blackcurrant extract. In some embodiments, the composition further comprises a therapeutically effective amount of Uncaria tomentosa extract. In some embodiments, the composition comprises about 100 mg to about 500 mg of Uncaria tomentosa extract. In some embodiments, the composition comprises about 10% to about 40% w / w of Uncaria tomentosa extract. In some embodiments, the composition further comprises a therapeutically effective amount of oolong tea extract. In some embodiments, the composition comprises about 100 mg to about 500 mg of oolong tea extract. In some embodiments, the composition comprises about 10% to about 40% w / w oolong tea extract. In some embodiments, the composition is formulated as a pill, tablet, caplet, soft or hard gelatin capsule, lozenge, sachet, cachet, veggie cap, drop, elixir, suspension, emulsion, solution, beverage preparation, cold or hot tea beverage, syrup, tea bag, aerosol, suppository, sterile injectable solution, or sterile packaged powder. In some embodiments, the composition is formulated as a capsule. In some embodiments, the capsule is about 200 mg to about 1000 mg. In some embodiments, the composition has higher solubility than a composition comprising Uncaria tomentosa extract, oolong tea extract, and no blackcurrant extract.

[0025] In another aspect, there is also provided a method for the treatment of amyloidosis in a mammal in need thereof, comprising administering to a mammal in need thereof any one of the compositions described herein.

[0026] In some embodiments, the amyloidosis is Alzheimer's disease, Down's syndrome, dementia pugilistica, cognitive impairment syndrome, canine cognitive impairment, multiple system atrophy, inclusion body myositis, hereditary cerebral hemorrhage with Dutch amyloidosis, Niemann-Pick disease type C, cerebral amyloid angiopathy, dementia associated with corticobasal degeneration, amyloidosis in type 2 diabetes, amyloidosis in chronic inflammation, amyloidosis in malignancy and familial Mediterranean fever, amyloidosis in multiple myeloma and B-cell dyscrasia, amyloidosis in prion disease, Creutzfeldt-Jakob disease, Gerstmann-Straussler syndrome, amyloidosis associated with kuru, scrapie, carpal tunnel syndrome, senile cardiac amyloidosis, familial amyloidotic polyneuropathy, or amyloidosis associated with endocrine tumors.

[0027] In another embodiment, a method of treating amyloid fibril formation, deposition, accumulation, or persistence is provided, comprising treating the fibrils with any one of the compositions described herein.

[0028] In another aspect, a method of treating the formation, deposition, accumulation, or persistence of beta-amyloid-containing plaques is provided, comprising treating the plaques with any one of the compositions described herein.

[0029] In another aspect, there is provided a method for treating a tauopathy in a human or mammal in need thereof, the method comprising administering any one of the compositions described herein to the human or mammal in need thereof.

[0030] In some embodiments, the tauopathy is Alzheimer's disease, frontotemporal lobar degeneration with inclusions (FLTD-tau), Pick's disease, progressive supranuclear palsy, corticobasal degeneration, argyrophilic grain dementia, prion disease, amyotrophic lateral sclerosis-parkinsonism-dementia complex of Guam (also known as Richkovaig's disease), Parkinson's disease, tangle-predominant dementia, ganglion, ganglioglioma, gangliocytoma, meningiomatosis, subacute sclerosing panencephalitis, lead encephalopathy, tuberous sclerosis, Hallervorden-Spatz disease, lipofuscinosis, traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE), dementia pugilistica, concussion, single or repeated blows to the head, or post-traumatic stress disorder.

[0031] In another aspect, a method of treating the formation, deposition, accumulation, or persistence of tangles comprising tau protein, comprising treating the tangles with any of the compositions described herein.

[0032] In another embodiment, a method of treating the formation, deposition, accumulation, or persistence of tau protein-containing plaques comprises treating the plaques with any one of the compositions described herein.

[0033] In one aspect, a method for improving cognitive performance and / or slowing cognitive decline in a subject in need thereof, the method comprising administering to the subject any one of the compositions described herein to reduce the formation, deposition, accumulation or persistence of beta-amyloid protein plaques or fibrils in the subject.

[0034] In some embodiments, the subject is a patient suffering from an amyloid disease.

[0035] Also provided in one aspect is a method for improving learning, memory, cognition, concentration and / or attention in a subject in need thereof, comprising administering to the subject any one of the compositions described herein.

[0036] In some embodiments, the subject is a patient with age-associated memory impairment (AAMI), mild cognitive impairment (MCI), or Alzheimer's disease.

[0037] In another aspect, provided is a method for inhibiting, preventing, reducing, or treating Tangles in the brain of a subject in need thereof, the method comprising administering to the subject any one of the compositions described herein.

[0038] In some embodiments, the subject is a patient who has experienced single or multiple concussions, traumatic brain injury (TBI), a blow to the head, chronic traumatic encephalopathy (CTE), post-traumatic stress disorder, brain aging, mild cognitive impairment, or Alzheimer's disease.

[0039] In another aspect, a method is provided for preventing, reducing or treating plaques and tangles in the brain of an elderly dog ​​or cat, comprising treating the plaques and tangles with any one of the compositions described herein.

[0040] In another aspect, there is provided a method for improving cognitive performance and / or slowing cognitive decline in elderly dogs or cats affected by the formation, deposition, accumulation and / or persistence of cerebral plaques and tangles, comprising administering to the elderly dog ​​or cat any one of the compositions described herein.

[0041] In another aspect, provided is a method for improving athletic performance in a subject by inhibiting or reducing tangles in the brain, the method comprising administering to the subject any one of the compositions described herein. [Brief explanation of the drawings]

[0042] [Figure 1] 1 shows graphs of Aβ1-42 aggregation measured by Thioflavin T fluorometry and Congo Red binding after incubation with Oolong Tea Extract (LOTE) and PTI-00703®. [Figure 2A-C] Representative images of Congo Red, Thioflavin S, and electron micrographs of Aβ fibrils + / - LOTE and PTI-00703® are shown. [Figure 3] Graphs of Aβ1-40 aggregation measured by Thioflavin T fluorometry and Congo Red binding after incubation with LOTE and PTI-00703® are shown. [Figure 4] Circular dichroism spectra of Aβ1-40 after treatment with increasing concentrations of LOTE and PTI-00703® are shown. [Figure 5A-G] 1 depicts data from various internal studies to characterize tau RD fibrils for screening of tau aggregation inhibitors. [Figure 6] 1 shows a graph of tau aggregation measured by thioflavin S fluorometry after incubation with tea extract. [Figure 7] 1 shows a graph of tau aggregation measured by Thioflavin S fluorometry after incubation with LOTE and PTI-00703®. [Figure 8] 1 shows a graph of the secondary structure of tau measured by circular dichroism spectroscopy after treatment with LOTE. [Figure 9] Electron micrographs of tau fibril formation by PTI-00703® and LOTE are shown. [Figure 10] Electron micrographs of pre-formed tau fibrils treated with PTI-00703® and LOTE are shown. [Figure 11] 1 shows a graph of Aβ1-42 aggregation measured by Thioflavin T fluorometry after incubation with Composition 11.1; Composition 11.2; Composition 11.3; Composition 11.4, Composition 11.5, Composition 11.6, and Composition 11.7. [Figure 12]18 shows a graph of Aβ 1-42 aggregation measured by Thioflavin T fluorometry after incubation with Composition 18.1; Composition 18.2; Composition 18.3; Composition 18.4, Composition 18.5, Composition 18.6, and Composition 18.7. [Figure 13] 1 shows a graph of Aβ1-42 aggregation measured by Thioflavin T fluorometry after incubation with Composition 19.1; Composition 19.2; Composition 19.3; Composition 19.5, Composition 19.6, Composition 19.7, Composition 19.8, and Composition 19.9. [Figure 14] 2 shows a graph of Aβ1-42 aggregation measured by Thioflavin T fluorometry after incubation with Composition 21.1; Composition 21.2; Composition 21.3; Composition 21.5; Composition 21.6, and Composition 21.7. [Figure 15] 1 shows cat's claw / oolong tea extract in capsules compared to cat's claw / oolong tea / blackcurrant capsules. [Figure 16] Shown is a glass of water containing a capsule containing cat's claw / oolong tea extract and a capsule containing cat's claw / oolong tea / blackcurrant. [Figures 17A-C] Figure 17A compares the initial dissolution (within 5 seconds) of a single capsule of Cat's Claw / Oolong Tea and a single capsule of Cat's Claw / Oolong Tea / Blackcurrant in water. Figure 17A shows that a single capsule of Cat's Claw / Oolong Tea extract dissolves in water within a few seconds. Figure 17B shows that a single capsule of Cat's Claw / Oolong Tea / Blackcurrant dissolves in water within a few seconds. Note that both solutions initially contain a mass at the top of a glass of water. Figure 17C shows that within 10-15 seconds, the Cat's Claw / Oolong Tea / Blackcurrant combination begins to self-dissolve in water. [Figures 18A-C]The dissolution of cat's claw / oolong tea extract in water is compared with that of cat's claw / oolong tea extract containing blackcurrant. Figure 18A shows the surprising discovery that adding blackcurrant makes the cat's claw / oolong tea extract more water-soluble and tastes much better. Figure 18B shows that the cat's claw / oolong tea extract still contains a lot of water-insoluble material floating on the top of the glass. Figure 18C shows that adding blackcurrant makes the cat's claw / oolong tea extract more water-soluble, with little or no insoluble material at the top of the glass. DETAILED DESCRIPTION OF THE INVENTION

[0043] In one embodiment, the present invention provides a composition comprising blackcurrant extract, Uncaria tomentosa extract, and oolong tea extract, which is useful for treating human subjects with tauopathies, amyloid diseases, brain "plaques and / or tangles," as well as traumatic brain injury (TBI), concussion (observed in most athletes and military / soldiers), post-traumatic stress disorder, head trauma, blows to the head (either single or repeated), and chronic traumatic encephalopathy (CTE). In some embodiments, the compositions effectively inhibit amyloid fibril formation, inhibit amyloid fibril growth, and / or cause the dissolution and / or destruction of pre-formed amyloid fibrils. In some embodiments, the compositions inhibit tau fibril formation (important for subjects with early to mid-stage tauopathy), inhibit tau fibril growth (important for subjects with early to mid-stage tauopathy), and / or cause the dissolution / destruction of pre-formed tau fibrils (important for subjects with late-stage tauopathy). In some embodiments, the compositions are useful for developing cognitive and memory supplements to prevent, reduce, and / or remove brain "plaques and tangles" in humans.

[0044] The present disclosure recognizes that the amount of proanthocyanidins, anthocyanidins, anthocyanins, or combinations thereof present in a blackcurrant extract is important for the activities described herein. In some embodiments, the blackcurrant extract contains about 10% to about 35% w / w of proanthocyanidins, anthocyanidins, anthocyanins, or combinations thereof, preferably about 25% w / w of proanthocyanidins, anthocyanidins, anthocyanins, or combinations thereof. The disclosure also recognizes that various plant extracts, even those derived from the same plant source, have different effects for dissolving plaque and tangles, such as oolong tea extract. In some embodiments, the oolong tea extract is LOTE. In some embodiments, the oolong tea extract is MemorTea®. In some embodiments, the oolong tea extract is obtained from the Guan Yin Mountains in China and provided by AuNutra Industries (Chino, CA). In some embodiments, a composition comprising a blackcurrant extract, an Uncaria tomentosa extract, and an oolong tea extract provides surprising and unexpected activity compared to other plant extracts or each extract alone. In some embodiments, the combination of blackcurrant extract, Uncaria tomentosa extract, and oolong tea extract provides synergistic or complementary efficacy. definition

[0045] Unless otherwise defined, all technical and scientific terms used herein have the meanings commonly understood by those skilled in the art to which this disclosure belongs. As used herein, the following terms have the meanings ascribed to them below, unless otherwise specified. The terms used herein are for the purpose of describing particular embodiments only and are not intended to limit the disclosure.

[0046] As used herein, the singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0047] As used herein, the term "about," when used to modify a numerical value or numerical range, indicates a deviation of 5% to 10% above and 5% to 10% below the value or range while remaining within the intended meaning of the stated value or range.

[0048] As used herein, " control " refers to the alternative sample used in an experiment for comparison purposes. Control can be " positive " or " negative ". For example, when the purpose of an experiment is to determine the correlation between the effectiveness of a therapeutic agent for treating a specific type of disease, a positive control (a composition known to exhibit desired therapeutic effects) and a negative control (a subject or sample that does not receive treatment or receives a placebo) are typically used.

[0049] "Comprising" shall mean that the methods and compositions include the recited elements, but do not exclude others. When used to define methods and compositions, "consisting essentially of" shall mean excluding any other elements that are essential to the combination for the stated purpose. Thus, a composition consisting essentially of the elements defined herein does not exclude trace contaminants from isolation and purification methods, as well as pharmaceutically acceptable carriers such as phosphate-buffered saline, preservatives, etc. "Consisting of" shall mean excluding nothing other than trace elements of other components and substantial method steps for administering the compositions of the invention or process steps for producing the compositions or achieving the intended result. Embodiments defined by each of these transition terms and phrases are within the scope of the present invention.

[0050] A "therapeutically effective amount" generally refers to an amount sufficient to affect the desired degree of disease treatment when administered to a subject or animal for treating a disease. A "therapeutically effective amount" or "therapeutically effective dosage" preferably inhibits, reduces, disrupts, disassembles, or treats the formation, deposition, accumulation, and / or persistence of tau fibrils by at least 20%, more preferably at least 40%, even more preferably at least 60%, and even more preferably at least 80% of the disease associated with these conditions, such as tauopathies, compared to untreated subjects. An effective amount of a compound of the present invention or a composition thereof for treating a mammalian subject is about 0.1 to about 1000 mg / kg / day of the subject's body weight, e.g., about 1 to about 100 mg / kg / day, particularly about 10 to about 100 mg / kg / day. A wide range of dosages of the disclosed compositions are believed to be safe and effective.

[0051] "Anthocyanidins" are the sugar-free counterparts of anthocyanins. Examples of anthocyanidins include, but are not limited to, cyanidin, delphinidin, pelargonidin, peonidin, malvidin, and petunidin.

[0052] "Anthocyanin" is a colored water-soluble pigment belonging to the phenolic group. Anthocyanins are based on flavylium ions or 2-phenylchromenylium ions. Examples of anthocyanins include, but are not limited to, glycosides of cyanidin, delphinidin, malvidin, pelargonidin, peonidin, and petunidin. Examples include, but are not limited to, delphinidin-3-O-glucoside, delphinidin-3-O-rutinoside, cyanidin-3-O-glucoside, and cyanidin-3-O-rutinoside.

[0053] "Amyloid diseases" or "amyloidoses" suitable for treatment with the compositions of the present invention are diseases associated with the formation, deposition, accumulation, and / or persistence of amyloid fibrils, particularly fibrils of amyloid proteins, such as Aβ and IAPP amyloid, selected from the group consisting of beta amyloid protein or Aβ, AA amyloid, AL amyloid, IAPP amyloid, PrP amyloid, α2-microglobulin amyloid, transthyretin, prealbumin, and procalcitonin. Suitable such diseases include Alzheimer's disease, Down's syndrome, mild cognitive impairment (MCI), cognitive impairment disorder (CDD), traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE), concussion, head trauma, single and multiple blows to the head, post-traumatic stress disorder, dementia pugilistica, multiple system atrophy, inclusion body myositis, hereditary cerebral hemorrhage with amyloidosis of the Dutch type, Niemann-Pick disease type C, cerebral beta-amyloid angiopathy, dementia associated with corticobasal degeneration, and type 2 diabetes. amyloidosis of the liver, amyloidosis of chronic inflammation, amyloidosis of malignancy and familial Mediterranean fever, amyloidosis of multiple myeloma and B-cell cachexia, amyloidosis of prion diseases, amyloidosis associated with Creutzfeldt-Jakob disease, Gerstmann-Straussler syndrome, kuru, scrapie, carpal tunnel syndrome, senile cardiac amyloidosis, familial amyloidotic polyneuropathy, and amyloidosis associated with endocrine tumors.

[0054] "Fibrillogenesis" refers to the formation, deposition, accumulation and / or persistence of tau fibrils, filaments, inclusions, deposits, inclusion bodies, and the like.

[0055] "Inhibition of fibril formation" refers to the inhibition of the formation, deposition, accumulation and / or persistence of such amyloid "plaques" or tau "tangle" fibril-like deposits.

[0056] "Disruption of fibril or fibril formation" refers to the disruption of preformed beta-amyloid or tau fibrils, which typically exist in a predominant beta-sheet secondary structure. Such disruption by the compounds of the invention can include a significant reduction or disassembly of beta-amyloid or tau fibrils, as assessed by various methods, such as circular dichroism spectroscopy, thioflavin S fluorometry, SDS-PAGE / Western blotting, or negative stain electron microscopy, as demonstrated by the examples presented in this application.

[0057] "Mammal" includes both human and non-human mammals, such as companion animals (dogs, cats, etc.), laboratory animals (mice, rats, guinea pigs, etc.), and farm animals (cows, horses, sheep, goats, pigs, etc.). In some embodiments, the mammal is a human. In some embodiments, the mammal is a dog or a cat.

[0058] "Plaques" refer to meatball-like "amyloid deposits" composed of beta-amyloid protein, or Aβ, found in various brain regions (e.g., hippocampus, cerebral cortex, frontal lobe). They are a pathological hallmark of brain aging, mild cognitive impairment (MCI), and Alzheimer's disease, and are found in aging mammals such as dogs, cats (called cognitive canine dysfunction, or CDD), monkeys, polar bears, and horses. The accumulation of amyloid "plaques" in the brain is thought to lead to neurodegeneration, loss of synapses and interneuronal connections, cognitive decline, memory decline and loss, and loss of focus and concentration.

[0059] "Proanthocyanidins" are oligomeric flavonoids. Proanthocyanidins include dimers or oligomers of catechin and epicatechin, and their gallate esters.

[0060] The term "subject," as used herein, refers to an animal that is the object of treatment, observation, or experiment. By way of example, a subject may be a mammal, such as, but not limited to, a human, a cat, or a dog. In some embodiments, the subject is a human. In some embodiments, the human subject is a patient. In some embodiments, the subject is a cat. In some embodiments, the subject is a dog.

[0061] "Tangles" refers to "dried," spaghetti-like "tangle deposits" of tau protein found in various regions of the brain (e.g., hippocampus, cerebral cortex, frontal cortex), which are a pathological hallmark of brain aging, mild cognitive impairment, Alzheimer's disease, concussion, traumatic brain injury (TBI), single and repeated blows to the head, post-traumatic stress disorder, and chronic traumatic encephalopathy (CTE).

[0062] "Tauopathies" suitable for treatment with the compounds of the present invention are also diseases associated with the formation, deposition, accumulation, or persistence of tau fibrils. Suitable diseases include Alzheimer's disease, frontotemporal lobar degeneration with tau inclusions (FTLD-tau), e.g., Pick's disease, progressive supranuclear palsy, and corticobasal degeneration, argyrophilic grain dementia, some prion diseases, amyotrophic lateral sclerosis / parkinsonism-dementia complex of Guam (also known as Richkova-Dig's disease), dementia pugilistica, chronic traumatic encephalopathy, Parkinson's disease, especially some genetic forms of Parkinson's disease, and tangle-predominant dementia (with neurofibrillary tangles similar to those of Alzheimer's disease but without amyloid plaques).

[0063] Tau fibrils is a general term referring to a group of diverse but specific intra- or extracellular protein deposits that all have common morphological characteristics, staining properties, and X-ray diffraction spectra.

[0064] "Treating" a disease or "treatment" of a disease includes preventing the disease from occurring in a mammal susceptible to the disease but that has not yet experienced or exhibited symptoms of the disease (prophylactic treatment). Treating can also mean inhibiting the disease (slowing or halting its development), providing relief from symptoms or side effects of the disease (including palliative treatment), or alleviating the disease (causing regression of the disease), such as by disrupting preformed tau fibrils. Treatment need not be complete. One such preventative treatment can be the use of the disclosed compounds for the treatment of mild cognitive impairment (MCI). composition

[0065] In one aspect, a composition is provided that includes a therapeutically effective amount of a blackcurrant extract, a therapeutically effective amount of an Uncaria tomentosa extract, and a therapeutically effective amount of an oolong tea extract.

[0066] In one aspect, a composition is provided comprising a therapeutically effective amount of a blackcurrant extract, a therapeutically effective amount of an Uncaria tomentosa extract, a therapeutically effective amount of an oolong tea extract, and one or more pharmaceutically acceptable excipients or carriers.

[0067] In one aspect, a composition is provided consisting essentially of a therapeutically effective amount of blackcurrant extract, a therapeutically effective amount of Uncaria tomentosa extract, a therapeutically effective amount of oolong tea extract, and one or more pharmaceutically acceptable excipients or carriers.

[0068] In another aspect, a composition is provided comprising a therapeutically effective amount of blackcurrant extract. In some embodiments, the composition further comprises a therapeutically effective amount of Uncaria tomentosa extract. In some embodiments, the composition further comprises a therapeutically effective amount of oolong tea extract. In some embodiments, the composition further comprises a therapeutically effective amount of Uncaria tomentosa extract and a therapeutically effective amount of oolong tea extract.

[0069] In some embodiments, the compositions described herein benefit human subjects with amyloidosis and elderly mammals (such as dogs and cats, which develop amyloid "plaques" in the brain with age). In some embodiments, the compositions described herein are useful for treating / inhibiting amyloid deposition, accumulation, and / or persistence in amyloidosis. In some embodiments, the compositions described herein are useful for inhibiting the formation, deposition, accumulation, and / or persistence of tau fibrils.

[0070] In some embodiments, a composition comprising the extracts described herein inhibits tau fibril formation with only one of the extracts and has the ability to rapidly disaggregate preformed tau fibrils. In some embodiments, a composition comprising the extracts described herein inhibits amyloid fibril formation with only one of the extracts and has the ability to rapidly disaggregate preformed amyloid fibrils.

[0071] In some embodiments, the fibril inhibitory activity or efficacy of the compositions described herein is greater than about 50%, greater than about 55%, greater than about 60%, greater than about 65%, greater than about 70%, greater than about 75%, greater than about 80%, greater than about 85%, greater than about 90%, greater than about 95%, or greater than about 99%.

[0072] In some embodiments, the composition is formulated in a single dosage form, such as a capsule. In some embodiments, the capsule is about 100 mg to about 2000 mg. In some embodiments, the capsule is about 100 mg to about 1000 mg. In some embodiments, the capsule is about 200 mg to about 2000 mg. In some embodiments, the capsule is about 200 mg to about 1000 mg. In some embodiments, the capsules contain about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg, about 500 mg, about 525 mg, about 550 mg, about 575 mg, about 600 mg, about 625 mg, about 650 mg, about 670 mg, about 675 mg, about 700 mg, about 750 ... mg, about 725 mg, about 750 mg, about 775 mg, about 800 mg, about 825 mg, about 850 mg, about 875 mg, about 900 mg, about 925 mg, about 950 mg, about 975 mg, about 1000 mg, about 1050 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, about 1500 mg, about 1600 mg, about 1700 mg, about 1800 mg, about 1900 mg, or about 2000 mg. Blackcurrant

[0073] Grown for its berries, blackcurrant (Ribes nigrum) is a woody shrub in the Ribes family (Grossulariaceae). It is native to temperate regions of Central and Northern Europe, New Zealand, and Northern Asia and is widely cultivated commercially and domestically. In addition to its high vitamin C content, the berry contains flavonoids and at least 15 phenolic acids, including anthocyanins, anthocyanidins, and proanthocyanidins. The major anthocyanins identified in blackcurrant include delphinidin-3-O-glucoside, delphinidin-3-O-rutinoside, cyanidin-3-O-glucoside, and cyanidin-3-O-rutinoside.

[0074] This disclosure recognizes that the amount of anthocyanins, anthocyanidins, proanthocyanidins, or combinations thereof present in a blackcurrant extract is important for activity. In some embodiments, the blackcurrant extract contains about 10% to about 35% w / w of proanthocyanidins, anthocyanidins, anthocyanins, or combinations thereof. In some embodiments, the blackcurrant extract contains greater than 15% w / w of proanthocyanidins, anthocyanidins, anthocyanins, or combinations thereof. In some embodiments, the blackcurrant extract contains about 15% to about 35% w / w of proanthocyanidins, anthocyanidins, anthocyanins, or combinations thereof. In some embodiments, the blackcurrant extract contains about 15% to about 30%, about 15% to about 25%, about 15% to 20%, about 20% to about 35%, and about 20% to about 30% w / w of any of proanthocyanidins, anthocyanidins, anthocyanins, or combinations thereof. In some embodiments, the blackcurrant extract comprises about 10% w / w, about 11% w / w, about 12% w / w, about 13% w / w, about 14% w / w, about 15% w / w, about 16% w / w, about 17% w / w, about 18% w / w, about 19% w / w, about 20% w / w, about 21% w / w, about 22% w / w, about 23% w / w, about 24% w / w, about 25% w / w, about 26% w / w, about 27% w / w, about 28% w / w, about 29% w / w, about 30% w / w, about 31% w / w, about 32% w / w, about 33% w / w, about 34%, or about 35% w / w of proanthocyanidins, anthocyanidins, anthocyanins, or combinations thereof. In some embodiments, the blackcurrant extract comprises about 25% w / w proanthocyanidins, anthocyanidins, anthocyanins, or combinations thereof. In some embodiments, the blackcurrant extract comprises about 20% to about 30% w / w proanthocyanidins, anthocyanidins, anthocyanins, or combinations thereof. In some embodiments, the blackcurrant extract comprises about 15% to about 35% w / w proanthocyanidins, anthocyanidins, anthocyanins, or combinations thereof.

[0075] In some embodiments, the blackcurrant extract comprises anthocyanins, anthocyanidins, proanthocyanidins, or combinations thereof. In some embodiments, the blackcurrant extract comprises anthocyanins. In some embodiments, the blackcurrant extract comprises anthocyanidins. In some embodiments, the blackcurrant extract comprises proanthocyanidins. In some embodiments, the blackcurrant extract comprises a combination of anthocyanins, anthocyanidins, and / or proanthocyanidins.

[0076] In some embodiments, the composition comprises about 10 mg to about 500 mg of blackcurrant extract. In some embodiments, the composition comprises about 10 mg to about 250 mg of blackcurrant extract. In some embodiments, the composition comprises about 10 mg to about 100 mg of blackcurrant extract. In some embodiments, the composition comprises about 30 mg to about 100 mg of blackcurrant extract. In some embodiments, the composition comprises about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, or about 500 mg of blackcurrant extract.

[0077] In some embodiments, the composition comprises from about 0.1% to about 30% w / w of blackcurrant extract. In some embodiments, the composition comprises from about 0.1% to about 20% w / w of blackcurrant extract. In some embodiments, the composition comprises from about 0.1% to about 10% w / w of blackcurrant extract. In some embodiments, the composition comprises from about 1% to about 30% w / w of blackcurrant extract. In some embodiments, the composition comprises from about 1% to about 20% w / w of blackcurrant extract. In some embodiments, the composition comprises from about 1% to about 10% w / w of blackcurrant extract. In some embodiments, the composition comprises about 0.1% w / w, about 0.2% w / w, about 0.3% w / w, about 0.4% w / w, about 0.5% w / w, about 0.6% w / w, about 0.7% w / w, about 0.8% w / w, about 0.9% w / w, about 1% w / w, about 2% w / w, about 3% w / w, about 4% w / w, about 5% w / w, about 6% w / w, about 7% w / w, about 8% w / w, about 9% w / w, about 10% w / w, about 11% w / w, about 12% w / w, about 13% w / w, about 14% w / w, about 15% w / w, about 16% w / w, about 17% w / w, about 18% w / w, about 19% w / w, about 20% w / w, about 21% w / w, about 22% w / w, about 23% w / w, about 24% w / w, about 25% w / w, about 26% w / w, about 27% w / w, about 28% w / w, about 29% w / w, or about 30% w / w of blackcurrant extract. Cat's Claw

[0078] The plant Uncaria tomentosa, also known as "Unade Gato" (Spanish) or "Cat's claw" (English), refers to a woody vine that grows in the Amazon rainforest. Other names for cat's claw include paraguayo, garabato, garbato casha, tambor huasca, una de gavilan, hawk's claw, nail of cat, and nail of cat schuler. This slow-growing vine takes 20 years to reach maturity and can grow to over 100 feet in length as it attaches and wraps around native trees. It is found abundantly in the foothills at elevations between 2 and 8,000 feet. This vine is called "cat's claw" because of its distinctive curved, claw-like thorns that protrude from the base of its leaves. Uncaria tomentosa is expected to have immune-supporting, anti-inflammatory, antiviral, antimutagenic, and antioxidant properties. Its anti-inflammatory properties are expected to be beneficial in treating, for example, arthritis, rheumatism, bursitis, and gout. Without being bound by theory, it is believed that its beneficial effects in treating arthritic pain may be due, in part, to its ability to cleanse the digestive tract and help remove toxins from the body. Furthermore, Uncaria tomentosa, or cat's claw, is expected to alleviate pain, for example, associated with chemotherapy, radiation therapy, and AZT use.

[0079] Uncaria tomentosa, or cat's claw, is expected to help stop viral infections at an early stage, fight opportunistic infections in AIDS patients, and reduce the viable size of some skin tumors and cysts. Uncaria tomentosa can also be used to treat a variety of illnesses, including cancer, AIDS, Crohn's disease, respiratory infections, allergies, herpes, prostate problems, lupus, Epstein-Barr virus, chronic fatigue syndrome, and various stomach and intestinal disorders.

[0080] For additional details and background regarding Uncariatomentosa, see also our WIPO International Publication No. International Application No. 98 / 51302, which is incorporated herein by reference in its entirety.

[0081] The present inventors previously discovered and disclosed a naturally occurring plant product, the inner bark and / or root from the plant Uncaria tomentosa, or Cat's Claw (PTI-00703®), in WIPO Publication No. WO 98 / 51302, entitled "Composition and Methods for Treating Alzheimer's Disease and other Amyloidoses," dated November 19, 1998, which is incorporated herein by reference in its entirety. As disclosed herein, this plant compound alone has surprising efficacy in disrupting and / or dissolving amyloid deposits and other buildups, and is believed to be a potent inhibitor of amyloid formation in Alzheimer's disease, type 2 diabetes, and other amyloidoses.

[0082] Uncaria tomentosa extract PTI-00703® is also referred to as "703" or "PTI-703" for the therapeutic intervention of amyloidosis. PTI-00703® is derived from the bark of the Cat's Claw tree, found in the Amazon rainforest. In some embodiments, the Uncaria tomentosa extract is PTI-00703. In some embodiments, the Uncaria tomentosa extract is derived from the inner bark and / or root. In some embodiments, the Uncaria tomentosa extract is derived from the inner bark or root of the Amazon rainforest.

[0083] In some embodiments, the composition comprises about 100 mg to about 500 mg of Uncaria tomentosa extract. In some embodiments, the composition comprises about 100 mg to about 300 mg of Uncaria tomentosa extract. In some embodiments, the composition comprises about 100 mg to about 250 mg of Uncaria tomentosa extract. In some embodiments, the composition comprises about 100 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about Contains 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, or about 500 mg of Uncaria tomentosa extract.

[0084] In some embodiments, the composition comprises about 5% to about 60% w / w of Uncaria tomentosa extract. In some embodiments, the composition comprises about 5% to about 40% w / w of Uncaria tomentosa extract. In some embodiments, the composition comprises about 10% to about 60% w / w of Uncaria tomentosa extract. In some embodiments, the composition comprises about 10% to about 40% w / w of Uncaria tomentosa extract. In some embodiments, the composition comprises about 10% w / w, about 11% w / w, about 12% w / w, about 13% w / w, about 14% w / w, about 15% w / w, about 16% w / w, about 17% w / w, about 18% w / w, about 19% w / w, about 20% w / w, about 21% w / w, about 22% w / w, about 23% w / w, about 24% w / w, about 25% w / w, about 26% w / w, about 27% w / w, about 28% w / w, about 29% w / w, about 30% w / w, about 31% w / w, about 32% w / w, about 33% w / w, about 34% w / w, about 35% w / w. / w, about 36% w / w, about 37% w / w, about 38% w / w, about 39% w / w, about 40% w / w, about 41% w / w, about 42% w / w, about 43% w / w, about 44% w / w, about 45% w / w, about 46% w / w, about 47% w / w, about 48% w / w, about 49% w / w, about 50% w / w, about 51% w / w, about 52% w / w, about 53% w / w, about 54% w / w, about 55% w / w, about 56% w / w, about 57% w / w, about 58% w / w, about 59% w / w, or about 60% w / w of Uncaria tomentosa extract. oolong tea

[0085] Oolong tea is a Chinese tea (Camellia sinensis) produced through a process that involves wilting and oxidizing the plant under intense sunlight, followed by curling and twisting. The inventors have screened over 25 tea extracts, along with coffee and yerba mate extracts, for in vitro amyloid fibril aggregation inhibition and disaggregation using various assays. The inventors have recognized that the use of oolong tea extract in combination with Uncaria tomentosa extract exhibits amyloid fibril aggregation inhibition and disaggregation. In some embodiments, the oolong tea extract is LOTE. In some embodiments, the oolong tea extract is MemorTea®. In some embodiments, the oolong tea extract is obtained from the Guan Yin Mountains in China and provided by AuNutra Industries (Chino, CA).

[0086] In some embodiments, the composition comprises about 100 mg to about 500 mg of oolong tea extract. In some embodiments, the composition comprises about 100 mg to about 250 mg of oolong tea extract. In some embodiments, the composition comprises about 100 mg to about 300 mg of oolong tea extract. In some embodiments, the composition comprises about 100 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, or about 350 mg of oolong tea extract. mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, about 500 mg of oolong tea extract.

[0087] In some embodiments, the composition comprises about 5% to about 60% w / w oolong tea extract. In some embodiments, the composition comprises about 5% to about 40% w / w oolong tea extract. In some embodiments, the composition comprises about 10% to about 60% w / w oolong tea extract. In some embodiments, the composition comprises about 10% to about 40% w / w oolong tea extract. In some embodiments, the composition comprises about 10% w / w, about 11% w / w, about 12% w / w, about 13% w / w, about 14% w / w, about 15% w / w, about 16% w / w, about 17% w / w, about 18% w / w, about 19% w / w, about 20% w / w, about 21% w / w, about 22% w / w, about 23% w / w, about 24% w / w, about 25% w / w, about 26% w / w, about 27% w / w, about 28% w / w, about 29% w / w, about 30% w / w, about 31% w / w, about 32% w / w, about 33% w / w, about 34% w / w, or about The composition comprises 35% w / w, about 36% w / w, about 37% w / w, about 38% w / w, about 39% w / w, about 40% w / w, about 41% w / w, about 42% w / w, about 43% w / w, about 44% w / w, about 45% w / w, about 46% w / w, about 47% w / w, about 48% w / w, about 49% w / w, about 50% w / w, about 51% w / w, about 52% w / w, about 53% w / w, about 54% w / w, about 55% w / w, about 56% w / w, about 57% w / w, about 58% w / w, about 59%, or about 60% w / w of oolong tea extract.

[0088] In some exemplary embodiments, the composition comprises 1% to 20% w / w of blackcurrant extract containing 25% anthocyanins, 10% to 50% w / w of Uncaria tomentosa extract, and 10% to 50% w / w of oolong tea extract. In some exemplary embodiments, the composition comprises 1% to 15% w / w of blackcurrant extract containing 25% anthocyanins, 10% to 45% w / w of Uncaria tomentosa extract, and 10% to 45% w / w of oolong tea extract. In some exemplary embodiments, the composition comprises 1% to 10% w / w of blackcurrant extract containing 25% anthocyanins, 10% to 40% w / w of Uncaria tomentosa extract, and 10% to 40% w / w of oolong tea extract. In some exemplary embodiments, the composition comprises 1% to 10% w / w blackcurrant extract containing 25% anthocyanins, 20% to 40% w / w Uncaria tomentosa extract, and 20% to 40% w / w oolong tea extract.

[0089] In some exemplary embodiments, the composition comprises blackcurrant extract, Uncaria tomentosa extract, and oolong tea extract, wherein the blackcurrant extract, Uncaria tomentosa extract, and oolong tea extract are active ingredients. In some embodiments, the blackcurrant extract comprises about 10% to about 35% w / w of proanthocyanidins, anthocyanidins, anthocyanins, or combinations thereof. In some embodiments, the blackcurrant extract comprises about 25% w / w of proanthocyanidins, anthocyanidins, anthocyanins, or combinations thereof. In some embodiments, the blackcurrant is about 1% to 20% w / w, 1% to 15% w / w, or 1% to 10% w / w of the active ingredient of the composition, wherein the blackcurrant extract, Uncaria tomentosa extract, and oolong tea extract are active ingredients of the composition. In some embodiments, Uncaria tomentosa extract is about 10%-50% w / w, 10%-45% w / w, 10%-40% w / w, or about 20%-40% w / w of the active ingredient of the composition, and blackcurrant extract, Uncaria tomentosa extract, and oolong tea extract are active ingredients of the composition. In some embodiments, oolong tea extract is about 10%-50% w / w, 10%-45% w / w, 10%-40% w / w, or about 20%-40% w / w of the active ingredient of the composition, and blackcurrant extract, Uncaria tomentosa extract, and oolong tea extract are active ingredients of the composition.

[0090] In one aspect, a blended composition comprising blackcurrant extract, Uncaria tomentosa extract, and oolong tea extract is more soluble (e.g., water-soluble) than a blended composition without blackcurrant extract, such as a composition comprising Uncaria tomentosa extract and oolong tea extract as active agents. A blended composition comprising blackcurrant extract, Uncaria tomentosa extract, and oolong tea extract is more soluble (e.g., water-soluble) than a blended composition without blackcurrant extract, such as a composition consisting of Uncaria tomentosa extract and oolong tea extract. In some embodiments, the addition of blackcurrant extract to a composition comprising Uncaria tomentosa extract and oolong tea extract increases the water solubility of Uncaria tomentosa extract and oolong tea extract. In some embodiments, the addition of blackcurrant extract to a composition comprising Uncaria tomentosa extract and oolong tea extract increases the solubility (e.g., water solubility or water solubility) of the Uncaria tomentosa extract and / or oolong tea extract by at least 1-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 30-fold, at least 40-fold, or at least 50-fold, e.g., 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold. In some embodiments, the addition of blackcurrant extract to a composition comprising Uncaria tomentosa extract and oolong tea extract increases the solubility (e.g., water solubility or water solubility) of the Uncaria tomentosa extract and / or oolong tea extract by at least 1-fold to at least 50-fold, or by at least 1-fold to at least 20-fold, e.g., 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold.In some embodiments, the addition of blackcurrant extract to a composition comprising Uncaria tomentosa extract and oolong tea extract increases the solubility (e.g., water solubility or water solubility) of the Uncaria tomentosa extract and / or oolong tea extract by at least 1-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 30-fold, at least 40-fold, or at least 50-fold, e.g., 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold, over a composition that does not contain blackcurrant extract, such as a composition comprising Uncaria tomentosa extract and oolong tea extract as the only active agents. In some embodiments, a composition comprising blackcurrant extract, Uncaria tomentosa extract, and oolong tea extract has increased solubility (e.g., water solubility or water solubility) by at least 1-fold to at least 50-fold, or by at least 1-fold to at least 20-fold, e.g., about 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold, over a composition that does not contain blackcurrant extract, such as a composition comprising Uncaria tomentosa extract and oolong tea extract as the only active agents. In some embodiments, solubility (e.g., water solubility or water solubility) is increased by any of the following: 1x to 50x, 5x to 50x, 5x to 40x, 5x to 30x, 5x to 20x, 10x to 50x, 10x to 40x, 10x to 35x, 10x to 30x, 10x to 25x, and 10x to 20x. This is a surprising discovery: Uncaria tomentosa extract and oolong tea extract (particularly Uncaria tomentosa extract) are highly water-insoluble. The addition of blackcurrant extract as a third ingredient surprisingly makes the resulting composition containing all three ingredients highly water-soluble, resulting in a much more pleasant taste (less astringent and bitter). Making the ingredients more water-soluble through the addition of blackcurrant also allows the resulting product to be used as a beverage, a drinking shot, and / or a product for people who cannot swallow pills, caplets, tablets, capsules, etc. use

[0091] In one embodiment, the compositions described herein are useful for the development of cognitive and memory supplements to inhibit, prevent, reduce, and / or remove brain "plaques and / or tangles" in humans.

[0092] The compositions described herein also benefit elderly dogs and cats that develop brain "plaques" (and to a lesser extent "tangles") and have dementia as defined in Canine Cognitive Disorder (CCD).

[0093] In another aspect, provided herein is a pet food supplement for geriatric dogs that reduces amyloid "plaques" in the brain and improves cognition, memory, short-term memory, concentration, and attention span.

[0094] In another aspect, provided herein is a pet food supplement for senior cats that inhibits, prevents, and / or reduces amyloid "plaques" in the brain and improves cognition, memory, short-term memory, concentration, and attention span.

[0095] In another aspect, the compositions described herein are used to treat the formation, deposition, accumulation, and / or persistence of tau fibrils, such as those occurring in tauopathies. In some embodiments, the compositions described herein are used to treat tauopathies, for example, to prevent and / or treat traumatic brain injury (TBI), concussion (observed in most athletes and military / soldiers), post-traumatic stress disorder, head trauma, blows to the head (single or repeated), and chronic traumatic encephalopathy (CTE).

[0096] In another aspect, provided herein are compositions that are effective inhibitors and / or reducers of brain "tangles," such as those seen in people with traumatic brain injury (TBI), concussion (seen in most athletes and military / soldiers), post-traumatic stress disorder, head trauma, blows to the head (single or repeated), and chronic traumatic encephalopathy (CTE).

[0097] In another aspect, provided herein are compositions that effectively inhibit and / or prevent amyloid fibril formation, and / or prevent inhibition of amyloid fibril growth, and / or cause dissolution and / or destruction of pre-formed amyloid fibrils.

[0098] In another aspect, provided herein are compositions that prevent and / or inhibit the formation of tau fibrils (important for subjects with early to mid-stage tauopathy), inhibit the growth of tau fibrils (important for subjects with early to mid-stage tauopathy), and / or cause the dissolution / destruction of pre-formed tau fibrils (important for subjects with late-stage tauopathy).

[0099] In another aspect, the compositions described herein inhibit tau fibril formation, inhibit tau fibril growth, inhibit tau fibril-proteoglycan interactions, inhibit tau fibril-proteoglycan and / or glycosaminoglycan interactions, and cause dissolution and / or disruption of pre-formed tau fibrils. method

[0100] In another aspect, a method of treating amyloidosis in a mammal in need thereof is provided, comprising administering to a mammal in need thereof any one of the compositions described herein. In some embodiments, the amyloidosis is Alzheimer's disease, Down's syndrome, dementia pugilistica, cognitive impairment syndrome, canine cognitive impairment, multiple system atrophy, inclusion body myositis, hereditary cerebral hemorrhage with Dutch amyloidosis, Niemann-Pick disease type C, cerebral amyloid angiopathy, dementia associated with corticobasal degeneration, amyloidosis in type 2 diabetes, amyloidosis in chronic inflammation, amyloidosis in malignancy and familial Mediterranean fever, amyloidosis in multiple myeloma and B-cell dyscrasia, amyloidosis in prion disease, Creutzfeldt-Jakob disease, Gerstmann-Straussler syndrome, amyloidosis associated with kuru, scrapie, carpal tunnel syndrome, senile cardiac amyloidosis, familial amyloidotic polyneuropathy, or amyloidosis associated with endocrine tumors.

[0101] Also provided in one embodiment is a method of treating amyloid fibril formation, deposition, accumulation, or persistence, comprising treating the fibrils with any one of the compositions described herein.

[0102] In another aspect, a method of treating the formation, deposition, accumulation, or persistence of beta-amyloid-containing plaques is provided, comprising treating the plaques with any one of the compositions described herein.

[0103] In another aspect, there is provided a method for the treatment of a tauopathy in a human or mammal in need thereof, comprising administering to a human or mammal in need thereof any one of the compositions described herein. In some embodiments, the tauopathy is Alzheimer's disease, frontotemporal lobar degeneration with inclusion bodies (FLTD-tau), Pick's disease, progressive supranuclear palsy, corticobasal degeneration, argyrophilic grain dementia, prion disease, amyotrophic lateral sclerosis-parkinsonism-dementia complex of Guam (also known as Richkovaig's disease), Parkinson's disease, tangle-predominant dementia, ganglion, ganglioglioma, gangliocytoma, meningiomatosis, subacute sclerosing panencephalitis, lead encephalopathy, tuberous sclerosis, Hallervorden-Spatz disease, lipofuscinosis, traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE), dementia pugilistica, concussion, single or repeated blows to the head, or post-traumatic stress disorder.

[0104] In one aspect, a method is provided for treating the formation, deposition, accumulation, or persistence of tangles comprising tau protein, comprising treating the tangles with any one of the compositions described herein.

[0105] In one aspect, provided is a method for improving cognitive performance and / or slowing cognitive decline in a subject in need thereof, comprising administering to the subject any one of the compositions described herein to reduce the formation, deposition, accumulation or persistence of beta-amyloid protein plaques or fibrils in the subject.

[0106] In one aspect, provided is a method for improving cognitive performance and / or slowing cognitive decline in a subject suffering from an amyloid disease, comprising administering to the subject any one of the compositions described herein to reduce the formation, deposition, accumulation or persistence of beta-amyloid protein plaques or fibrils in the subject.

[0107] In one aspect, provided is a method for improving learning, memory, cognition, concentration and / or attention in a subject in need thereof, comprising administering to the subject any one of the compositions described herein.

[0108] In one aspect, provided is a method for improving learning, memory, cognition, concentration and / or attention in a subject with age-associated memory impairment (AAMI), mild cognitive impairment (MCI), or Alzheimer's disease, the method comprising administering to the subject any one of the compositions described herein.

[0109] In one aspect, provided is a method for preventing, reducing, or treating tangles in the brain of a subject in need thereof, the method comprising administering to the subject any one of the compositions described herein.

[0110] In one aspect, provided is a method for preventing, reducing, or treating tangles in the brain of a subject who has experienced single or multiple concussions, traumatic brain injury (TBI), blows to the head, chronic traumatic encephalopathy (CTE), post-traumatic stress disorder, brain aging, mild cognitive impairment, or Alzheimer's disease, comprising administering to the subject any one of the compositions described herein.

[0111] In one aspect, provided is a method for preventing, reducing, or treating plaque in the brain of a subject in need thereof, the method comprising administering to the subject any one of the compositions described herein.

[0112] In one aspect, a method is provided for preventing, reducing or treating plaques and tangles in the brain of an elderly dog ​​or cat, comprising treating the plaques and tangles with any one of the compositions described herein.

[0113] In one aspect, provided is a method for improving cognitive performance and / or slowing cognitive decline in an elderly dog ​​or cat affected by the formation, deposition, accumulation and / or persistence of brain plaques and tangles, comprising administering to the elderly dog ​​or cat any one of the compositions described herein.

[0114] In one aspect, a method is provided for improving athletic performance in a subject by inhibiting or reducing tangles in the brain, comprising administering to the subject any one of the compositions described herein. In some embodiments, the subject is an athlete. Dosage

[0115] In some embodiments, the composition has a therapeutically effective amount of a combination of blackcurrant extract, oolong tea extract, and Uncaria tomentosa extract in a dosage range of about 0.1 to about 500 mg / kg of subject body weight, more preferably about 1.0 to about 100 mg / kg of subject body weight.

[0116] However, it will be understood that the therapeutic dosage administered will be determined by a physician in light of the relevant circumstances, including the clinical condition being treated, the organs or tissues affected or suspected to be affected by tau fibril accumulation, and the selected route of administration. Accordingly, the dosage ranges set forth above are not intended to limit the scope of the invention in any way.

[0117] The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce a desired therapeutic effect, in association with a suitable pharmaceutical carrier. Administration

[0118] Generally, the compositions described herein are administered in any of the usual ways known in the art.Administration can be by any one of the following routes: oral, topical, and systemic (for example, transdermal, intranasal, or suppository), or parenteral (for example, intramuscular, subcutaneous, or intravenous injection).In some embodiments, the compositions are administered orally, such as by oral capsule, drink preparation, drink shot, or any other method, or by aerosol spray, or parenterally injectable or injectable form.

[0119] In some embodiments, the composition is orally administered in the form of a drink shot.As used herein, a drink shot is a drink that contains any one of the compositions described herein in a small amount of liquid (such as about 50 mL).Or in other words, a drink shot is a concentrated drink. compound

[0120] In some embodiments, the blackcurrant extract, Uncaria tomentosa extract, and oolong tea extract are combined in a single dosage form. In some embodiments, any two of the blackcurrant extract, Uncaria tomentosa extract, and oolong tea extract are combined in a single dosage form. In some embodiments, any one of the blackcurrant extract, Uncaria tomentosa extract, and oolong tea extract is combined in a single dosage form.

[0121] The composition may take the form of a tablet, pill, capsule, semisolid, powder, sustained-release formulation, solution, drink, drink shot, suspension, elixir, aerosol, or any other suitable composition, and include at least one pharmaceutically acceptable excipient, carrier, or diluent. Suitable excipients, carriers, and diluents are known to those skilled in the art. Methods for formulating compositions are described in standard references such as Alfonso AR: Remington's Pharmaceutical Sciences, 17th Edition, Mack Publishing Company (Easton, Pa., 1985). Suitable liquid carriers, particularly for injectable solutions, include water, aqueous saline, aqueous dextrose, and glycols.

[0122] In particular, the compound(s) can be orally administered, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, dissolved fizz tablets, emulsions, hard or soft capsules, syrups or elixirs.In one embodiment, only one such compound is administered in any particular dosage form.Compositions intended for oral use can be prepared by any method known in the art for producing dietary supplement compositions, and such compositions may contain one or more agents selected from the group consisting of sweeteners, flavoring agents, coloring agents, and preservatives to provide a nutritious and refreshing preparation.

[0123] In some embodiments, the components of the composition may be commercially available in any form and further adjusted using suitable carriers, excipients, and diluents, including lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water syrup, methylcellulose, methyl and propylhydroxybenzoates, talc, magnesium stearate, and mineral oil. The formulation may additionally contain lubricants, wetting agents, emulsifiers and suspending agents, preservatives, sweeteners, or flavoring agents. The compositions of the present invention may be formulated to provide a rapid, sustained, or delayed response of the active ingredient after administration to a subject.

[0124] The tablets contain the extracts described herein mixed with non-toxic pharmaceutically acceptable excipients suitable for tablet manufacture.These excipients include inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate, granulating and disintegrating agents such as corn starch or alginic acid, binders such as corn starch, gelatin or acacia, and lubricants such as magnesium stearate, stearic acid or talc.The tablets can be uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained effect over a longer period of time.For example, a time-delay material such as glycerol monostearate or glycerol distearate can be used. Formulations for oral use can also be prepared as hard or soft gelatin capsules, in which the compound is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate, or kaolin, and in which the active ingredient is mixed with water or an oil medium such as peanut oil, liquid paraffin, or olive oil.

[0125] Aqueous suspensions contain the compound mixed with excipients suitable for the manufacture of aqueous suspensions.Such excipients include, for example, suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum and acacia gum; dispersing agents and wetting agents, which are naturally occurring phosphatides, such as lecithin, or condensation products of alkylene oxides with fatty acids; such as polyoxyethylene stearate, or condensation products of ethylene oxide with long-chain aliphatic alcohols, such as heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids such as hexitols, such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, such as polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives, for example, ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and / or one or more sweetening agents, such as sucrose or saccharin.

[0126] Oily suspensions can be formulated by suspending the extract in vegetable oils, such as arachis oil, olive oil, sesame oil, or coconut oil, or mineral oils such as liquid paraffin.Oily suspensions can contain thickening agents, such as beeswax, paraffin, or cetyl alcohol.Sweeteners and flavoring agents, as described below, can be added to provide a palatable oral preparation.These compositions can be preserved by adding antioxidants, such as ascorbic acid.

[0127] Dispersible powders and granules suitable for preparing aqueous suspensions by adding water provide the active ingredient mixed with a dispersing or wetting agent, a suspending agent, and one or more preservatives.Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above.Additional excipients, such as sweeteners, flavoring agents, and other additives, may also be present.

[0128] The composition may also be in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil, such as olive oil or arachis oil, or a mineral oil, such as liquid paraffin, or a mixture thereof. Suitable emulsifiers may be naturally occurring gums, such as gum arabic or gum tragacanth, naturally occurring phosphatides, such as soybeans, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate, and condensation products of the above partial esters with ethylene oxide, such as polyethylene sorbitan monooleate. Emulsions may also contain sweeteners and flavoring agents. Syrups and elixirs may be formulated with sweeteners, such as glycerol, sorbitol, or sucrose. Such formulations may also contain analgesics, preservatives, flavorings, and coloring agents.

[0129] The following non-limiting examples are given by way of illustration only and are not to be considered limitations of the present disclosure, many obvious variations of which are possible without departing from its spirit or scope. Example

[0130] The present technology is further illustrated by the following examples, which should not be construed as limiting in any way, and which demonstrate the preparation, characterization, and use of the compositions described herein. Preparation of Composition-Components

[0131] For these studies, oolong tea extract was prepared according to the following procedure. To make tea extract from tea leaves, 2 g of tea leaves were extracted in deionized water at 100°C for 20 minutes with occasional mixing. The tea extract was filtered through a >10 μm cutoff filter to remove large particles. The extract was flash frozen in dry ice / ethanol and then lyophilized to obtain a dry, concentrated powder. The dried powder was weighed and resuspended in DMSO to make a concentrated 100 mg / ml stock solution. The stock solution was diluted into the aggregation reaction so that the DMSO concentration was less than 0.28% in the final reaction.

[0132] Oolong tea extract was prepared from oolong tea obtained from the Guan Yin Mountains (China) (MemorTea®; AuNutra Industries Inc., Chino, Calif.). PTI-00703® is a powdered extract made from an aqueous extract of the inner bark of Uncaria tomentosa obtained from the Amazon rainforest. Methods for preparing PTI-00703® are described, for example, in WO 98 / 51302. Blackcurrant extract containing 25% anthocyanins is a powdered extract prepared from European blackcurrant fruit (cassis fruit) and is obtained from Vitaquest (West Caldwell, NJ). Ethanol extracts of blackcurrant, oolong tea, and / or Uncaria tomentosa are within the scope of this disclosure.

[0133] Composition A containing oolong tea extract, Uncaria tomentosa extract, and blackcurrant extract (25% anthocyanins) was prepared as follows: Uncaria tomentosa extract (PTI-00703®) was first granulated using acacia gum as a binder to provide 206 mg of granulated Uncaria tomentosa extract (187.5 mg of cat's claw). An equal amount of oolong tea extract (187.5 mg) was then added, followed by 50 mg of blackcurrant extract (25% anthocyanins) to provide the test composition. Example 1: Disruption / Inhibition of Alzheimer's Disease Aβ Fibrils or Aggregates

[0134] The compositions described herein have been found to be potent disruptors / inhibitors of Aβ protein fibrils or aggregates. This example investigated the effectiveness of the compositions in causing disassembly / disruption / disaggregation of pre-formed amyloid fibrils of Alzheimer's disease (i.e., composed of Aβ1-42). Part A: Thioflavin T Fluorometry

[0135] In this study, the effect of the composition was determined using a thioflavin T fluorometer. Thioflavin T specifically binds to fibrillar amyloid, and this binding results in an increase in fluorescence at 485 nm that is directly proportional to the number of amyloid fibrils formed. The higher the fluorescence, the greater the number of amyloid fibrils formed (Nakai et al., Lab. Invest. 65:104-110, 1991; Levine III, Protein Sci. 2:404-410, 1993; Amyloid: Int. J. Exp. Clin. Invest. 2:1-6, 1995).

[0136] In this study, 40 μl of a 1 mg / ml solution (in distilled water) or prefibrillar human Aβ (rPeptide) was incubated for 3 days at 37°C, either alone (control) or in the presence of one of the compositions disclosed herein (Aβ:test composition weight ratios of 1:1, 1:0.1, 1:0.01, and 1:0.001). The final concentration of Aβ in the reaction was 0.4 mg / ml (88 μM) in phosphate-buffered saline (PBS), pH 7.4, and 0.02% sodium azide in a final volume of 100 μl. After 3 days of co-culture, 12.5 μl of each incubation mixture was transferred to a 96-well microtiter plate containing 37.5 μl of PBS and 200 μl of thioflavin T solution (i.e., 125 μM thioflavin T in 62.5 mM phosphate buffer, pH 6.8). Emitted fluorescence was read at 485 nm (444 nm excitation wavelength) using an ELISA plate fluorometer after subtraction with buffer alone or composition alone as blanks.

[0137] In a first series of studies, the following compositions were tested: [Table 1]

[0138] Figure 11 shows the results of a 3-day incubation with the following compositions: Composition 11.1; Composition 11.2; Composition 11.3; Composition 11.4, Composition 11.5, Composition 11.6, and Composition 11.7.

[0139] Incubation of Aβ1-42 with 11.5 (blackcurrant extract containing 25% anthocyanins) caused a dose-dependent disruption / disassembly of preformed Aβ1-42 fibrils. It was also observed that the disintegration rate was significantly greater with 11.5 compared to 11.4 (blackcurrant extract containing 15% anthocyanins).

[0140] In a second study, the following compositions were tested: [Table 2]

[0141] Figure 12 shows the results of a 3-day incubation with the following compositions: Composition 18.1; Composition 18.2; Composition 18.3; Composition 18.4, Composition 18.5, Composition 18.6, and Composition 18.7.

[0142] In a third study, the following compositions were tested: [Table 3]

[0143] Figure 13 shows the results of a 3 day incubation with the following compositions: Composition 19.1; Composition 19.2; Composition 19.3; Composition 19.5, Composition 19.6, Composition 19.7, Composition 19.8 and Composition 19.9.

[0144] In a fourth study, the following compositions were tested: [Table 4]

[0145] 14 shows the results of a 3-day incubation with the following compositions: Composition 21.1; Composition 21.2; Composition 21.3; Composition 21.5, Composition 21.6, and Composition 21.7. These results demonstrated that Composition A, a combination of blackcurrant extract, oolong tea extract, and cat's claw extract, is a potent disruptor / inhibitor of Alzheimer's disease-type Aβ fibrils and exerts its effects in a dose-dependent manner.

[0146] In another study involving the combination of oolong tea extract and cat's claw extract, 40 μl of a 1 mg / ml solution (in distilled water) or prefibrillar human Aβ (rPeptide) was incubated for 3 days at 37°C either alone (control) or in the presence of LOTE + PTI-00703® (test composition:Aβ weight ratios of 1:1 and 1:0.1) (referred to as "CognitiveClarity™"). The final concentration of Aβ in the reaction was 0.4 mg / ml (88 μM) in phosphate-buffered saline (PBS), pH 7.4 + 0.02% sodium azide, in a final volume of 100 μl. After 3 days of co-culture, 12.5 μl of each incubation mixture was transferred to a 96-well microtiter plate containing 37.5 μl of PBS and 200 μl of thioflavin T solution (i.e., 125 μM thioflavin T in 62.5 mM phosphate buffer, pH 6.8). Emitted fluorescence was read at 485 nm (444 nm excitation wavelength) using an ELISA plate fluorometer after subtraction with buffer alone or composition alone as blanks.

[0147] The results of a 3-day incubation of a combination of oolong tea extract and cat's claw extract are shown in Figure 1. Incubation of Aβ1-42 with "Cognitive Clarity" (i.e., the combination of LOTE and PTI-00703®) caused dose-dependent disruption / disassembly of preformed Aβ1-42 fibrils. At a test composition:Aβ weight ratio of 0.1:1, LOTE and PTI-00703 inhibited fibrils by 62.8%. At an equal weight equivalent (test composition:Aβ weight ratio of 1:1), there was a 93.6% inhibition of thioflavin T fluorescence. This study demonstrated that the combination of oolong tea extract and cat's claw extract is a potent disruptor / inhibitor of Alzheimer's disease-type Aβ fibrils, exerting its effects in a dose-dependent manner. Part B: Congo Red

[0148] The Congo Red binding assay quantifies the ability of a test composition to alter β-amyloid binding to Congo Red. In this assay, Aβ1-42 (prepared for the Thioflavin T assay) is incubated alone (control) or with increasing amounts of test composition for three days, then vacuum filtered through a 0.2 μm filter. The amount of Aβ1-42 retained on the filter is then quantified after staining the filter with Congo Red (125 μM Congo Red, 100 mM Tris, 50 mM NaCl, pH 7). After appropriate washing of the filter, any decrease in Congo Red color on the filter in the presence of the test composition (compared to Congo Red staining of amyloid protein in the absence of the test composition) indicates the ability of the test composition to reduce / alter the amount of aggregated Congo Red Aβ. Composition A, a combination of blackcurrant extract, oolong tea extract, and cat's claw extract, is expected to inhibit Aβ binding to Congo Red.

[0149] The ability of Aβ fibrils to bind Congo Red was determined for a combination of oolong tea extract and cat's claw extract in the absence or presence of increasing amounts of LOTE+PTI-00703® (test composition:Aβ weight ratios of 1:1 and 0.1:1). Results from a 3-day incubation are shown in Figure 1. LOTE+PTI-00703® caused a dose-dependent inhibition of Aβ binding to Congo Red. At a test composition:Aβ weight ratio of 0.1:1, LOTE+PTI-00703® inhibited Congo Red binding by 20.9% (p<0.001). At equal weight equivalents (test composition:Aβ weight ratio of 1:1), there was a 58.6% (p<0.001) inhibition of Congo Red binding.

[0150] Similar to the results of the Congo Red binding assay, this study also demonstrated that this combination of specific lead oolong tea extract (LOTE) and PTI-00703® was a potent disruptor / inhibitor of Aβ fibrils, as assessed by a thioflavin T fluorimetric assay, and exerted its effects in a dose-dependent manner (Figure 1). The combination of LOTE and PTI-00703® caused a dose-dependent reduction in thioflavin T binding (indicating Aβ fibril disruption / reduction) of 70% (p<0.001) at a test composition:Aβ weight ratio of 0.1:1 (Figure 1) and 95% at a test composition:Aβ weight ratio of 1:1 (Figure 1). Part C: Slide-based Congo Red conjugation, Thioflavin S, and electron microscopy

[0151] In the slide-based Congo Red assay, Congo Red dye is incubated with Aβ1-42, spotted onto a slide, and imaged under polarized light. Congo Red-bound amyloid fibrils emit a characteristic "apple-green birefringence" under polarized light. In this study, 0.4 mg / ml pre-fibrillated Aβ1-42 (as prepared for the Thioflavin T assay) is incubated for 3 days with or without the test composition. 10 μl of Congo Red solution (250 mg of Congo Red dye (54% purity; Sigma) dissolved in 1 L of dH2O) is added to 10 μl of the Aβ1-42- / + test composition and mixed by vortexing for 30 seconds. The sample is incubated with the Congo Red solution for 10 minutes at room temperature. The sample is then centrifuged at 2000 g for 3 minutes, and 10 μl of the supernatant is removed. 2 μl of glycerol is added to the pellet and mixed by pipetting up and down 15 times. After vortexing the sample, 10 μl of stained protein was spotted onto an 18-well 5 M M HTC® autoclavable blue slide. The sample was covered with a small circular coverslip and immediately imaged under polarized light. Images were captured with a Zeiss Axioscope 2 Plus microscope equipped with a Q-Imaging Retiga 1300 digital camera and an HBO 100 illuminator. It is expected that Composition A, a combination of blackcurrant extract, oolong tea extract, and cat's claw extract, will reduce and disaggregate / dissolve preformed fibrils of Aβ 1-42.

[0152] For the combination of oolong tea extract and cat's claw extract (Figure 2A (left panel)), a representative image shows that untreated Aβ1-42 has the characteristic apple-green birefringence and abundant fibrillar proteins uniformly distributed throughout the field of view. In Figure 2A (right panel), treatment with LOTE + PTI-00703 (0.1:1 weight ratio with Aβ42) resulted in significantly fewer Congo red-stained fibrils, indicating that this combination of plant extracts can reduce and disaggregate / dissolve preformed Aβ1-42 fibrils.

[0153] Similar to thioflavin T, thioflavin S is a related anionic dye that binds to fibrillar amyloid proteins and can be used to detect fibrillar proteins bound to glass microscope slides. In this study, pre-fibrillarized Aβ1-42 (as prepared for the thioflavin T assay) was incubated for 3 days with or without the test composition. 4 μl of 0.4 mg / ml Aβ1-42- / + test composition was spotted onto an 18-well 5 M M HTC® autoclavable blue slide. The sample was allowed to air dry for 2 hours. 10 μl of thioflavin S solution (31 mg of thioflavin S dissolved in 50 mL of dH2O) was gently applied to the dried protein on the slide. After staining the protein for 1 minute, the thioflavin solution was removed with a pipette. 40 μl of 70% ethanol solution was gently pipetted onto the stained protein for 1 minute to rinse. This solution was then gently removed with a pipette. 2 μl of Vectashield (Vector) mounting medium is applied to the stained protein, and then covered with a circular cover slip. Images are viewed under fluorescent light, and images are captured using a Zeiss Axioscope 2 Plus microscope equipped with an HBO100 illuminator and a Q-Imaging Retiga 1300 digital camera. It is expected that composition A, a combination of blackcurrant extract, oolong tea extract, and cat's claw extract, will significantly reduce the fluorescent fibrils of thioflavin S.

[0154] For the combination of oolong tea extract and cat's claw extract (Figure 2B (left panel)), a representative image of untreated Aβ1-42 shows abundant thioflavin S fluorescent fibrils evenly distributed throughout the field of view. Preformed Aβ1-42 fibrils incubated with LOTE+PTI-00703® (0.1:1 weight ratio with Aβ1-42) for 72 hours showed a substantial decrease in fluorescent fibrils (Figure 2B, right panel). In Figure 2B (right panel), a representative image shows that LOTE+PTI-00703 treatment resulted in the disappearance of thioflavin S-fluorescent fibrils.

[0155] Negative staining electron microscopy (EM) analysis is used to independently monitor the effectiveness of various compositions for disrupting preformed Aβ fibrils. In these experiments, preformed Aβ 1-42 fibrils (as prepared for the thioflavin T assay) are incubated in the absence (control) or presence of increasing concentrations of test compositions. After 3 days of incubation, 10 μl of the sample is spotted onto a grid, stained with 2% uranyl acetate, and visualized at 8,000x to 30,000x magnification using a JEOL1010 transmission electron microscope. EM analysis is expected to show that treatment with Composition A, a combination of blackcurrant extract, oolong tea extract, and cat's claw extract, will significantly reduce the number and dissolve clumped Aβ fibrils. These studies are expected to demonstrate that Composition A, a combination of blackcurrant extract, oolong tea extract, and cat's claw extract, is a potent Aβ disruptor / inhibitor.

[0156] For the combination of oolong tea extract and cat's claw extract (Figure 2C (bottom panel)), EM analysis confirmed the formation of Aβ fibrils without treatment (i.e., Figure 2C, control). Without treatment (2C, left panel), Aβ formed large aggregated fibrils that uniformly covered the field of view. These samples were also tested by thioflavin T assay, which confirmed thioflavin T fluorescence-positive fibrils. In the presence of LOTE + PTI-00703® (Figure 2C, right panel), the number of aggregated Aβ fibrils was significantly reduced and dissolved. These results correlate well with the thioflavin T fluorometry data, which showed a decrease in thioflavin T fluorescence with treatment, and the Congo red binding data, which showed a decrease in Congo red binding with treatment. Using these independent methodologies, we identified and validated Lead Oolong Tea Extract (LOTE) in combination with PTI-00703® (i.e., LOTE + PTI-00703®) as a potent Aβ disrupting / inhibitor. Example 2: Inhibition / Disruption of In Vitro Conversion of Aβ into β-Sheets Containing Fibrillar Structures Part A: Thioflavin T Fluorometry

[0157] To test whether the compositions described herein can inhibit Aβ β-sheet formation, the same Thioflavin T assay described in Example 1 was used, but instead Aβ1-40 was used as the substrate. Similar to Aβ1-42, Aβ1-40 forms Thioflavin T-positive aggregates, but requires incubation at 37°C for more than 24 hours with shaking for complete fibrillation. Because Aβ1-40 is in a non-fibrillar state at the start of the assay, this protein can aggregate in the presence of the composition to measure aggregation inhibition. Lyophilized human Aβ1-40 (rPeptide) was dissolved in dH2O to 1 mg / mL (220 μM). In separate test tubes, test composition stocks were prepared in PBS at various concentrations such that final reactions containing equal amounts of test composition stock and Aβ solution yielded test composition:Aβ weight ratios of 1:1, 0.5:1, 1:1, and 0.2:1, resulting in a final Aβ concentration of 0.5 mg / mL (110 μM). Reactions containing Aβ + test composition (or Aβ + PBS as a control for Aβ aggregation) are then incubated for 2 days. The incubation mixtures are diluted 1:10 to 0.05 mg / ml. 50 μL of Aβ and each diluted incubation mixture is transferred to a 96-well microtiter plate containing 200 μL of thioflavin T solution (i.e., 125 μM thioflavin T in 62.5 mM phosphate buffer, pH 6.8). Fluorescence is read at 485 nm (444 nm excitation wavelength) using an ELISA plate fluorometer after subtracting PBS buffer alone or composition alone as a blank. Composition A, a combination of blackcurrant extract, oolong tea extract, and cat's claw extract, is expected to inhibit Aβ β-sheet formation. Specifically, this study is expected to demonstrate that Composition A is a potent inhibitor of β-sheet-rich Aβ fibril formation, as assessed by thioflavin T fluorescence measurement, and may exert its effect in a dose-dependent manner.

[0158] LOTE+PTI-00703® was evaluated in a separate study in combination with oolong tea extract and cat's claw extract. The results of this study, presented in Figure 3, demonstrated that the present LOTE+PTI-00703® interferes with Aβ aggregation, as indicated by its ability to prevent fibril formation, as assessed by thioflavin T fluorescence measurements. At a test composition:Aβ weight ratio of 0.2:1, LOTE+PTI-00703® inhibited fibrils by 14.1%, and at 0.5:1, LOTE+PTI-00703® inhibited fibrils by 73% (p<0.001). At equal weight equivalents (test composition:Aβ weight ratio of 1:1), there was an 89.3% (p<0.001) inhibition of thioflavin T fluorescence. This study demonstrated that LOTE plus PTI-00703® is a potent inhibitor of β-sheet-rich Aβ fibril formation, as assessed by thioflavin T fluorescence measurement, and that this combination exerts its effect in a dose-dependent manner. Part B: Congo Red

[0159] To test whether the compositions described herein can inhibit Aβ β-sheet formation, the same Congo Red assay as described in Example 1 is utilized, but Aβ 1-40 is used instead as the substrate. In this assay, Aβ 1-40 (as prepared for the ThioT assay) and the test composition are incubated for 2 days and then vacuum filtered through a 0.2 μm filter. The amount of Aβ 1-40 retained on the filter is then quantified after staining the filter with Congo Red (125 μM Congo Red, 100 mM Tris, 50 mM NaCl, pH 7). After appropriate washing of the filter, any decrease in Congo Red color on the filter in the presence of the test composition (compared to Congo Red staining of amyloid protein in the absence of the test composition) indicates the ability of the test composition to reduce / alter the amount of aggregated Congo Red Aβ. Composition A, a combination of blackcurrant extract, oolong tea extract, and cat's claw extract, is expected to inhibit Aβ β-sheet formation. Specifically, it is expected that this study will demonstrate that Composition A is a potent inhibitor of Aβ fibrils, as assessed by Aβ fibril binding to Congo Red, and may exert its effects in a dose-dependent manner.

[0160] In a separate study, the ability of Aβ fibrils to bind to Congo red following the combination of oolong tea extract and cat's claw extract in the absence or presence of increasing amounts of LOTE+PTI-00703® (test composition:Aβ weight ratios of 1:1, 0.5:1, and 0.2:1) was determined. Results from a 2-day incubation are shown in Figure 3. LOTE+PTI-00703® caused a dose-dependent inhibition of Aβ binding to Congo red. At a test composition:Aβ weight ratio of 0.2:1, LOTE+PTI-00703 inhibited Congo red binding by 41.7%, and 0.5:1 LOTE+PTI-00703® inhibited Congo red binding by 62.5% (p<0.001). At equal weight equivalents (1:1 test composition:Aβ weight ratio), there was 83.3% (p<0.001) inhibition of Congo Red binding. Similar to the results of the Thioflavin T fluorimetric assay, this study also demonstrated that LOTE+PTI-00703® is a potent inhibitor of Aβ fibrils, as assessed by Aβ fibril binding to Congo Red, and exerts its effect in a dose-dependent manner. Part C: CD spectroscopy

[0161] CD spectroscopy was performed to determine the efficacy of the compositions described herein in inhibiting the formation of Aβ 1-40 β-sheet secondary structure under aggregation-prone conditions. Because β-sheet structure is characteristic of Aβ fibrils, monitoring protein secondary structure can provide further evidence of the composition's effectiveness in inhibiting aggregation. CD spectra of Aβ 1-40 samples with increasing concentrations of the composition were analyzed at 25°C using a JASCO Model J-810 spectropolarimeter. CD spectroscopy and thio-T assays were performed in parallel from the same sample preparations to correlate results from the two independent assays. CD spectral analysis is expected to demonstrate that Composition A, a combination of blackcurrant extract, oolong tea extract, and cat's claw extract, significantly inhibits the aberrant assembly of Aβ into fibrillar β-sheet assemblies and maintains Aβ 1-40 in a less pathogenic form.

[0162] CD spectra of Aβ1-40 treated with LOTE+PTI-00703® were performed for the combination of oolong tea extract and cat's claw extract. In Figure 4, the CD spectra of Aβ1-40 treated with LOTE+PTI-00703® showed dose-dependent inhibition of β-sheet, including fibrils. A minimum at 218 nm indicates the presence of β-sheet structure. A positive shift in ellipticity at 218 nm indicates less β-sheet structure. At a test composition:Aβ weight ratio of 0.2:1, LOTE+PTI-00703® showed 30.9% less β-sheet structure than untreated, and at 0.5:1, LOTE+PTI-00703® showed 53.9% less β-sheet structure than the control. At equal weight equivalents (1:1 test composition:Aβ weight ratio), there was 64.8% less β-sheet structure compared to the control. These data confirmed that LOTE+PTI-00703® has the significant ability to inhibit the aberrant assembly of Aβ into fibrillar β-sheet assemblies and maintain Aβ1-40 in a less pathogenic form. Example 3: Use of recombinant tau repeat domains for in vitro screening of tau aggregation inhibitors

[0163] In a previous study on in vitro screening to identify tau aggregation inhibitors, we found that under the same experimental conditions, the formation of paired helical filaments (PHFs) from commercially purchased full-length tau protein (Tau441; from rPeptide) was much slower (more than 11 days; data not shown) than that from the tau repeat domain (TauRD; containing Q244-E372 of Tau441) (more than 24 hours) (S. Barghorn et al., Methods Mol Biol, 299:35-51, 2005).

[0164] Due to its apparent short turnaround time and general aggregation properties, tau RD was used for in vitro screening of tau aggregation inhibitors. Because tau RD protein was not commercially available, tau RD was prepared according to the following procedure, also described in U.S. Patent Application Publication No. 2016 / 0250273, which is incorporated by reference for disclosure of this procedure. A cDNA fragment encoding human tau RD was cloned into a bacterial expression vector, and the construct was expressed in Escherichia coli. Bacterial clones showing high levels of tau RD expression were then selected for protein purification. The recombinant tau RD protein was then purified by thermostability treatment and cation exchange chromatography as described (S. Barghorn et al., Methods Mol Biol, 299:35-51, 2005) with minor modifications. Using this method, a protein yield of 20 mg per liter of bacterial culture was obtained. The aggregation and PHF formation of purified tau RDs were assessed and validated by independent assays, including thioflavin S (ThioS) fluorometry, a dye that fluoresces upon binding to fibrils, circular dichroism (CD) spectroscopy, a method for detecting changes in protein secondary structure, and electron microscopy. The results consistently show that tau RDs (10 μM) can form ThioS-positive, β-sheet-containing PHFs when incubated with equimolar heparin (Sigma-Aldrich, St. Louis, MO) at 37°C with shaking at 800–1000 rpm for more than a day.

[0165] In Figure 5A, tau RD (15 kDa) protein was purified from E. coli and assessed by SDS-PAGE / silver staining at a standard purity of >95%. CD spectroscopy was used to assess the non-aggregated and aggregated tau RD proteins. Figures 5B-5C show examples of CD spectroscopy of non-aggregated and aggregated tau RD proteins. Tau RD aggregates were prepared in the presence of an equimolar (10 μM) ratio of tau RD and heparin in 20 mM sodium phosphate buffer (pH 7.4) and incubated at 37 °C with shaking for 0-120 h. In the absence of heparin, the CD spectrum of non-aggregated tau RD was a random coil with minimal ellipticity at 195 nm (Figure 5B). In the presence of heparin, CD spectra showed a time-dependent conformational change in tau RD protein from random coil (minimum at 195 nm) at time 0 to β-sheet (minimum at 218 nm) between 20 and 120 hours (Figure 5C). Tau RD aggregation was monitored by time-dependent ThioS fluorescence measurements (Figure 5D). The results show the formation of ThioS-positive tau RD fibrils after 20 hours of incubation. In the absence of heparin without fibril formation, the tau RD ThioS signal was less than 200 arbitrary units (AU) of fluorescence at all time points. Tau fibril formation was confirmed by negative-stain EM (Figure 5E-G), which showed the formation of tau fibrils after 48 hours of incubation with heparin. Tau RD monomers at time 0 are shown in Figure 5E (bar = 200 nm). Tau RD fibril formation at 48 hours is shown in Figure 5F-G (bar = 50 nm). Both straight filaments and paired helical filaments were found. Example 4: Identification of novel tau aggregation inhibitors by thioflavin S fluorimetric screening

[0166] A well-known method for measuring fibril formation is the thioflavin T (ThioT) fluorometry (H. Naiki et al., Lab. Invest. 65:104-110, 1991; H. Levine III, Protein Sci. 2:404-410, 1993; H. Levine III, Amyloid 2:1-6, 1995; H. Naiki and K. Nakakuki, Lab. Invest. 74:374-383, 1996). ThioT is known to bind to fibrillar proteins, and an increase in fluorescence correlates with increased fibril formation, while a decrease in fluorescence correlates with a decrease in fibrils due to disassembly and / or disruption. The assay was modified by replacing ThioT with Thioflavin S (ThioS), a related anionic dye with similar properties. The latter has been shown to be sensitive and reproducible for the quantification of tau PHFs (data not shown; P. Friedhoff et al., Biochemistry, 37(28):10223-30, 1998). ThioS fluorescence measurements are used to assess whether the above compositions can cause fibril disassembly / disruption.

[0167] Aggregated tau fibrils are prepared in the presence of an equimolar ratio of tau RD and heparin (10 μM each) in 20 mM sodium phosphate buffer (pH 7.4). The reaction mixture is incubated at 37°C with shaking (800-1000 rpm) for 24-72 hours. Test compositions are tested at various weight-to-weight concentrations with 0.14 mg / ml tau RD containing heparin. Identical reaction mixtures (plus increasing concentrations of test composition) are set up in parallel without tau RD to serve as background controls.

[0168] After 24-72 hours of co-incubation, 50 μl of each culture mixture was transferred to a black 96-well microtiter plate (Santa Cruz Biotechnology, Inc., Dallas, Tex.) containing 50 μl of phosphate-buffered saline (PBS; Sigma-Aldrich, St. Louis, Mo.) and 25 μl of thioflavin S solution (500 mM thioflavin S in PBS, pH 7.4; Sigma-Aldrich, St. Louis, Mo.). Fluorescence was read at 485 nm (444 nm excitation wavelength) using an ELISA plate fluorometer after subtracting buffer-only or composition-only blanks.

[0169] IC for 50% inhibition of tau aggregation 50 is calculated by nonlinear regression [(log [inhibitor] vs. normalized response; variable slope)] using Prism version 5 software (GraphPad Software). It is expected that Composition A, a combination of blackcurrant extract, oolong tea extract, and cat's claw extract, will significantly reduce Thio S-positive tau aggregates. It is also expected that this study will demonstrate that Composition A is an inhibitor of tau aggregation / fibril formation that can be used as a novel therapeutic agent for tauopathies.

[0170] In another study, over 25 tea extracts were tested for inhibition of ThioS-positive tau aggregates. The majority of teas were oolong tea extracts. Figure 6 shows examples of ThioS fluorescence of aggregated tau in the presence of several oolong tea extracts, chamomile tea, and green tea. Increasing concentrations of extracts were incubated with 0.14 mg / ml (10 μM) tau RD and equimolar heparin for 24–72 hours with shaking at 1000 rpm and 37°C. Lead Oolong Tea Extract (LOTE) inhibited tau aggregation more than other oolong, chamomile, or green tea extracts. LOTE inhibited tau aggregation by 50% (IC) at a 0.23:1 (weight:weight) ratio of its composition to tau. 50 This experiment demonstrates that LOTE has a superior ability to inhibit the abnormal assembly of tau into filamentous pathogenic aggregates compared to other compositions.

[0171] After identifying tea extracts with superior tau aggregation inhibition, Lead Oolong Tea Extract (LOTE) was incubated alone or in combination with PTI-00703® in a tau aggregation inhibition assay. The combination of specific oolong tea with PTI-00703® inhibited ThioS-positive tau aggregates better than LOTE or PTI-00703® alone ( * Significant by paired t-test, p<0.05) (Figure 7). This combination of Reed Oolong tea extract and PTI-00703® is a potent inhibitor of tau aggregation / fibrillogenesis, which can be used as a novel treatment for tauopathies. Example 5: Confirmation of tau aggregation inhibition by analysis of protein secondary structure by circular dichroism (CD) spectroscopy

[0172] CD is a powerful dye-independent method for identifying changes in protein structure. CD measures the difference in absorption between left-handed and right-handed circularly polarized light. Proteins contain asymmetric elements, resulting in a distinct CD signal measured in units of ellipticity. Thioflavin S fluorescence indicates that tau aggregates have an ellipticity minimum at 218 nm, characteristic of β-sheet-containing proteins. Non-aggregated tau has an ellipticity minimum at 195 nm, indicative of a random coil structure. CD spectroscopy is performed to determine the efficacy of any one of the compositions described herein in inhibiting the formation of β-sheet secondary structure in tau RD under aggregation-prone conditions. CD spectra are obtained from samples containing + / - tau RD and increasing concentrations of the compound and analyzed at 25°C on a JASCO Model J-810 spectropolarimeter. CD spectroscopy and the Thioflavin S assay are run in parallel from the same sample preparation to correlate the results of the two independent assays. Composition A, a combination of blackcurrant extract, oolong tea extract, and cat's claw extract, is predicted to inhibit the abnormal assembly of tau into fibrillar beta-sheet assemblies and maintain tau in a non-pathogenic soluble random coil form.

[0173] In another study, a combination of oolong tea extract and cat's claw extract was evaluated. In Figure 8, tau was treated with lead oolong tea extract (LOTE) or chamomile tea extract for 48 hours. LOTE-treated tau showed a dose-dependent inhibition of conversion to β-sheet-containing fibrils. At the highest concentration of LOTE treatment, tau remained in a soluble random-coil form with a minimum size of approximately 195 nm. Conversely, at the highest concentration of chamomile tea extract treatment, tau still transformed into an aggregated, β-sheet structure, similar to untreated controls. These data confirm that LOTE, more than other compositions, has the significant ability to inhibit the abnormal assembly of tau into fibrillar β-sheet assemblies and maintain tau in a non-pathogenic, soluble random-coil form. Example 6: Inhibition of tau protein fibril formation and disaggregation of preformed tau fibrils as determined by negative staining electron microscopy (EM)

[0174] EM analysis is used to independently monitor the effectiveness of any one of the compositions described herein for inhibiting tau fibril formation. In these experiments, tau fibrils are assembled by incubating an equimolar ratio of tau RD protein and heparin (10 μM each) in the absence (control) or presence of increasing concentrations of the test composition. After two days of incubation, samples are spotted onto grids, stained with 2% uranyl acetate, and visualized at 8,000x to 30,000x magnification using a JEOL1010 transmission electron microscope. Composition A, a combination of blackcurrant extract, oolong tea extract, and cat's claw extract, is expected to inhibit tau fibril formation by EM analysis. The results of Composition A correlate well with ThioS fluorescence measurements and CD analysis, which are expected to show reduced ThioS fluorescence and reduced β-sheet structure. EM analysis is also expected to demonstrate that Composition A inhibits tau aggregation and also disaggregates preformed tau fibrils. It is expected that these studies will demonstrate that Composition A is a potent inhibitor of tau aggregation / fibril formation.

[0175] In another study, EM analysis of a combination of oolong tea extract and cat's claw extract (Figure 9, left panel) confirmed the formation of tau fibrils in the absence of treatment. Without treatment, tau formed a mixture of paired straight and helical filaments similar to those seen in human tauopathies (V.M. Lee et al., Ann. Rev. Neurosci. 24:1121-159, 2001). These samples were also tested by ThioS assay and CD, which confirmed ThioS fluorescence positivity and β-sheet structure. In the presence of PTI-00703® and PTI-00703® + LOTE (Figure 9, center and right panels), tau fibrils became shorter and sparser, indicating inhibition of tau fibril formation. Fewer fibrils were evident in tau samples treated with both PTI-00703® and LOTE. These results correlated well with ThioS fluorescence measurements and CD analysis, which showed decreased ThioS fluorescence and reduced β-sheet structure. Using three independent methodologies, we identified and validated PTI-00703 + Lead Oolong Tea Extract as a potent inhibitor of tau aggregation / fibril formation.

[0176] Using EM analysis, preformed tau fibrils were shown to rapidly disaggregate in the presence of both PTI-00703® and LOTE. Tau RDs were incubated with equimolar heparin to form fibrils as described in the previous assay. Tau RDs were diluted with or without the test composition and incubated with shaking at 37°C for various time points. At each time point, tau RDs + / - test compounds were assayed for ThioS fluorescence and flash-frozen for EM analysis. In Figure 10 (left panel), without treatment, prefibrillar tau remained long filaments. In the next panel, prefibrillar tau was found to rapidly disaggregate in the presence of PTI-00703® + LOTE. After as little as 15 minutes of incubation with PTI-00703® + LOTE, tau fibrils were shorter and sparser than those without treatment. Disruption of tau fibrils was also confirmed by ThioS assay (data not shown). These data indicate that the PTI-00703®+LOTE composition not only inhibits tau aggregation but is also able to disaggregate pre-formed tau fibrils. Example 7: Dissolution of Cat's Claw / Oolong Tea Extract with the Addition of Blackcurrant Extract

[0177] In the following example, the solubility of cat's claw / oolong tea extract and cat's claw / oolong tea extract with added blackcurrant extract was compared. A visual analysis was performed using a glass of water to observe the dissolution of cat's claw / oolong tea and cat's claw / oolong tea with added 50 mg of European blackcurrant. Surprisingly, the addition of blackcurrant made the cat's claw / oolong tea extract much more water-soluble. It also made the composition of cat's claw / oolong tea extract with added blackcurrant taste much better (sweeter, less bitter, and less astringent).

[0178] Figure 15 shows a capsule of cat's claw / oolong tea extract in front of a glass of water (left side of the figure), and on the right is a capsule of the same cat's claw / oolong tea extract plus 50 mg of European blackcurrant extract.

[0179] Figure 16 shows one capsule in front of a glass of water. The left side contains cat's claw / oolong tea extract, and the right side contains a combination of cat's claw / oolong tea extract / blackcurrant.

[0180] Figure 17A shows the agglomeration (within 5 seconds) and water insolubility of the cat's claw / oolong tea extract combination. Figure 17B shows the agglomeration (within 5 seconds) of the cat's claw / oolong tea / black currant combination. Figure 17C shows that within 10 seconds, the cat's claw / oolong tea / black currant combination surprisingly begins to self-dissolve in water.

[0181] Figure 18A shows that after stirring with a spoon, the cat's claw / oolong tea extract (left) remains insoluble with clumps of insoluble material at the top of the glass of water. Meanwhile, the addition of blackcurrant (right), which turns the water a deep purple, also makes the mixture much more water-soluble, with little to no clumps of material at the top of the glass. Figure 18B is a close-up of the cat's claw / oolong tea extract, showing that after stirring, there are large clumps of insoluble material at the top of the glass, making it insoluble in water. Figure 18C is a close-up of the cat's claw / oolong tea / blackcurrant combination, showing that it is mostly dissolved in water after stirring. The addition of blackcurrant also makes the solution taste much sweeter (less bitter and less astringent). Example 8: In vivo studies of cognition and memory improvement

[0182] Further in vivo studies will be used to test any one of the compositions described herein for its effectiveness in reducing brain "plaque and tangle" load and improving cognition and memory. 40 to 60 men and women will be selected for clinical trials. The subjects will suffer from age-associated memory impairment (AAMI) and expect their memory loss symptoms to worsen within the 6-month study period, but will otherwise be in good health.

[0183] This study includes a placebo group; that is, subjects are divided into two groups, one of which receives a test composition, any one of the compositions described herein formulated as a capsule (e.g., two 375 mg or 670 mg capsules containing Composition A with a meal, preferably lunch), and the other a placebo (two capsules containing capsules without the active ingredient of the study product). Subjects are benchmarked for memory, cognition, concentration, attention, reasoning, and other symptoms associated with mild cognitive impairment (MCI). Subjects in the test group receive therapeutic doses of the combined test product extract or placebo for six months, and analyses of short-term memory, cognition, concentration, and attention are performed at 0, 1, 3, and 6 months of treatment. Accurate records of memory, attention, and attention are kept for both groups regarding benchmark symptoms, and these results are compared at the end of the study. Results are also compared between members of each group. Furthermore, each subject's results are compared to the symptoms reported by each subject before the study began. The activity of the test compositions used is demonstrated by attenuation of typical cognitive decline, decline in short-term memory, cognition, concentration and attention, and / or related behavioral impairments associated with age-associated memory impairment (AAMI). Composition A, a combination of blackcurrant extract, oolong tea extract, and cat's claw extract, is expected to reduce the brain's "plaque and tangle" burden and / or improve cognition and memory.

[0184] The present disclosure should not be limited in terms of the specific embodiments described in this application, which are intended as single illustrations of individual aspects of the present disclosure. Not all various embodiments of the present disclosure are described herein. As will be apparent to those skilled in the art, many modifications and variations of the present disclosure can be made without departing from its spirit and scope. Functionally equivalent methods and apparatuses within the scope of the present disclosure, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing description. Such modifications and variations are intended to fall within the scope of the appended claims. The present disclosure should be limited only by the terms of the appended claims, along with the full scope of equivalents to which such claims are entitled.

[0185] It is to be understood that this disclosure is not limited to particular uses, methods, reagents, compounds, compositions or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0186] Furthermore, when features or aspects of the present disclosure are described in terms of a Markush group, those skilled in the art will recognize that the present disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0187] As will be understood by those skilled in the art, for any and all purposes, particularly with respect to providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations thereof. A recited range can be easily recognized as fully descriptive and validating at least equal halves, thirds, quarters, fifths, tenths, etc. of the same range. As a non-limiting example, each range discussed herein can be readily broken down into lower, middle, and upper thirds, etc. As will be understood by those skilled in the art, all terms such as "up to," "at least," "greater than," "less than," etc., include the recited numbers and refer to ranges that can be subsequently broken down into subranges as described above. Finally, as will be understood by those skilled in the art, ranges include individual members. Thus, for example, a group having 1 to 3 members refers to groups having 1, 2, or 3 members. Similarly, a group having 1 to 5 members refers to groups having 1, 2, 3, 4, or 5 members.

Claims

[Claim 1] 1. A composition comprising a blackcurrant extract, an Uncaria tomentosa extract, and an oolong tea extract, the blackcurrant extract comprises from about 10% w / w to about 35% w / w of proanthocyanidins, anthocyanidins, anthocyanins, or combinations thereof; the Uncaria tomentosa extract is PTI-00703, The oolong tea extract is derived from a water extract of oolong tea leaves, The composition contains Uncaria tomentosa extract and oolong tea extract and has higher solubility in water than a composition not containing blackcurrant extract. composition.

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