US 20010047032A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2001/0047032 A1 Castillo et al. (43) Pub. Date: Nov. 29, 2001

(54) POLYHYDROXYLATED AROMATIC Related U.S. Application Data COMPOUNDS FOR THE TREATMENT OF AMYLOIDOSIS AND ALPHA-SYNUCLEIN (63) Non-provisional of provisional application No. FBRIL DISEASES 60/173,958, filed on Dec. 30, 1999. (76) Inventors: Gerardo M. Castillo, Seattle, WA Publication Classification (US); Paula Y. Choi, Bothell, WA (US); Alan D. Snow, Lynnwood, WA (51) Int. Cl." ...... A61K 3177042; A61K 31/70; (US) A61K 31/66 (52) U.S. Cl...... 514/453; 514/456; 514/23; Correspondence Address: 514/643; 514/734; 514/712; HELLER EHRMAN WHITE & MCAULIFFE 514/27; 514/129 LLP 275 MIDDLEFIELD ROAD (57) ABSTRACT MENLO PARK, CA 94025-3506 (US) Polyhydroxylated aromatic compounds, and compositions containing them, are useful for the treatment of amyloidosis, (21) Appl. No.: 09/748,748 especially Alzheimer's disease, and for the treatment of diseases characterized by C-Synuclein fibril formation, espe (22) Filed: Dec. 26, 2000 cially Lewy body disease and Parkinson's disease. US 2001/0047032A1 Nov. 29, 2001

POLYHYDROXYLATED AROMATIC COMPOUNDS circulating precursor protein may result from overproduc FOR THE TREATMENT OF AMYLOIDOSIS AND tion of either intact or aberrant molecules (for example, in ALPHA-SYNUCLEIN FIBRIL DISEASES plasma cell dyscrasias), reduced degradation or excretion (serum amyloid A in Some Secondary amyloid syndromes BACKGROUND OF THE INVENTION and beta-microglobulin in long-term hemodialysis), or genetic abnormalities associated with variant proteins (for 0001) 1. Field of the Invention example, familial amyloidotic polyneuropathy). Proteolysis 0002 This invention relates to the use of certain poly of a larger protein precursor molecule occurs in many types hydroxylated aromatic compounds, and compositions con of amyloidosis, resulting in the production of lower molecu taining them, for the treatment of amyloidosis, especially lar weight fragments that polymerize and assume a beta Alzheimer's disease, and the treatment of diseases charac pleated sheet conformation as tissue deposits, usually in an terized by C-Synuclein fibril formation, especially Lewy extracellular location. The precise mechanisms involved and body disease and Parkinson's disease. the aberrant causes leading to changes in proteolytic pro cessing and/or translational modification are not known in 0003 2. Description of the Related Art most amyloids. 0004 Amyloid and Amyloidosis 0008 Systemic amyloids which include the amyloid 0005 Amyloid is a generic term referring to a group of asSociated with chronic inflammation, various forms of diverse but specific extracellular protein deposits which all malignancy and familial Mediterranean fever (i.e. AA amy have common morphological properties, Staining character loid or inflammation-associated amyloidosis) (Benson and istics, and X-ray diffraction spectra. Regardless of the nature Cohen, Arth. Rheum. 22:36-42, 1979; Kamei et al., Acta of the amyloid protein deposited all amyloids have the Path. Jpn. 32:123-133, 1982; McAdam et al., Lancet 2:572 following characteristics: 1) showing an amorphous appear 573, 1975; Metaxas, Kidney Int. 20:676-685, 1981), and the ance at the light microscopic level, appearing eosinophilic amyloid associated with multiple myeloma and other B-cell using hematoxylin and eosin stains; 2) staining with Congo dyscrasias (i.e. AL amyloid) (Harada et al., J. Histochem. red and demonstrating a red/green birefringence as viewed Cytochem. 19:1-15, 1971), as examples, are known to under polarized light (Puchtler et al., J. Histochem. involve amyloid deposition in a variety of different organs Cytochem. 10:355-364, 1962), 3) containing a predominant and tissues generally lying outside the central nervous beta-pleated sheet Secondary structure, and 4) ultrastructur System. Amyloid deposition in these diseases may occur, for ally consisting of non-branching fibrils of indefinite length example, in liver, heart, Spleen, gastrointestinal tract, kidney, and with a diameter of 7-10 nm. skin, and/or lungs (Johnson et al., N. Engl. J. Med. 321:513 518, 1989). For most of these amyloidoses, there is no 0006 Amyloidoses today are classified according to the apparent cure or effective treatment and the consequences of Specific amyloid protein deposited. The amyloids include, amyloid deposition can be detrimental to the patient. For but are not limited to, the amyloid associated with Alzhe example, amyloid deposition in the kidney may lead to renal imer's disease, Down's Syndrome and hereditary cerebral failure, whereas amyloid deposition in the heart may lead to hemorrhage with amyloidosis of the Dutch type (where the heart failure. For these patients, amyloid accumulation in Specific amyloid is referred to as beta-amyloid protein or Systemic organs leads to eventual death generally within 3-5 Af), the amyloid associated with chronic inflammation, years. Other amyloidoses may affect a single organ or tissue various forms of malignancy and familial Mediterranean such as observed with the AP amyloid deposits found in the fever (where the specific amyloid is referred to as AA brains of patients with Alzheimer's disease and Down's amyloid or inflammation-associated amyloid), the amyloid syndrome: the PrP amyloid deposits found in the brains of asSociated with multiple myeloma and other B-cell dyscra patients with Creutzfeldt-Jakob disease, Gerstmann-Straus sias (where the specific amyloid is referred to as AL amy sler Syndrome, and kuru; the islet amyloid (amylin) deposits loid), the amyloid associated with type II diabetes (where the found in the islets of Langerhans in the pancreas of 90% of Specific amyloid is referred to as amylin or islet amyloid), patients with type II diabetes (Johnson et al., N. Engl. J. Med. the amyloid associated with the prion diseases including 321:513-518, 1989; Lab. Invest. 66:522 535, 1992); the Creutzfeldt-Jakob disease, Gerstmann-Straussler Syndrome, beta-microglobulin amyloid deposits in the medial nerve kuru, and Scrapie (where the specific amnyloid is referred to leading to carpal tunnel Syndrome as observed in patients as PrP amyloid), the amyloid associated with long-term undergoing long-term hemodialysis (Geyjo et al, Biochem. hemodialysis and carpal tunnel Syndrome (where the spe Biophys. Res. Comm. 129:701-706, 1985; Kidney Int. cific amyloid is referred to as beta-microglobulin amyloid), 30:385-390, 1986); the-prealbumin/transthyretin amyloid the amyloid associated with Senile cardiac amyloid and observed in the hearts of patients with Senile cardiac amy familial amyloidotic polyneuropathy (where the specific loid; and the prealbumin/transthyretin amyloid observed in amyloid is referred to as prealbumin or transthyretin amy peripheral nerves of patients who have familial amyloidotic loid), and the amyloid associated with endocrine tumors polyneuropathy (Skinner and Cohen, Biochem. Biophys. Such as medullary carcinoma of the thyroid (where the Res. Comm. 99:1326-1332, 1981; Saraiva et al., J. Lab. Clin. Specific amyloid is referred to as variants of procalcitonin). Med. 102:590-603, 1983; J. Clin. Invest. 74: 104-119, 1984; 0007 Although amyloid deposits in clinical conditions Tawara et al., J. Lab. Clin. Med. 98:811-822, 1989). share common physical properties relating to the presence of a beta-pleated sheet conformation, it is now clear that many 0009 Alzheimer's Disease and the Aging Population different chemical types exist and additional ones are likely 0010 Alzheimer's disease is a leading cause of dementia to be described in the future. It is currently thought that there in the elderly, affecting 5-10% of the population over the age are Several common pathogenetic mechanisms that may be of 65 years (A Guide to Understanding Alzheimer's Disease operating in amyloidosis in general. In many cases, a and Related Disorders, Jorm, ed., New York University US 2001/0047032A1 Nov. 29, 2001

Press, New York, 1987). In Alzheimer's disease, the parts of patients with Alzheimer's disease. In addition, Alzheimer's the brain essential for cognitive processes Such as memory, disease is characterized by the presence of numerous neu attention, language, and reasoning degenerate, robbing Vic rofibrillary "tangles', consisting of paired helical filaments tims of much that makes us human, including independence. which abnormally accumulate in the neuronal cytoplasm In Some inherited forms of Alzheimer's disease, onset is in (Grundke-Iqbal et al., Proc. Natl. Acad. Sci. USA 83:4913 middle age, but more commonly, Symptoms appear from the 4917, 1986; Kosik et al., Proc. Natl. Acad. Sci. USA mid-60's onward. Alzheimer's disease today affects 4-5 83:4044-4048, 1986; Lee et al., Science 251:675-678, 1991). million Americans, with slightly more than half of these The pathological hallmark of Alzheimer's disease is there people receiving care at home, while the others are in many fore the presence of "plaques' and "tangles', with amyloid different health care institutions. The prevalence of Aizhe imer's disease and other dementias doubles every 5 years being deposited in the central core of the plaques. The other beyond the age of 65, and recent Studies indicate that nearly major type of lesion found in the Alzheimer's disease brain 50% of all people age 85 and older have symptoms of is the accumulation of amyloid in the walls of blood vessels, Alzheimer's disease (1999 Progress Report On Alzheimer's both within the brain parenchyma and in the walls of Disease, National Institute on Aging/National Institute of meningeal vessels which lie outside the brain. The amyloid Health). 13% (33 million people) of the total population of deposits localized to the walls of blood vessels are referred the United States are age 65 and older, and this percentage to as cerebrovascular amyloid or congophilic angiopathy will climb to 20% by the year 2025 (1999 Progress Report (Mandybur, J. Neuropath. Exp. Neurol. 45:79-90, 1986; On Alzheimer's Disease). Pardridge et al., J. Neurochem. 49:1394-1401, 1987). 0.011 Alzheimer's disease also puts a heavy economic 0016 For many years there has been an ongoing scien burden on Society. A recent Study estimated that the cost of tific debate as to the importance of “amyloid” in Alzheimer's caring for one Alzheimer's disease patient with Severe disease, and whether the “placques' and "tangles' character cognitive impairments at home or in a nursing home, is more istic of this disease were a cause or merely a consequence of than $47,000 per year (A Guide to Understanding Alzhe the disease. Within the last few years, studies now indicate imer's Disease and Related Disorders). For a disease that that amyloid is indeed a causative factor for Alzheimer's can Span from 2 to 20 years, the Overall cost of Alzheimer's disease and should not be regarded as merely an innocent disease to families and to Society is staggering. The annual bystander. The Alzheimer's AB protein in cell culture has economic toll of Alzheimer's disease in the United States in been shown to cause degeneration of nerve cells within Short terms of health care expenses and lost wages of both patients periods of time (Pike et al., Br. Res. 563:311-314, 1991; J. and their caregivers is estimated at S80 to S100 billion (1999 Neurochem. 64:253-265, 1995). Studies suggest that it is the Progress Report On Alzheimer's Disease). fibrillar structure (consisting of a predominant beta-pleated 0012 Tacrine hydrochloride (“Cognex”), the first FDA sheet Secondary structure), characteristic of all amyloids, approved drug for Alzheimer's disease, is a acetylcholinest that is responsible for the neurotoxic effects. A? has also erase inhibitor (Cutler and Sramek, N. Engl. J Med. 328:808 been found to be neurotoxic in Slice cultures of hippocampus 810, 1993). However, this drug has showed limited success (Harrigan et al., Neurobliol. Aging 16:779-789, 1995) and in producing cognitive improvement in Alzheimer's disease induces nerve cell death in transgenic mice (Games et al., patients and initially had major side effects Such as liver Nature 373:523-527, 1995; Hsiao et al., Science 274:99-102, toxicity. The Second more recently FDA approved drug, 1996). Injection of the Alzheimer's AB into rat brain also donepezil (“Aricept”), which is also an acetylcholinesterase causes memory impairment and neuronal dysfunction inhibitor, is more effective tham tacrine, by demonstrating (Flood et al., Proc. Natl. Acad. Sci. USA 88:3363-3366, Slight cognitive improvement in Alzheimer's disease 1991; Br. Res. 663:271-276, 1994). patients (Barner and (Gray, Ann. Pharmacotherapy 32:70 0017 Probably, the most convincing evidence that AB 77, 1998; Rogers and Friedhoff, Eur: Neuropsych. 8:67-75, amyloid is directly involved in the pathogenesis of Alzhe 1998), but is not believed to be a cure. Therefore, it is clear imer's disease comes from genetic Studies. It has been that there is a need for more effective treatments for Alzhe discovered that the production of AB can result from muta imer's disease patients. tions in the gene encoding, its precursor, beta amyloid 0013 Amyloid as a Therapeutic Target for Alzheimer's precursor protein (Van Broeckhoven et al., Science Disease 248:1120-1122, 1990; Murrell et al., Science 254:97-99, 0.014) Alzheimer's disease is characterized by the depo 1991; Haass et al., Nature Med. 1: 1291-1296, 1995). The Sition and accumulation of a 39-43 amino acid peptide identification of mutations in the beta-amyloid precursor termed the beta-amyloid protein, A? or B/A4 (Glenner and protein gene which causes early onset familial Alzheimer's Wong, Biochem. Biophys. Res. Comm. 120:885-890, 1984; disease is the Strongest argument that amyloid is central to Masters et al., Proc. Natl. Acad. Sci. USA 82:4245-4249, the pathogenetic proceSS underlying this disease. Four 1985; Husby et al., Bull. WHO 71:105-108, 1993). A? is reported disease-causing mutations have now been discov derived by protease cleavage from larger precursor proteins ered which demonstrate the importance of AB in causing termed beta-amyloid precursor proteins (or BPPs) of which familial Alzheimer's disease (reviewed in Hardy, Nature there are Several alternatively spliced variants. The most Genet. 1:233-234, 1992). All of these studies suggest that abundant forms of the BPPs include proteins consisting of providing a drug to reduce, eliminate or prevent fibrillar AB 695, 751 and 770 amino acids (Tanzi et al., Nature 331:528 formation, deposition, accumulation ancdor persistence in 530, 1988; Kitaguchi et al., Nature 331:530-532, 1988; the brains of human patients will Serve as an effective Ponte et al., Nature 331:525-527, 1988). therapeutic. 0.015 The small AB peptide is a major component which 0018 Discovery and identification of new compounds or makes up the amyloid deposits of "plaques' in the brains of agents as potential therapeutic agents to arrest amyloid US 2001/0047032A1 Nov. 29, 2001

deposition, accumulation and/or persistence that occurs in Selected from the group consisting of the compounds of Alzheimer's disease and other amyloidoses are desperately formula A, formula B, formula C, formula D, and formula E: Sought.

0019 Parkinson's Disease and C-Synuclein Fibril For Formula A mation OH 0020 Parkinson's disease is a neurodegenerative disor R2 OH der that is pathologically characterized by the presence of intracytoplasmic Lewy bodies (Lewy in Handbuch der Neurologie, M. Lewandowski, ed., Springer, Berlin, pp. X OH 920-933, 1912; Pollanen et al., J. Neuropath. Exp. Neurol. 52:183-191, 1993), the major components of which are R1 filaments consisting of C-Synuclein (Spillantini et al., Proc. Formula B Natl. Acad. Sci. USA 95:6469-6473, 1998; Arai et al., OH Neurosc. Lett. 259:83-86, 1999), an 140-amino acid protein (Ueda et al., Proc. Natl. Acad. Sci. USA 90:11282-11286, R2 OH 1993). Two dominant mutations in C-Synuclein causing familial early onset Parkinson's disease have been described Suggesting that Lewy bodies contribute mechanistically to R OH the degeneration of neurons in Parkinson's disease OPoly X meropoulos et al., Science 276:2045-2047, 1997; Kruger et Formula C al., Nature Genet. 18: 106-108, 1998). Recently, in vitro R2 Studies have demonstrated that recombinant C.-Synuclein can indeed form Lewy body-like fibrils (Conway et al., Nature Y. O X Med. 4:1318-1320, 1998; Hashimoto et al., Brain Res. 799:301-306, 1998; Nahri et al., J. Biol. Chem. 274:9843 9846, 1999). Most importantly, both Parkinson's disease Ri OH linked C-Synuclein mutations accelerate this aggregation proceSS which Suggests that Such in Vitro Studies may have OH O relevance for Parkinson's disease pathogenesis. C-Synuclein Formula D OH aggregation and fibril formation fulfills of the criteria of a nucleation-dependent polymerization process (Wood et al., J. Biol. Chem. 274: 19509-19512, 1999). In this regard C-synuclein fibril formation resembles that of Alzheimer's beta-amyloid protein (AB) fibrils. C-Synuclein recombinant protein, and non-amyloid component (known as NAC-P), which is a 35-amino acid peptide fragment of C-Synuclein, both have the ability to form fibrils when incubated at 37 C., and are positive with amyloid Stains Such as Congo red (demonstrating a red/green birefringence when viewed Formula E under polarized light) and Thioflavin S (demonstrating posi OH tive fluorescence) (Hashimoto et al., Brain Res. 799:301 306, 1998: Ueda et al., Proc. Natl. Acad. Sci. USA 90:11282 OH 11286, 1993). HO O 0021 Parkinson's disease C-synuclein fibrils, like the AB R fibrils of Alzheimer's disease, also consist of a predominant beta-pleated sheet structure. We believe, therefore, that compounds found to inhibit Alzheimer's disease AB amyloid OR fibril formation can also be anticipated to be effective in the OH inhibition of C-synuclein fibril formation. These compounds would therefore also serve as therapeutics for Parkinson's disease, in addition to having efficacy as a therapeutic for 0024 where: Alzheimer's disease and other amyloid disorders. 0025 R is selected from the group consisting of 0022. The disclosures of these and other documents hydrogen, 2,3-dihydroxybenzoyl, 3,4-dihydroxy referred to throughout this application are incorporated benzoyl, 2,34-trihydroxybenzoyl, and 3,4,5-trihy herein by reference. droxybenzoyl;

SUMMARY OF THE INVENTION 0026 R' is hydrogen or OH: 0027 R and R are independently selected from 0023. In a first aspect, this invention provides a method hydrogen and non-interfering Substituents, of treating amyloidosis in a mammal Suffering therefrom, comprising administration to the mammal of a therapeuti 0028 X is selected from hydrogen and the group cally effective amount of an isolated pure compound consisting of US 2001/0047032A1 Nov. 29, 2001

0029) (a) hydroxy, amino, C, alkylamino, di(C, 0037) and the pharmaceutically acceptable Salts thereof. alkyl)amino, and cycloamino, 0038. In preferred embodiments of this first aspect, only 0030 (b) C-2 alkyl, C-2 alkoxy, C-2 alky lthio, and C-2 alkylcarboxyl, each optionally one Such compound is administered; the mammal is a Substituted with 1 to 5 moieties selected from the human; and the amyloidosis is Selected from the group group consisting of halogen, hydroxy, mercapto, consisting of Alzheimer's disease, Down's Syndrome, amino, nitro, Calkoxy, Co alkylthio, and C hereditary cerebral hemorrhage with amyloidosis of the alkylcarboxyl, Dutch type, the amyloidosis of chronic inflammation, the amyloidosis of malignancy, familial Mediterranean fever, 0031 (c) aromatic and heteroaromatic groups Substituted with 2 or 3 adjacent hydroxy groups, multiple myeloma, B-cell dyscrasias, type II diabetes, the and optionally substituted with 1 to 5 non-inter prion diseases, Creutzfeldt-Jakob disease, Gerstmann fering Substituents, Straussler Syndrome, kuru, Scrapie, the amyloidosis associ ated with long-term hemodialysis, the amyloidosis associ 0032 (d) sugars, optionally substituted with one ated with carpal tunnel Syndrome, Senile cardiac or more anionic groupS. Selected from Sulfate, phosphate, phosphonate, carboxylate, and Sul amyloidosis, familial amyloidotic polyneuropathy, and the fonate groups, amyloidosis associated with endocrine tumors, and espe cially is Alzheimer's disease. 0033 (e) peptides and peptide derivatives, and 0034 (f) -C(O)R and -C(O)OR (where R is 0039. In a second aspect, this invention provides a drug Selected from the group consisting of (a) through product for the treatment of amyloidosis in a mammal (e) above); and Suffering therefrom, comprising a container labeled or accompanied by a label indicating that the drug product is 0035 Y is hydrogen, hydroxy, Calkoxy, benzy for the treatment of amyloidosis, the container containing loxy (where the phenyl group is optionally Substi one or more dosage units each comprising at least one tuted with 1 to 3 Substituents selected from halo and pharmaceutically acceptable excipient and, as an active Ce alkyl), or -OSOR (where R is C6, alkyl or phenyl optionally substituted with 1 to 3 substituents ingredient, an isolated pure compound Selected from those Selected from halo and C alkyl); used in the method of the first aspect of this invention. 0036 and the group of compounds consisting of 0040. In preferred embodiments of this second aspect, the acacetin, actinorhodine, alizarin, alizarin blue, drug product contains only one Such compound, the mam alizarin orange, alizarinSulfonic acid, alkannin, mal is a human; and the amyloidosis is Selected from the anthragallol, anthralin, anthrarobin, antharLfin, api group consisting of Alzheimer's disease, Down's Syndrome, genin, apigetirin, apiose, baicalein, baptigenin, 1.2, hereditary cerebral hemorrhage with amyloidosis of the 4-benzenetriol, boStrycoidin, carbidopa, canninic Dutch type, the amyloidosis of chronic inflammation, the acid, carubicin, cellobiose, centaurein, chloranilic amyloidosis of malignancy, familial Mediterranean fever, acid, chondrosine, chromotrope 2B, chromotropic multiple myeloma, B-cell dyscrasias, type II diabetes, the acid, chrySamminic acid, chrysarobin, chrysin, prion diseases, Creutzfeldt-Jakob disease, Gerstmann chrysophanic acid, cichoriin, citrazinic acid, citro Straussler Syndrome, kuru, Scrapie, the amyloidosis associ mycetin, collinomycin, curvularin, cyanidin, cyani ated with long-term hemodialysis, the amyloidosis associ din 3-glucoside, cyanidin 3-rhamnoglucoside, cya ated with carpal tunnel Syndrome, Senile cardiac nidin 3,5-diglucoside, cyanidin 3-Sophoroside, amyloidosis, familial amyloidotic polyneuropathy, and the daphnetin, datiscetin, daunorubicin, delphinidin, amyloidosis associated with endocrine tumors, and espe deoxyepinephrine, dioSmetin, dioSmin, dioxethe drine, dopa, dopamine, doxorubicin, droxidopa, cially is Alzheimer's disease. echinochrome A, embelin, emodin, ergoflavin, eri 0041. In a third aspect, this invention provides a method odictyol, esculetin, fenoldopam, fomecin A, fomecin of treating a disease characterized by C-Synuclein fibril B, fraxetin, fraxin, fredericamycin A, liunigatin, formation in a mammal Suffering therefrom, comprising fuSarubin, fuscin, fustin, galangin, gallein, gallocya nine, gardenin A, gardenin B, gardenin C, gardenin administration to the mammal of a therapeutically effective D, gardenin E, genistein, gentisin, granaticin, gua amount of an isolated pure compound Selected from the mecycline, hematein, hydroxySophorobioside, group consisting of the compounds of formula A, formula B, hydroxySophoricoside, icariin, isoquercitrin, formula C, formula D, and formula E: kaempferol, kermesic acid, laccaic acid A, laccaic acid B, laccaic acid C, laccaic acid D, leucocyanidin, luteolini, maclurin, menogaril, methylenedigallic Formula A acid, morin, oosporein, phenicin, phloroglucide, OH puberulic acid, puberulonic acid, purpurin, purpuro gallin, quercetagetin, quercimritrin, quinalizarin., quinic acid, resistomycin, rhamnetin, rhein, rhodi Zonic acid, rhodomycin A, rhodomycin B, robinin, ruberythric acid, rufigallol, rutin, Scutellarein, tannic acid, tetroquinone, tiron, troXerutin, and tunichrome B1, US 2001/0047032A1 Nov. 29, 2001

0049) (c) aromatic and heteroaromatic groups -continued Substituted with 2 or 3 adjacent hydroxy groups, Formula B and optionally substituted with 1 to 5 non-inter OH fering Substituents, R2 OH 0050 (d) sugars, optionally substituted with one or more anionic groups Selected from Sulfate, phosphate, phosphonate, carboxylate, and Sul R OH fonate groups, X 0051 (e) peptides and peptide derivatives, and Formula C R2 0.052 (f) –C(O)R and –C(O)OR (where R is Selected from the group consisting of (a) through Y. O X (e) above); and 0053 Y is hydrogen, hydroxy, C alkoxy, benzy Ri OH loxy (where the phenyl group is optionally Substi tuted with 1 to 3 Substituents selected from halo and OH O Ce alkyl), or -OSOR (where R is C alkyl or Formula D phenyl optionally substituted with 1 to 3 substituents Selected from halo and C alkyl); 0054 and the group of compounds consisting of acacetin, actinorhodine, alizarin, alizarin blue, alizarin orange, alizarinSulfonic acid, alkannin, anthragallol, anthralin, anthrarobin, antharufin, api genin, apigetirin, apiose, baicalein, baptigenin, 1.2, 4-benzenetriol, boStrycoidin, carbidopa, carminic acid, carubicin, cellobiose, centaurein, chloranilic acid, chondrosine, chromotrope 2B, chromotropic Formula E acid, chrySamminic acid, chrySarobin, chrysin, OH chrysophanic acid, cichorin, citrazinic acid, citro mycetin, collinomycin, curvularin, cyanidin, cyani OH din 3-glucoside, cyanidin 3-rhanuoglucoside, cyani din 3,5-diglucoside, cyanidin 3-Sophoroside, HO O daphnetin, datiscetin, daunorubicin, delphinidin, R deoxyepinephrine, dioSmetin, dioSmin, dioxethe drine, dopa, dopamine, doxorubicin, droxidopa, OR echinochrome A, embelin, emodin, ergoflavin., eri odictyol, esculetin, fenoldopam, fomecin A, fomecin OH B, fraxetin, fraxin, fredericamycin A, fumigatin, fuSarubin, fuscin, fustin, galangin, gallein, gallocya nine, gardenin A, gardenin B, grardenin C, gardenin 0042 where: D, gardenin E, genistein, gentisin, granaticin, gua mecycline, hematein, hydroxySophorobioside, 0043 R is selected from the group consisting of hydroxySophoricoside, icariin, isoquercitrin, hydrogen, 2,3-dihydroxybenzoyl, 3,4-dihydroxy kaempferol, kermesic acid, laccaic acid A, laccaic benzoyl, 2,34-trihydroxybenzoyl, and 3,4,5-trihy acid B, laccaic acid C, laccaic acid D, leucocyanidin, droxybenzoyl; luteolin, maclurin, menogaril, methylenedigallic acid, morin, oosporein, phenicin, phloroglucide, 0044) R' is hydrogen or OH: puberulic acid, puberulonic acid, purpurin, purpuro 0045 R and R are independently selected from gallin, quercetagetin, quercimritrin, quinalizarin, hydrogen and non-interfering Substituents, quinic acid, resistomycin, rhamnetin, rhein, rhodi Zonic acid, rhodomycin A, rhodomycin B, robinin, 0046 X is selected from hydrogen and the group ruberythric acid, rufigallol, rutin, Scutellarein, tannic consisting of acid, tetroquinone, tiron, troXerutin, and tunichrome B1, 0047) (a) hydroxy, amino, C, alkylamino, di(C, alkyl)anno, and cycloamino, 0055 and the pharmaceutically acceptable salts thereof. 0048 (b) C-2 alkyl, C-2 alkoxy, C-2 alky 0056. In preferred embodiments of this third aspect, only lthio, and C alkylcarboxyl, each optionally one Such compound is administered; the mammal is a Substituted with 1 to 5 moieties selected from the human; and the disease is Lewy body disease or Parkinson's group consisting of halogen, hydroxy, mercapto, disease, especially Parkinson's disease. amino, nitro, Calkoxy, Co alkylthio, and C 0057. In a fourth aspect, this invention provides a drug alkylcarboxyl, product for the treatment of a disease characterized by US 2001/0047032A1 Nov. 29, 2001

C-Synuclein fibril formation in a mammal Suffering there 0063 “Mammal” includes humans and non-human mam from, comprising a container labeled or accompanied by a mals, Such as companion animals (cats, dogs, and the like) label indicating that the drug product is for the treatment of and farm animals (cattle, horses, sheep,goats, Swine, and the a disease characterized by C-Synuclein fibril formation, the like). container containing one or more dosage units each com 0064. A “non-interfering substituent” is a substituent that, prising at least one pharmaceutically acceptable excipient when present in a compound, does not adversely affect the and, as an active ingredient, an isolated pure compound pharmacological activity of the compound and is not phar selected from those used in the method of the third aspect of maceutically undesirable. Suitable non-interfering Substitu this invention. ents include halogen and C alkyl and C alkoxy, each 0.058. In preferred embodiments of this fourth aspect, the optionally Substituted with up to 5 halogen atoms. drug product contains only one Such compound, the mam 0065 “Pharmaceutically acceptable excipient’ means an mal is a human; and the disease is Lewy body disease or excipient that is useful in preparing a pharmaceutical com Parkinson's disease, especially Parkinson's disease. position that is generally Safe, non-toxic, and desirable, and includes excipients that are acceptable for veterinary use as DETAILED DESCRIPTION OF THE well as for human pharmaceutical use. Such excipients may PREFERRED EMBODIMENTS be Solid, liquid, Semisolid, or, in the case of an aeroSol 0059) Definitions composition, gaseous. 0060 “Alkyl” means a linear hydrocarbyl group having 0066 “Pharmaceutically acceptable salts' means salts from one to the number of carbon atoms Specified, or a that are pharmaceutically acceptable and have the desired branched or cyclic hydrocarbyl group having from three to pharmacological properties. Such salts include Salts that the number of carbon atoms specified. “Alkyl” in this may be formed where acidic protons present in the com application is given a broader meaning than is conventional pounds are capable of reacting with inorganic or organic in organic chemistry and includes both Saturated groups bases. Suitable inorganic Salts include those formed with the (those conventionally known as alkyl groups), monounsat alkali metals, e.g. Sodium and potassium, magnesium, cal urated groups (Such as those conventionally known as cium, and aluminum. Suitable organic Salts include those alkenyl and alkynyl groups), and polyunsaturated groups, formed with organic baseS Such as the amine bases, e.g. except that the terms does not include groups containing ethanolamine, diethanolamine, triethanolamine, tromethar aromatic moieties, as the term "aromatic' is conventionally nine, N-methylglucamine, and the like. Such Salts also used. Exemplary C. alkyl groups include methyl, ethyl, include acid addition salts formed with inorganic acids (e.g. isopropyl, cyclopropyl, tert-butyl, cyclopropylmethyl, and hydrochloric and hydrobromic acids) and organic acids (e.g. hexyl. acetic acid, citric acid, maleic acid, and the alkane- and arene-Sulfonic acids Such as methaneSulfonic acid and ben 0061 An “aromatic' group is a cyclic (monocyclic, con Zenesulfonic acid). When there are two acidic groups densed bicyclic, or linked bicyclic) group having from 5 to present, a pharmaceutically acceptable Salt may be a mono 12 ring carbon atoms, and Sufficient ring unsaturation that the group is “aromatic” as that term is conventionally used. acid-mono-Salt or a di-Salt, and Similarly where there are Exemplary aromatic groups include phenyl, naphthyl, and more than two acidic groups present, Some or all of Such biphenylyl. A "heteroaromatic' group is an “aromatic' groups can be Salified. group as just defined in which from 1 to 4 of the ring carbon 0067. A “protecting group” has the meaning convention atoms have been replaced by O, S, or NR (where R is ally associated with it in organic Synthesis, i.e. a group that hydrogen or C. alkyl). Exemplary heteroaromatic groups Selectively blocks one or more reactive sites in a multifinc include pyrrolyl, furanyl, thiophenyl, benzofuranyl, indolyl, tional compound Such that a chemical reaction can be and the like. Such aromatic and heteroaromatic groups may carried out Selectively on another unprotected reactive site optionally be Substituted with 1 or more, especially 1 to 3, and Such that the group can readily be removed after the non-interfering Substituents. Selective reaction is complete. 0.062 An "isolated pure compound” is a compound in 0068 A “therapeutically effective amount’ means the isolated purified form Such as is conventional for active amount that, when administered to an animal for treating a ingredients in the pharmaceutical industry, and Specifically disease, is Sufficient to effect treatment for the disease. excludes the compound when found as a component in a “Treating” or “treatment” of the disease includes preventing mixture Such as within a plant or part thereof, or an extract the disease from occurring in a mammal that may be or decoction of Such plant or part, even when Such mixtures predisposed to the disease but does not yet experience or are partially purified to limit the number of components exhibit symptoms of the disease (prophylactic treatment), present therein. However, treatment with an "isolated pure inhibiting the disease (slowing or arresting its development), compound” is not limited to treatment with the compound providing relief from the Symptoms or side-effects of the alone but also includes treatment with the compound when disease (including palliative treatment), and relieving the present in a pharmaceutical composition of the type con disease (causing regression of the disease). "Treating amy ventional in pharmaceutical practice, i.e. including one or loidosis includes any one or more of the following: prevent more pharmaceutical excipients; however it specifically ing, inhibiting, reducing, disassembling, disrupting, and excludes treatment with the compound when the compound disaggregating amyloid fibrils and amyloid protein deposits, is found as a component in a mixture Such as within a plant such as AB and the other amyloids referred to in the or part thereof, or am extract or decoction of Such plant or BACKGROUND TO THE INVENTION. “Treating” an part, even when Such mixtures are partially purified to limit C-Synuclein fibril disease includes any one or more of the the number of components present therein. following: preventing, inhibiting, reducing, disassembling, US 2001/0047032A1 Nov. 29, 2001 disrupting, and disaggregating C-Synuclein fibrils and C-Sy (where the phenyl group is optionally Substituted nuclein-associated protein deposits, Such as those in Lewy with 1 to 3 substituents selected from halo and C. body disease and Parkinson's disease. alkyl and C alkoxy, each optionally Substituted 0069. The compounds found in the compositions and with 1 to 5 halogen atoms). used in the methods of this invention may possess one or 0081 and their individual stereoisomers, and the phar more chiral centers, and can therefore be produced as maceutically acceptable Salts thereof. individual Stereoisomers or as mixtures of Stereoisomers, depending on whether individual Stereoisomers or mixtures 0082 Preferred compounds include the compounds of of Stereoisomers of the Starting materials are used. Unless formula A and formula B, the compounds of formula C, the indicated otherwise, the description or naming of a com compounds of formula D, the compounds of formula E, and pound or group of compounds is intended to include both the the compounds of the list given following the descriptions of individual Stereoisomers or mixtures (racemic or otherwise) the formulae in the first aspect of the invention within the of stereoisomers. Methods for the determination of stere BRIEF SUMMARY OF THE INVENTION. ochemistry and the Separation of Stereoisomers are well 0083) A number of different preferences have been given known to a person of ordinary skill in the art see the above, and following any one of these preferences results in discussion in Chapter 4 of March J: Advanced Organic a compound or the composition or method of this invention Chemistry, 4th ed. John Wiley and Sons, New York, N.Y., that is more presently preferred than a compound in which 1992). that particular preference is not followed. However, these 0070 Presently Preferred Compounds preferences are generally independent and additive; and following more than one of these preferences may result in 0.071) While the broadest definition of the invention is set a more presently preferred compound than one in which out in the Summary of the Invention, certain compounds of fewer of the preferences are followed. this invention are presently preferred. 0084 Presently preferred compounds of this invention 0.072 Presently preferred compounds of this invention include 1,2,4-benzenetriol, ellagic acid, ethyl gallate, exi are compounds where: fone, gallamide, gallic acid, 5-hydroxydopamine, myricetin, 0073 R and Rare independently selected from the phloroglucide, propyl gallate, quercetin, quinic acid, and group consisting of hydrogen; C1-6 alkyl, (C-6 tannic acid. alkoxy, and C alkylthio (in each of which the alkyl 0085 Pharmacology and Utility group is optionally Substituted with 1 to 5 halogen atoms); and halo; 0086 The compounds of this invention act to inhibit or prevent amyloid fibril formation, inhibit or prevent amyloid 0074 X is selected from hydrogen and the group fibril growth, and/or cause disassembly, disruption, and/or consisting of disaggregation of preformed amyloid fibrils and amyloid 0075) (a) hydroxy, amino, C, alkylamino, di(C, protein deposits. Their activity can be measured in Vitro by alkyl)amino, and cycloamino, methods Such as those discussed in Examples 1 through 4 and ASSay 1 below, while their activity in Vivo against 0076 (b) C-2 alkyl, C-2 alkoxy, C-2 alky amyloidoses can be measured in animal models, Such as lthio, and C-2 alkylcarboxyl, each optionally those of Alzheimer's disease and in humans by a method Substituted with 1 to 5 moieties selected from the Such as that discussed in ASSay 2 below. group consisting of halogen, hydroxy, mercapto, amino, nitro, Calkoxy, Ce alkylthio, and C. 0087. The compounds of this invention also act to inhibit alkylcarboxyl, or prevent C.-synuclein fibril formation, inhibit or prevent C-Synuclein fibril growth, and/or cause disassembly, disrup 0077 (c) aromatic and heteroaromatic groups tion, and/or disaggregation of preformed C-Synuclein fibrils Substituted with 2 or 3 adjacent hydroxy groups, and C-Synuclein-associated protein deposits. Their activity and optionally substituted with 1 to 5 non-inter can be measured in vitro by methods Similar to those fering Substituents, and discussed in Examples 1 through 4 below. 0078 (d)-C(O)R and –C(O)OR (where R is 0088. The therapeutic ratio of a compound can be deter Selected from the group consisting of (a) through mined, for example, by comparing the dose that gives (c) above), effective anti-fibril (anti-amyloid or anti-C-Synuclein activ 0079 especially where X is selected from hydrogen ity in a Suitable in Vivo model in a Suitable animal Species and the group consisting of hydroxy, amino, Such as the mouse, with the dose that gives significant -C(O)R, and -C(O)OR (where R is selected weight loss (or other observable side-effects) in the test from hydroxy, amino, C alkyl optionally Substi animal Species. tuted with 1 to 5 halogen atoms, and aromatic and 0089 Pharmaceutical Compositions and Administration heteroaromatic groups Substituted with 2 or 3 adja cent hydroxy groups and optionally Substituted with 0090. In general, compounds of this invention will be 1 to 5 non-interfering Substituents Selected from administered in pure isolated form in therapeutically effec halogen atoms and C- alkyl and C- alkoxy, each tive amounts by any of the usual modes known in the art, optionally Substituted with 1 to 5 halogen atoms, and either Singly or in combination with at least one other compound of this invention and/or at least one other con 0080 Y is selected from the group consisting of ventional therapeutic agent for the disease being treated. A hydrogen, hydroxy, Ce alkoxy, and benzyloxy therapeutically effective amount may vary widely depending US 2001/0047032A1 Nov. 29, 2001

on the disease, its Severity, the age and relative health of the aqueous Suspensions. Such excipients are Suspending animal being treated, the potency of the compound(s), and agents, for example, Sodium carboxymethylcellulose, meth other factors. AS anti-fibril agents, therapeutically effective ylcellulose, hydroxypropylmethyl cellulose, Sodium algi amounts of compounds of this invention may range from nate, polyvinylpyrrolidone, gum tragacanth and gum acacia; 1-1000 mg/Kg body weight; for example, 10-100 mg/Kg. A dispersing or wetting agents may be naturally occurring person of ordinary skill in the art will be able without undue phosphatides, for example lecithin, or condensation prod experimentation, having regard to that Skill and this disclo ucts of an alkylene oxide with fatty acids, for example Sure, to determine a therapeutically effective amount of a polyoxyethylene Stearate, or condensation products of eth compound of this invention for the treatment of amyloidosis. ylene oxide with long chain aliphatic alcohols, for example, heptadecaethyleneoxycetanol, or condensation products of 0.091 In general, compounds of this invention will be ethylene oxide with partial esters derived from fatty acids administered as pharmaceutical compositions by one of the Such as hexitol Such as polyoxyethylene Sorbitol following routes: oral, topical, Systemic (e.g. transdermal, monooleate, or condensation products of ethylene oxide intranasal, or by Suppository), or parenteral (e.g. intramus with partial esters from fatty acids and a hexitol annhy cular, Subcutaneous, or intravenous injection). Compositions drides, for example, polyethylene Sorbitan monooleate. The may take the form of tablets, pills, capsules, Semisolids, aqueous Suspensions may also contain one or more preser powders, Sustained release formulations, Solutions, Suspen Vatives, for example, ethyl or n-propyl p-hydroxybenzoate, Sions, elixirs, aerosols, or any other appropriate composi one or more coloring agents, one or more flavoring agents, tions, and comprise at least one compound of this invention or one or more Sweetening agents, Such as Sucrose or in combination with at least one pharmaceutically accept Saccharin. able excipient. Suitable excipients are well known to per Sons of ordinary skill in the art, and they, and the methods 0095. Oily suspensions may be formulated by Suspending of formulating the compositions, may be found in Such the compound in a vegetable oil, for example arachis oil, standard references as Alfonso AR: Remington's Pharma olive oil, Sesame oil, or coconut oil or in a mineral oil Such ceutical Sciences, 17th ed., Mack Publishing Company, as liquid paraffin. The oily Suspensions may contain a Easton Pa., 1985. Suitable liquid carriers, especially for thickening agent, for example beeswax, hard paraffin or injectable Solutions, include water, acqueous Saline Solution, cetyl alcohol. Sweetening agents, Such as those Set forth aqueous dextrose Solution, and glycols. below, and flavoring agents may be added to provide a palatable oral preparation. These compositions may be pre 0092. In particular, the compound(s) preferably only Served by the addition of an antioxidant Such as ascorbic one Such compound is administered in any particular dosage acid. Dispersible powders and granules Suitable for prepa form-can be administered, orally, for example, as tablets, ration of an aqueous Suspension by the addition of water troches, lozenges, aqueous or oily Suspension, dispersible provide the active ingredient in admixture with a dispersing powders or granules, emulsions, hard or Soft capsules, or or Wetting agent, a Suspending agent and one or more Syrups or elixirs. Compositions intended for oral use may be preservatives. Suitable dispersing or wetting agents and prepared according to any method known in the art for the Suspending agents are exemplified by those already manufacture of pharmaceutical compositions and Such com described above. Additional excipients, for example Sweet positions may contain one or more agents Selected from the ening, flavoring and agents, may also be present. group consisting of Sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide 0096. The compounds may also be in the form of oil-in pharmaceutically elegant and palatable preparations. water emulsions. The oily phase may be a vegetable oil, for example olive oil or arachis oils, or a mineral oil, for 0.093 Tablets contain the compound in admixture with example liquid paraffin or mixtures of these. Suitable emul non-toxic pharmaceutically acceptable excipients which are Sifying agents may be naturally-occurring gums, for exa Suitable for the manufacture of tablets. These excipients may niple gum acacia or gum tragacanth, naturally occurring be for example, inert diluents, Such as calcium carbonate, phosphatides, for example Soybean, lecithin, and occurring Sodium carbonate, lactose, calcium phosphate or Sodium phosphatides, for example Soybean, lecithin, and esters or phosphate, granulating and disintegrating agents, for partial esters derived from fatty acids and hexitol anhy example, maize Starch or alginic acid; binding agents, for drides, for example Sorbitan monooleate, and condensation example, maize Starch, gelatin or acacia, and lubricating products of the Said partial esters with ethylene oxide, for agents, for example, magnesium Stearate or Stearic acid or example polyoxyethylene Sorbitan monooleate. The emul tale. The tablets may be uncoated or they may be coated by Sion may also contain Sweetening and flavoring agents. known techniques to delay disintegration and absorption in SyrupS and elixirs may be formulated with Sweetening the gastrointestinal tract and thereby provide a Sustained agents, for example, glycerol, Sorbitol or Sucrose. Such action over a longer period. For example, a time delay formulations may also contain a demulcent, a preservative material Such as glycerol monoStearate or glycerol distearate and flavoring and coloring agents. may be employed. Formulations for oral use may also be presented as hard gelatin capsules wherein the compound is 0097. The compound can also be administered by injec mixed with an inert Solid diluent, for example, calcium tion or infusion, either Subcutaneously or intravenously, or carbonate, calcium phosphate or kaolin, or as Soft gelatin intramuscularly, or intrastemally, or intranasally, or by infu capsules wherein the active ingredient is mixed with water Sion techniques in the form of Sterile injectable or oleagi or an oil medium, for example, peanut oil, liquid paraffin or nous Suspension. The compound may be in the form of a olive oil. Sterile injectable aqueous or oleaginous Suspensions. These Suspensions may be formulated according to the known art 0094. Aqueous suspensions contain the compound in using Suitable dispersing of wetting agents and Suspending admixture with excipients Suitable for the manufacture of agents which have been described above. The sterile inject US 2001/0047032A1 Nov. 29, 2001 able preparation may also be a sterile injectable Solution or protection of the hydroxy groups that form an essential part Suspension in a non-toxic parenterally-acceptable diluent or of the compounds, and the knowledge and use of Such Solvent for example, as a Solution in 1,3-butanediol. Among protecting groups will be within the knowledge of a perSon the acceptable vehicles and Solvents that may be employed of ordinary skill in the art. are water, Ringer's Solution and isotonic Sodium chloride 0103) The starting materials, intermediates, and com Solution. In addition, Sterile, fixed oils are conventionally pounds of this invention may be isolated and purified using employed as a Solvent or Suspending medium. For this conventional techniques, including filtration, distillation, purpose any bland fixed oils may be conventionally crystallization, chromatography, and the like. They may be employed including Synthetic mono- or diglycerides. In characterized using conventional methods, including physi addition fatty acids Such as oleic acid find use in the cal constants and Spectral data. preparation of injectables. 0.098 Dosage regimens can be adjusted to provide the EXAMPLES optimum therapeutic response. For example, Several divided 0104. The following non-limiting examples illustrate the dosages may be administered daily or the dosage may be invention. proportionally reduced as indicated by the exigencies of the therapeutic Situation. Example 1 0099] It is especially advantageous to formulate the com 0105 Disassembly/Disruption of Alzheimer's Disease pounds in dosage unit form for ease of administration and A? 1-42 Fibrils by Polyhydroxylated Aromatic Compounds uniformity of dosage. Dosage unit form as used herein refers 0106. In this study, different types of commercially avail to physically discrete units Suited as unitary dosages for the able compounds which consist of various polyhydroxylated Subjects to be treated; each containing a therapeutically aromatic containing Structures were tested for their ability to effective quantity of the compound and at least one phar cause a disassembly/disruption of pre-formed Alzheimer's maceutical excipient. A drug product will comprise a dosage disease amyloid fibrils containing AB 1-42. This type of unit form within a container that is labeled or accompanied activity would be important for any potential anti-amyloid by a label indicating the intended method of treatment, Such drug which can be used in patients who already have as the treatment of an amyloid disease, Such as Alzheimer's Substantial amyloid deposition in organs and/or tissues. For disease, or of a disease associated with C-Synuclein fibril example, Alzheimer's disease patients in mid-to-late Stage formation, Such as Parkinson's disease. A “therapeutically disease have abundant AB-containing amyloid deposits in effective dosage” preferably inhibits amyloidosis or a dis their brains as part of both neuritic plaques and cerebrovas ease associated with C-synuclein fibril formation in a patient cular amyloid deposits. A compound capable of causing by at least 20, more preferably by at least 40%, even more disassembly/disruption of pre-existing amyloid deposits preferably by at least 60%, and still more preferably by at would be advantageous for use in these patients who are at least 80%, relative to untreated subjects. latter Stages of the disease process. 0100 Preparation of the Compounds of this Invention 0107 For the first study, 1 mg of AB 1-42 (Bachem Inc., Torrance, Calif., USA) was dissolved in 1.0 ml of double 0101 Many of the compounds used in the compositions distilled water (1 mg/ml solution). 25 uM of AB 1-42 was and methods of this invention are well known to the art. then incubated overnight (-18 hours) at 37 C., in the They may be briefly described in such references as the absence or presence of 100 ug/ml of the following com Merck Index, 12th edition, Merck & Co., Inc., Whitehouse pounds: 1) EDTA (Sigma Chemical Company, St. Louis, Station, N.J., 1996 (which typically provides a reference to Mo., USA), 2) myricetin (Acros, Somerville, N.J., USA), 3) exifone (Acros) 4) pyrogallol (Sigma), 5) tannic acid a synthesis or isolation), and may be found in chemical (Acros), 6) pyrocatechol (Acros), 7) quercetin (Sigma), 8) catalogs, Such as those of commercial SupplierS Such as ellagic acid (Acros), 9) 1,2,4-benzenetriol (Acros), 10) 5-hy Aldrich Chemical Company (Milwaukee, Wis.), Bachem droxydopamine (Acros), 11) gallamide hydrate (AcroS), 12) (Torrance, Calif.), Sigmna (St. Louis, Mo.). gallic acid (Sigma), 13) ethyl gallate (Acros), 14) quinic acid 0102) For those compounds that are novel, the starting (Acros), 15) propyl gallate (Sigma), and 16) phloroglucide materials and reagents used in preparing these compounds (Acros), each in the presence of 150 min Tris HCI, 10 mM are generally available from commercial SupplierS Such as NaCl (pH 7.0) with 0.02% sodium azide. In this study, the Aldrich Chemical Company, Bachem, and Sigmna, or are AD 1-42:compound weight ratio was 1:1. prepared by methods well known to a perSon of ordinary 0108) For the second study, 1 mg of AB 1-42 (Bachem) skill in the art following procedures described in Such was dissolved in 1.0 ml of double distilled water (1 mg/ml references as Fieser and Fieser's Reagents for Organic solution). 25 uM of AB 1-42 was then incubated overnight Synthesis, Vols 1-17, John Wiley and Sons, New York, N.Y., (-18 hours) at 37 C., in the absence or presence of 50 tug/ml 1991; Rodd's Chemistry of Carbon Compounds, vols. 1-5 of the following compounds: 1) gallic acid, 2) ethyl gallate, and supps, Elsevier Science Publishers, 1989; Organic Reac 3) quinic acid, 4) gallamide trihydrate, 5) ellagic acid, 6) tions, vols 1-40, John Wiley and Sons, New York, NY, 1991; propyl gallate, and 7) pyrogallol, each in the presence of 150 March J: Advanced Organic Chemistry, 4th ed. John Wiley mM Tris HCl, 10 mM NaCl (pH 7.0) with 0.02% sodium and Sons, New York, NY, 1992; and Larock: Comprehensive azide. In this study, the AB 1-42:compound weight ratio was Organic Transformations, VCH Publishers, 1989, and the 2:1. Syntheses of the novel compounds will be readily Suggested 0109) A previously described method of measuring amy to a perSon or ordinary skill in the art by reference to known loid fibril formation utilizing Thioflavin T fluorometry (H analogs (such as the commercially available analogs referred Naiki et al., Lab. Invest. 65:104-110, 1991; H Levine III, to above) of the novel compounds. Many Such preparations Protein Sci. 2:404-410, 1993; H Levine III, Amyloid: Int. J will involve the use of protecting groups, especially for the Exp. Clin. Invest. 2:1-6, 1995; H Naiki and K. Nakakuki, US 2001/0047032A1 Nov. 29, 2001

Lab. Invest. 74:374-383, 1996) was employed to identify potential therapeutic compounds capable of causing a dis TABLE 1-continued assembly/disruption of Alzheimer's AB 1-42 amyloid fibrils. Thioflavin T is known to bind to fibrillar amyloid proteins, Disassembly/disruption of Alzheimer's 1-42 fibrils, and an increase in fluorescence correlates with an increase in as indicated by Thioflavin T fluorescence inhibition amyloid fibril formation, whereas a decrease in fluorescence Fluorescence inhibition, %, at the correlates with a decrease in amyloid fibril due to disassem A? 1-42: compound bly and/or disruption. The Alzheimer's AB protein (1-42) W/w ratios given when placed in Solution, Such as distilled water, tends to Spontaneously form amyloid fibrils. Using this Sensitive Compound name 1:1 2:1 assay, any decreases or increases in fluorescence was pre Gallic acid 57 1.9 44 - 1.7 viously shown to correlate with a decrease or increase in the Ethyl gallate 490.8 3O3.7 amount of amyloid fibrils (see the documents cited above), Quinic acid 31 - 9.0 OS 3.9 allowing one to identify and quantitate the extent of poten Phloroglucide 3O 0.6 tial inhibitors and/or enhancers of Alzheimer's AB 1-42 Propyl gallate 29 2.8 38 - 4.8 amyloid fibrils. 0110. To assess the effects of each compound on potential 0113 EDTA, a known chelating agent, caused no signifi disassembly/disruption of preformed AB 1-42 fibrils, 50 ul cant disassembly/disruption of AB 1-42 amyloid fibrils, of AB 1-42 with or without test compounds (described Suggesting that the inhibitory effects observed with polyhy above) were added to 1.2 ml of 100 uM ThioflavinT (Sigma) in 50 mM NaH2PO (pH 6.0) for fluorometry readings. droxylated aromatic compounds was not attributable to their Studies indicated that increasing concentrations of A3 gave ability to complex metals. a proportional increase in fluorescence in the presence of 1 00 M Thioflavin T, ruling out the presence of any dispro Example 2 portionate inner filter effects in these Studies. Fluorescence emission at 482 nm was measured on a Turner instrument 0114 Dose-Dependent Disassembly/Disruption of model 450 fluorometer at an excitation wavelength of 450 Alzheimer's Disease AB 1-40 Fibrils by Tannic Acid and um. For each determination, the fluorometer was calibrated Gallic Aid by Zeroing in the presence of the Thioflavin Treagent alone, 0.115. In this study, the potential dose-dependent effects and by setting the 50 ng/ml riboflavin (Sigma Chemical Co., of tannic acid and gallic acid on disassembly/disruption of St. Louis, Mo.) in the Thioflavin Treagent to 1800 fluores cence units. All fluorescence determinations were based on pre-formed AB 1-40 was assessed. In this experiment, 1 mg these references and any fluorescence given off by any of the of AB 1-40 (Bachem Inc., Torrance, Calif., USA; Lot compounds in the presence of the Thioflavin Treagent was #T-20824) was dissolved in 1.0 ml of double distilled water always Subtracted from all pertinent readings. (1 rmg/ml solution) and incubated for 4 days at 37° C. to spontaneously induce fibril formation. 25 uM of pre 0111 For all fibrillogenesis studies utilizing Thioflavin T fibrillized AB 1-40 was then incubated overnight (-18 hours) fluorometry, as disclosed herein, comparisons of amyloid at 37 C., in the absence or presence of increasing amounts protein in the presence or absence of test compounds were (25 ug/ml, 50 ug/ml, 75 ug/ml and 100 ug/ml) of tannic acid based on paired Student's t tests with data shown as the or gallic acid (each in the presence of 150 mM Tris HCl, 10 mean of triplicate measurements:tstandard deviation. mM NaCl, pH 7.0, with 0.02% sodium azide). The 0112 As shown in Table 1, the polyhydroxylated aro AB:compound weight ratioS were therefore 4:1, 2:1, 4:3, and matic compounds caused a disassembly/disruption of AB 1:1, respectively. 50 plaliquots were then added to 1.2 ml of 1-42 amyloid fibril as determined by inhibition of Thioflavin 100 uM Thioflavin T (Sigma) in 50 mM NaH2PO (pH 6.0) T fluorescence. All results were significant at the p <0.005 for fluorometry readings as described in Example 1 above. level, except that for quinic acid at the 2:1 ratio (asterisked 0116. As shown in Table 2, both tannic acid and gallic in Table 1), which was not significant. acid caused a dose-dependent disassembly/disruption of Af 1-40 amyloid fibrils as indicated by a dose-dependent inhi TABLE 1. bition of Thioflavin T fluorescence. All results were signifi Disassembly/disruption of Alzheimer's 1-42 fibrils, cant at the p<0.005 level, except that for gallic acid at the 4:1 as indicated by Thioflavin T fluorescence inhibition ratio (asterisked in Table 2), which was significant at the p-0.05 level. Fluorescence inhibition, %, at the A? 1-42: compound W/w ratios given TABLE 2 Compound name 1:1 2:1 Dose-dependent disassembly/disruption of Alzheimer's 1-40 fibrils, as indicated by Thioflavin T fluorescence inhibition Myricetin 94 O.9 Exifone 93 - 1.4 Fuorescence inhibition, %, Pyrogallol 89 6.7 72 3.8 at the Afs 1-40: compound Tannic acid 77 1.3 WFW ratios given Pyrocatechol 77 2.6 Quercetin 76 0.6 Compound name 4:1 2:1 4:3 1:1 Ellagic acid 74 1.4 62 - 39 1,2,4-Benzenetriol 71 - 3.3 Tannic acid 31 4.8 42 + 2.8 493.7 53 + 4.2 5-Hydroxydopamine 70 1.1 Gallic acid 148.2 22 - 3.3 34 it 3.6 45 - 4.1 Gallamide trihydrate 65 - 12.3 60 - 22 US 2001/0047032A1 Nov. 29, 2001

Example 3 TABLE 3 0117 Disaggregation of Alzheimer's Disease AB 1-40 Fibrils by Polyhydroxylated Aromatic Compounds Disaggregation of Alzheimer's 1-40 fibrils, as indicated by Congo red spectrophotometry 0118. In this study, a Congo red-AB spectrophotometric Compound name Decrease in absorbance, 26 assay (Klunk et al., Anal. Biochem. 266:66-76, 1999) was Gallic acid 52 + 0.4 modified to determine the effectiveness of polyhydroxylated Ethyl gallate 28, 5.0 aromatic compounds on the disaggregation of pre-formed Quinic acid O 5.0** A? 1-40 amyloid fibrils. For this assay, 1 mg of AB 1-40 Gallamide trihydrate 31 - 1.9 Ellagic acid 54 - 2.8 (Bachem) was incubated for 4 days in distilled water at 37 Propyl gallate 17 - 7.3* C. to spontaneously produce amyloid fibrils. 25 uM of Pyrogallol 63 - 3.4 fibrillized AB 1-40 was then incubated in triplicate with various test compounds for 3 days at 37 C. in Tris-buffered saline (TBS)(100 mM Tris; 50 mM NaCl; pH 7.0, with Example 4 0.02% sodium azide), at an AB:compound weight ratio of 2:1. Following incubation, 50 ul of 360 uM Congo red 0.122 Dose-Dependent Disaggregation of Alzheimer's (Sigma) in distilled water was then added to 250 ul of each Disease AB 1-40 Fibrils by Tannic Acid and Gallic Acid incubation mixture, giving a final AB:Congo red molar ratio 0123. In this study, the potential dose-dependent effects of 1:3. After 10 minutes, the absorbance at 405 nm (refer of tannic acid and gallic acid on the disaggregation of ence wavelength to account for the absorbance of Congo red fibrillized AB 1-40 was assessed. In this experiment, the modified Congo red-A? spectrophotometric assay (Klunket alone at 540 nm) and 540 nm (sample absorbance where al, Anal. Biochem. 266:66-76, 1999) was used as described “Sample” refers to AD alone, test compound alone, or AB above (i.e. Example 4). However, in this specific experiment plus test compound, all in the presence of Congo red) was increasing amounts of tannic acid or gallic acid (i.e. 25 determined using a Biorad Model 550 ELISA Plate Reader Aug/ml, 50 tug/ml, 75 ug/ml and 100 ug/ml) were tested (Biorad, Hercules, Calif., USA). The absorbance at wave following an overnight (-18 hours) incubation at 37° C. in length 405 nm was automatically subtracted by the ELISA the presence of 25uM of AB 1-40 (Bachem). plate reader from the absorbance at wavelength 540 nm 0124. As shown in Table 4, both tannic acid and gallic (difference is referred to as A absorbance) (see Klunk et al. acid caused a dose-dependent disaggregation of AB 1-40 cited above). Therefore, the Aabsorbance reading at 540 nm amyloid fibril as determined by decreases in Thioflavin T was proportional to the amount of aggregated AB left in fluorescence. All results were significant at the p<0.001 Solution (Klunlc et al.). level, except that for gallic acid at the 4:1 ratio (asterisked 0119 For all experiments involving test compounds, the in Table 4), which was significant at the p<0.05 level. Aabsorbance reading at 540 nrrL of the test compound alone TABLE 4 (in the absence of Af), was always Subtracted from the Dose-dependent disaggregation of Alzheimer's 1-40 fibrils, corresponding A absorbance reading at 540 nm of the test as indicated by Congo red spectrophotometry compound in the presence of AB. Using this modification of Decrease in absorbance, 26, the method of Klunk et al., the use of a greater final at the Afs 1-42: compound concentration of Congo red, i.e. 60 uM instead of 141 uM, in W/w ratios given the presence of fibrillar AB gave an overall absorbance at Compound name 4:1 2:1 4:3 1:1 540 nm that was always below 1.0 Absorbance Unit (AU), Tannic acid 42 + 5.2 48 - 6.8 59 - 6.4 61 + 11.1 and well within the linear absorbance range. Gallic acid 179.5* 22 - 4.0 30 - 6.0 32 4.8 0120) The following polyhydroxylated aromatic contain ing compounds were tested using the above described Example 5 Congo red-AB Spectrophotometric assay to determine their effectiveness on disaggregation of pre-formed AB 1-40 0.125 Disassembly/Disruption of Islet Amyloid Ribrils amyloid fibrils: 1) gallic acid, 2) ethyl gallate, 3) quinic acid, (Amylin) by Polyhydroxylaited Aromatic Compounds 4) gallamide trihydrate, 5) elagic acid, 6) propyl gallate, and 0.126 90% of patients with type II diabetes demonstrate 7) pyrogallol. the deposition and accumulation of amyloid fibrils in the islets of Langerhans in the pancreas (Cooper et al., Proc. 0121 The polyhydroxylated aromatic compounds had Natl. Acad. Sci. USA 84:8628-8632, 1987). This amyloid Varying effects on causing disaggregation of pre-aggregated protein involved consists of a 37 amino acid protein known as islet amyloid polypeptide or amylin. Islet amyloid is A? 1-40 amyloid fibrils as determined using the Congo red believed to contribute to the destruction of the beta-cells of Spectrophotometric assay described above. The results were the pancreas, thus eventually leading many patients to Significant at the p<0.005 level, except for propyl gallate become insulin-dependent (i.e. type I diabetes). Amylin has (asterisked in Table 3) at the p<0.05 level, and quinic acid the ability to also form Substantial amyloid fibrils immedi (double asterisked), which was not significant. ately when placed in Solution. The next Study was therefore US 2001/0047032A1 Nov. 29, 2001

implemented to determine whether Some of the Specific concentration of 25-50 uM. After 4 days of treatment, polyhydroxylated aromatic containing compounds which Viability is assessed by measurement of lactate dehydroge cause a disassembly/disruption of AB fibrils, also cause a nase (LDH) released into culture medium. Lactate dehydro disassembly/disruption of islet amyloid fibrils. genase (LDH) is measured in 20 gl aliquots of conditioned defined DMEM using a standard 340 nm kinetic LDH assay 0127. For this study, the method of Thioflavin T fluo (Sigma Catalog Number #228-20) in a 96 well format. rometry as described in Example 1 was used. Briefly, 25 uM Assays are performed at 37 C. in a PC-driven EL340 of human amylin (Bachem) was incubated overnight (-18 Microplate Biokinetics plate reader (Bio-Tek Instruments) hours) at 37 C., alone or in the presence of 100 lug/ml of the using Delta Soft II software (v. 3.30B, BioMetallics, Inc.) following compounds: 1) exifone, 2) myricetin, and 3) for data analysis. Quality control Standards containing nor tannic acid, each in the presence of 150 mM Tris HCl, 10 maland elevated levels of serum LDH (for example, Signma mM NaCl, pH 7.0, with 0.02% sodium azide, at an amylin Enzyme Controls 2N and 2E) are run with every assay. :compound weight ratio of 1:1. Results are expressed as units of LDII/L where 1 unit is 0128. Following Thioflavin T fluorometry readings as defined as the amount of enzyme that will catalyze the described in Example 1, 5 ul aliquots of amylin only, formation of 1 micromole of nicotinamide adenine dinucle amylin+myricetin, amylin+exifone, and amylin+tannic acid otide per minute under conditions of the assay. For protec were also taken, allowed to air dry overnight on gelatin tion Studies, a compound of formula 1 is added to cultures coated Slides, and Stained with Congo red as previously prior to and/or concurrently with the amyloid-beta treat described (Castillo et al., Diabetes 47:612-620, 1998). ment. 0129. As shown in Table 5, the polyhydroxylated aro 0.135 Activity of the compounds is illustrated by a matic compounds which were very effective in causing a decrease in LDH released into the media (a neurotoxic disassembly/disruption of AB 1-42 amyloid fibrils were also indicator), as compared to control. effective in causing a disassembly/disruption of islet amy loid fibrils. All results were significant at the p-0.005 level. 0136 Assay 2 0.137 Five to fifty women are selected for a clinical study. TABLE 5 The Women are post-menopausal, i.e., have ceased menstru ating for between 6 and 12 months prior to the study’s Disassembly/disruption of amylin fibrils, initiation, have been diagnosed with early Stage Alzheimer's as indicated by Thioflavin T fluorescence inhibition disease (AD), are expected to have worsening Symptoms of Compound name Fluorescence inhibition, % AD within the study period, but are in good general health Myricetin 97.4 O.3 otherwise. The Study has a placebo control group, i.e., the Exifone 99.1 - 0.5 Women are divided into two groups, one of which receives Tannic acid 83.8 - 1.4 the compound of this invention and the other receives a placebo. The patients are benchmarked as to memory, cog nition, reasoning, and other Symptoms associated with AD. 0130 Congo red staining experiments confirmed the dis Women in the test group receive a therapeutic dose of the assembly/disruption of amylin fibrils by polyhydroxylated compound per day by the oral route. They continue this aromatic compounds initially demonstrated by Thioflavin T therapy for 6-36 months. Accurate records are kept as to the fluorometry Studies as described above. Congo red Staining benchmarked Symptoms in both groupS and at the end of the of amylin alone demonstrated positive staining (i.e. classic Study these results are compared. The results are compared red/green birefringence as viewed under polarized light and both between members of each group and also the results for indicative of amyloid) (Puchtler et al., J. Histochem. each patient are compared to the Symptoms reported by each Cytochem. 10:355-364, 1962). In comparison, an overnight patient before the Study began. Activity of the compound is incubation with exifone, myricetin or tannic acid resulted in illustrated by an attenuation of the typical cognitive decline a marked decrease in Congo red Staining, Suggestive of an and/or behavioral disruptions associated with AD. amylin fibril disassembly/disruption. 0.138. Utility of the compounds is evidenced by activity 0131 Further in vitro and in vivo assays may be used to in at least one of the above assayS. test the compounds for their effectiveness in the treatment of 0.139 While this invention has been described in con Alzheimer's disease, Such as those described in European junction with Specific embodiments and examples, it will be Published Patent Application No. 0 659 418. apparent to a perSon of ordinary skill in the art, having 0132) Stock solutions of peptides (1 mM) are freshly regard to this disclosure, -that equivalents of the Specifically prepared in pyrogen-free Sterile water and diluted to the disclosed materials and techniques will also be applicable to indicated concentrations in defined culture media. Rat hip this invention; and Such equivalents are intended to be pocampal cultures (10-14 days in vitro) are treated with included within the following claims. peptides or vehicle for four days. The viability of the rat cortical cultures is visually assessed by phase contrast microScopy and quantified by measuring lactate dehydroge What is claimed is: nas, (LDH) released into the culture media. 1. A method of treating amyloidosis in a mammal Suffer ing therefrom, comprising acdministration to the mammal of 0133) Assay 1 a therapeutically effective amount of an isolated pure com 0134) Primary rat hippocampal neurons are cultured in pound Selected from the group consisting of the compounds Vitro with Standard cell culture techniques. Amyloid-beta of formula A, formula B, formula C, formula D, and formula (AB) peptide is added to cultured cells at a normally toxic E: US 2001/0047032A1 Nov. 29, 2001 13

5 moieties Selected from the group consisting of halogen, hydroxy, mercapto, anino, nitro, C Formula A alkoxy, Ce alkylthio, and C alkylcarboxyl, OH (c) aromatic and heteroaromatic groups Substituted R2 OH with 2 or 3 adjacent hydroxy groups, and optionally substituted with 1 to 5 non-interfering Substituents, (d) Sugars, optionally Substituted with one or more X OH anionic groups Selected from Sulfate, phosphate, R1 phosphonate, carboxylate, and Sulfonate groups, Formula B (e) peptides and peptide derivatives, and OH (f) -C(O)R and -C(O)OR (where R is selected R2 OH from the group consisting of (a) through (e) above); and Y is hydrogen, hydroxy, C alkoxy, benzyloxy (where R OH the phenyl group is optionally Substituted with 1 to 3 X Substituents selected from halo and C alkyl), or Formula C -OSOR4 (where R4 is C alkyl or phenyl option R2 ally substituted with 1 to 3 substituents selected from Y. O X halo and CI6 alkyl); and the group of compounds consisting of acacetin, actinorhodine, alizarin, alizarin blue, alizarin orange, Ri OH alizarinSulfonic acid, alkannin, anthragallol, anthralin, anthrarobin, antharu.fm, apigenin, apigetirin, apiose, OH O baicalein, baptigenin, 1,2,4-benzenetriol, boStrycoidin, Formula D carbidopa, carminic acid, carubicin, cellobiose, centau OH rein, chloranilic acid, chondrosine, chromotrope 2B, chromotropic acid, chrySamminic acid, chrySarobin, chrysin, chrysophanic acid, cichorin, citrazinic acid, citromycetin, collinomycin, curvularin, cyanidin, cya nidin 3-glucoside, cyanidin 3-rhamnoglucoside, cyani din 3,5-diglucoside, cyanidin 3-Sophoroside, daphne tin, clatiscetin, daunorubicin, delphinidin, deoxyepinephrine, dioSmetin, dioSmin, dioxethedrine, dopa, dopamine, doxorubicin, droxidopa, echi Formula E nochrome A, embelin, emodin, ergoflavin, eriodictyol, OH eSculletin, fenoldopam, fomecin A, fomecin B, fraxetin, fraxin, fredericamycin A, fumigatin, fuSarubin, fuscin, OH fustin, galangin, gallein, gallocyanine, gardenin A, gar denin B, gardenin C, gardenin D, gardenin E, genistein, HO O gentisin, granaticin, guamecycline, hematein, hydrox R ySophorobioside, hydroxySophoricoside, icariin, iso quercitrin, kaempferol, kermesic acid, laccaic acid A, laccaic acid B, laccaic acid C, laccaic acid D, leuco OR cyanidin, luteolin, maclurin, menogaril, methylenedi OH gallic acid, morin, oosporein, phenicin, phloroglucide, puberulic acid, puberulonic acid, purpurin, purpurogal where: lin, quercetagetin, quercinritrin, quinalizarin, quinic acid, resistomycin, rhamnetin, rhein, rhodizonic acid, R is Selected from the group consisting of hydrogen, rhodomycin A, rhodomycin B, roblinin, ruberythric 2,3-dihydroxybenzoyl, 3,4-dihydroxybenzoyl, 2,34-tri acid, rufigallol, rutin, Scutellarein, tannic acid, tetro hydroxybenzoyl, and 3,4,5-trihydroxybenzoyl; quinone, tiron, troXerutin, and tunichrome B 1, R" is hydrogen or OH; but excluding pyrogallol, R" and R2 are independently selected from hydrogen and and the pharmaceutically acceptable Salts thereof. non-interfering Substituents, 2. The method of claim 1 where only one such compound X is Selected from hydrogen and the group consisting of is administered. 3. The method of claim 1 where the mammal is a human. (a) hydroxy, amino, CI6 alkylamino, di(CI6 alky 4. The method of claim 3 where the amyloidosis is l)amino, and cycloamino, Selected from the group of diseases consisting of Alzhe (b) C1-22 alkyl, C-22 alkoxy, C1-22 alkylthio, and C1-22 imer's disease, Down's Syndrome, hereditary cerebral hem alkylcarboxyl, each optionally substituted with 1 to orrhage with amyloidosis of the Dutch type, the amyloidosis US 2001/0047032A1 Nov. 29, 2001 of chronic inflammation, the amyloidosis of malignancy and familial Mediterranean fever, the amyloidosis of multiple -continued myeloma and B-cell dyscrasias, the amyloidosis of type II Formula E diabetes, the amyloidosis of the prion diseases, Creutzfeldt OH Jakob disease, Gerstnamnn-Straussler Syndrome, kuru, OH Scrapie, the amyloidosis associated with long-term hemodi alysis, the amyloidosis associated with carpal tunnel Syn HO O drome, Senile cardiac amyloidosis, familial amyloidotic R polyneuropathy, and the amyloidosis associated with endo crine tumors. OR

5. The method of claim 4 where the amyloidosis is OH Alzheimer's disease. 6. A drug product for the treatment of amyloidosis in a mammal Suffering therefrom, comprising a container labeled where: or accompanied by a label indicating that the drug product R is Selected from the group consisting of hydrogen, is for the treatment of amyloidosis, the container containing 2,3-dihydroxybenzoyl, 3,4-dihydroxybenzoyl, 2,34-tri one or more dosage units each comprising at least one hydroxybenzoyl, and 3,4,5-trihydroxybenzoyl; pharmaceutically acceptable excipient and, as an active R" is hydrogen or OH; ingredient, an isolated pure compound Selected from the group consisting of the compounds of formula A, formula B, R" and R2 are independently selected from hydrogen and formula C, formula D, and formula E: non-interfering Substituents, X is Selected from hydrogen and the group consisting of (a) hydroxy, amino, Ce alkylamino, di(C16 alky l)amino, and cycloamino, Formula A OH (b) C-2 alkyl, C-2 alkoxy, C-2 alkylthio, and Cl alkylcarboxyl, each optionally substituted with 1 to R2 OH 5 moieties selected from the group consisting of halogen, hydroxy, mercapto, amino, nitro, C alkoxy, Ce alkylthio, and C alkylcarboxyl, X OH (c) aromatic and heteroaromatic groups Substituted R1 with 2 or 3 adjacent hydroxy groups, and optionally Formula B substituted with 1 to 5 non-interfering Substituents, OH (d) Sugars, optionally Substituted with one or more R2 OH anionic groups Selected from Sulfate, phosphate, phosphonate, carboxylate, and Sulfonate groups, (e) peptides and peptide derivatives, and Ri OH (f) -C(O)R and -C(O)OR (where R is selected X from the group consisting of (a) through (e) above); Formula C and R2 Y is hydrogen, hydroxy, C alkoxy, benzyloxy (where Y. O X the phenyl group is optionally Substituted with 1 to 3 Substituents selected from halo and C alkyl), or -OSOR4 (where R4 is Cl(, alkyl or phenyl optionally R OH Substituted with 1 to 3 Substituents selected from halo and C-6 alkyl); OH O Formula D and the group of compounds consisting of acacetin,

OH actinorhodine, alizarin, alizarin blue, alizarin orange, alizarinSulfonic acid, alkannin, anthragallol, anthralin, anthrarobin, antharutfin, apigenin, apigetirin, apiose, baicalein, baptigenin, 1,2,4-benzenetriol, boStrycoidin, carbidopa, carminic acid, carubicin, cellobiose, centau rein, chloranilic acid, chondrosine, chromotrope 2B, chromotropic acid, chrysanminic acid, chrySarobin, chrysin, chrysophanic acid, cichorin, citrazinic acid, citromycetin, collinomycin, curvularin, cyanidin, cya nidin 3-glucoside, cyanidin 3-rhamnoglucoside, cyani din 3,5-diglucoside, cyanidin 3-Sophoroside, daphne US 2001/0047032A1 Nov. 29, 2001 15

tin, datiscetin, daunorubicin, delphinidin, deoxyepinephrine, dioSmetin, dioSmin, dioxethedrine, -continued Formula D dopa, doparnine, doxorubicin, droxidopa, echi nochrome A, embelin, emodin, ergoflavin, eriodictyol, eSculletin, fenoldopam, fomecin A, fomecin B, fraxetin, fraxin, fredericamycin A, fumigatin, fuSarubin, fuscin, fustin, galangin, gallein, gallocyanine, gardenin A, gar denin B, gardenin C, gardenin D, gardenin E, genistein, gentisin, granaticin, guamecycline, hematein, hydroxySophorobioside, hydroxySophorico Side, icariin, isoquercitrin, kaempferol, kermesic acid, laccaic acid A, laccaic acid B, laccaic acid C, laccaic acid D, leucocyanidin, luteolin, maclurin, menogaril, Formula E methylenedigallic acid, morin, Oosporein, phenicin, OH phloroglucide, puberulic acid, puberulonic acid, pur purin, purpurogallin, quercetagetin, quercinritrin, OH quinalizarin, quinic acid, resistomycin, rhamnetin, HO rhein, rhodizonic acid, rhodomycin A, rhodomycin B, R robinin, ruberythric acid, rufigallol, rutin, Scutellarein, tannic acid, tetroquinone, tiron, tron erutin, and tunichrome B1, OR but excluding pyrogallol, OH

and the pharmaceutically acceptable Salts thereof. where: 7. The drug product of claim 5 containing only one Such R is Selected from the group consisting of hydrogen, compound. 2,3-dihydroxybenzoyl, 3,4-dihydroxybenzoyl, 2,34-tri 8. The drug product of claim 7 indicated for the treatment hydroxybenzoyl, and 3,4,5-trihydroxybenzoyl; of Alzheimer's disease. 9. A method of treating a mammal Suffering from a R" is hydrogen or OH: disease characterized by C-Synuclein fibril formation, com R and R2 are independently Selected from hydrogen and prising administration to the mammal of a therapeutically non-interfering Substituents, effective amount of an isolated pure compound Selected from the group consisting of the compounds of formula A, X is Selected from hydrogen and the group consisting of formida B, formula C, formula D, and formula E: (a) hydroxy, amino, Ce alkylamino, di(C. alky l)amino, and cycloamino, (b) C-2 alkyl, C-2 alkoxy, Cl2 alkylthio, and C-2 Formula A alkylcarboxyl, each optional Ly Substituted with 1 to OH 5 moieties Selected from the group consisting of R2 OH halogen, hydroxy, mercapto, amino, nitro, C alkoxy, C alkylthio, and C alkylcarboxyl, (c) aromatic and heteroaromatic groups Substituted X OH with 2 or 3 adjacent hydroxy groups, and optionally R1 substituted with 1 to 5 non-interfering Substituents, Formula B (d) Sugars, optionally Substituted with one or more OH anionic groups Selected from Sulfate, phosphate, R2 OH phosphonate, carboxylate, and Sulfonate groups, (e) peptides and peptide derivatives, and (f) -C(O)R and -C(O)OR (where R is selected Ri OH from the group consisting of (a) through (e) above); X and Formula C R2 Y is hydrogen, hydroxy, C alkoxy, benzyloxy (where the phenyl group is optionally Substituted with 1 to 3 Y. O X Substituents selected from halo and C alkyl), or -OSOR4 (where R4 is C alkyl or phenyl option ally substituted with 1 to 3 substituents selected from R OH halo and C alkyl); OH O and the group of compounds consisting of acacetin, actinorhodine, alizarin, alizarin blue, alizarin orange, US 2001/0047032A1 Nov. 29, 2001 16

alizarinSulfonic acid, alkannin, anthragallol, anthralin, anthrarobin, anthani?in, apigenin, apigetirin, apiose, -continued baicalein, baptigenin, 1,2,4-benzenetriol, boStrycoidin, Formula B carbidopa, carminic acid, carubicin, cellobiose, centau OH rein, chloranilic acid, chondrosine, chromotrope 2B, chromotropic acid, chrySamminic acid, chrySarobin, R2 OH chrysin, chrysophanic acid, cichorin, citrazinic acid, citromycetin, collinomycin, curvularin, cyanidin, cya nidin 3-glucoside, cyanidin 3-rhamnoglucoside, cyani R OH din 3,5-diglucoside, cyanidin 3-Sophoroside, daphne tin, diatiscetin, daunorubicin, delphinidin, Formula C deoxyepinephrine, dioSmetin, dioSmin, dioxethedrine, R2 cLopa, dopamine, doxorubicin, droxidopa, echi nochrome A, embelin, emodin, ergoflavin, eriodictyol, Y. O X eSculletin, fenoldopam, fomecin A, fomecin B, fraxetin, fraxin, fredericamycin A, iumigatin, fuSarubin, fuscin, fustin, galangin, gallein, gallocyanine, gardenin A, gar Ri OH denin B, gardenin C, gardenin D, gardenin E, genistein, OH O gentisin, granaticin, guamecycline, hematein, hydrox Formula D ySophorobioside, hydroxySophoricoside, icariin, iso quercitrin, kaempferol, kermesic acid, laccaic acid A, laccaic acid B, laccaic acid C, laccaic acid D, leuco cyanidin, luteolin, maclurin, menogaril, methylenedi gallic acid, morin, oosporein, phenicin, phloroglucide, puberulic acid, puberulonic acid, purpurin, purpurogal lin, quercetagetin, quercimritrin, quinalizarin, quinic acid, resistomycin, rhamnetin, rhein, rhodizonic acid, rhodomycin A, rhodomycin B, rob3inin, ruberythric acid, rufigallol, rutin, Scutellarein, tannic acid, tetro quinone, tiron, troXerutin, and tunichrome B1, Formula E OH but excluding pyrogallol, OH and the pharmaceutically acceptable Salts thereof. 10. The method of claim 9 where only one such com HO O pound is administered. R 11. The method of claim 10 where the mammal is a human. OR 12. The method of claim 11 where the disease is Lewy body disease or Parkinson's disease. OH 13. The method of claim 12 where the disease is Parkin Son's disease. where: 14. A drug product for the treatment of a disease charac terized by C-synuclein fibril formation in a mammal suffer R is Selected from the group consisting of hydrogen, ing therefrom, comprising a container labeled or accompa 2,3-dihydroxybenzoyl, 3,4-dihydroxybenzoyl, 2,34-tri nied by a label indicating that the drug product is for the hydroxybenzoyl, and 3,4,5-trihydroxybenzoyl; treatment of a disease characterized by o-synuclein fibril R" is hydrogen or OH; formation, the container containing one or more dosage R" and R2 are independently selected from hydrogen and units each comprising at least one pharmaceuticFLlly non-interfering Substituents, acceptable excipient and, as an active ingredient, an isolated pure compound Selected from the group consisting of the X is Selected from hydrogen and the group consisting of compounds of formula A, formula B, formula C, formula D, (a) hydroxy, amino, C alkylamino, di(Cl alky and formula E: l)amino, and cycloamino, (b) C1-22 alkyl, C-22 alkoxy, C1-22 alkylthio, and C1-22 alkylcarboxyl, each optionally substituted with 1 to Formula A 5 moieties Selected from the group consisting of OH halogen, hydroxy, mercapto, amino, nitro, C OH alkoxy, Clo alkylthio, and Cl 6 alkylcarboxyl, (c) aromatic and heteroaromatic groups Substituted with 2 or 3 adjacent hydroxy groups, and optionally OH substituted with 1 to 5 non-interfering Substituents, (d) Sugars, optionally Substituted with one or more anionic groups Selected from Sulfate, phosphate, phosphonate, carboxylate, and Sulfonate groups, US 2001/0047032A1 Nov. 29, 2001 17

(e) peptides and peptide derivatives, and hydroxy, mercapto, amino, nitro, Co alkoxy, C alkylthio, and Clo alkylcarboxyl, (f) -C(O)R and -C(O)OR (where R is selected from the group consisting of (a) through (e) above); (c) aromatic and heteroaromatic groups Substituted with 2 and or 3 adjacent hydroxy groups, and optionally Substi tuted with 1 to 5 non-interfering Substituents, and Y is hydrogen, hydroxy, Clo alkoxy, benzyloxy (where the phenyl group is optionally Substituted with 1 to 3 (d)-C(O)R and -C(O)OR (where R is selected from Substituents selected from halo and CI alkyl), or the group consisting of (a) through (c) above). -OSOR4 (where R4 is CI 6alkyl or phenyl optionally 19. The method of claim 1 where X is selected from Substituted with 1 to 3 Substituents selected from halo hydrogen and the group consisting of hydroxy, amino, and C alkyl); and the group of compounds consisting -C(O)R, and -C(O)OR (where R is selected from of acacetin, actinorhodine, alizarin, alizarin blue, hydroxy, amino, C , alkyl optionally Substituted with 1 to alizarin orange, alizarinSulfonic acid, alkannin, 5 halogen atoms, and aromatic and heteroaromatic groups anthragallol, anthralin, anthrarobin, anthanifin, apige Substituted with 2 or 3 adjacent hydroxy groups and option nin, apigetrin, apiose, baicalein, baptigenin, 1,2,4-ben ally substituted with 1 to 5 non-interfering substituents Zenetriol, boStrycoidin, carbidopa, carminic acid, caru Selected from halogen atoms and Cl6 alkyl and C alkoxy, bicin, cellobiose, centaurein, chloranilic acid, each optionally Substituted with 1 to 5 halogen atoms. chondrosine, chromotrope 2B1, chromotropic acid, 20. The method of claim 1 where Y is selected from the chrySamminic acid, chrySarobin, chrysin, chrysophanic group consisting of hydrogen, hydroxy, Clo alkoxy, and acid, cichorin, citrazinic acid, citromycetin, collino benzyloxy (where the phenyl group is optionally Substituted mycin, curvularin, cyanidin, cyanidin 3-glucoside, cya with 1 to 3 substituents selected from halo and C alkyl and nidin 3-rhamnoglucoside, cyanidin 3,5-diglucoside, C. alkoxy, each optionally Substituted with 1 to 5 halogen cyanidin 3-Sophoroside, daphnetin, datiscetin, dauno atoms). rubicin, delphinidin, deoxyepinephrine, dioSmetin, 21. The method of claim 1 where the compound is a dioSmin, dioxethedrine, cIlopa, dopamine, doxorubi compound of formula A or formula B, or a pharmaceutically cin, droxidopa, echinochrome A, embelin, emodin, acceptable Salt thereof. ergoflavin, eriodictyol, esculetin, fenoldopam, fomecin 22. The method of claim 21 where the compound is A, fomecin B, fraxetin, fraxin, fredericamycin A, Selected from the group consisting of dibromogallic acid, i-umigatin, uffsarubin, fuscin, fustin, galangin, gallein, digallic acid, ethyl gallate, exifone, fisetin, gallacetophe gallocyanine, gardenin A, gardenin B, gBardenin C, none, gallami,de, gallic acid, C-glucogallin, fB-glucogallin, gardenin D, gardenin E, genistein, gentisin, granaticin, 5-hydroxydopamine, and propyl gallate, and the pharma guamecycline, hematein, hydroxySophorobioside, ceutically acceptable Salts thereof. hydroxySophoricoside, icariin, isoquercitrin, 23. The method of claim 1 where the compound is a kaempferol, kermesic acid, laccaic acid A, laccaic acid compound of formula C or a pharmaceutically acceptable B, laccaic acid C, laccaic acid D, leucocyanidin, luteo Salt thereof. lin, maclurin, menogaril, methylenedigallic acid, 24. The method of claim 22 where the compound is morin, oosporein, phenicin, phloroglucide, puberulic Selected from the group consisting of myricetin and quer acid, puberulonic acid, purpurin, purpurogallin, querc cetin, and the pharmaceutically acceptable Salts thereof. etagetin, quercimritrin, quinalizarin, quinic acid, resis 25. The method of claim 1 where the compound is a tomycin, rhamnetin, rhein, rhodizonic acid, rhodomy compound of formula D or a pharmaceutically acceptable cin A, rhodomycin B, robinin, ruberythric acid, Salt thereof. rafigallol, rutin, Scutellarein, tannic acid, tetroquinone, 26. The method of claim 25 where the compound is tiron, troXeirutin, and tunichrome B1, ellagic acid or a pharmaceutically acceptable Salt thereof. 27. The method of claim 1 where the compound is a but excluding pyrogallol, compound of formula E or a pharmaceutically acceptable and the pharmaceutically acceptable Salts thereof. Salt thereof. 15. The drug product of claim 14 containing only one Such 28. The method of claim 27 where the compound is compound. Selected from the group consisting of catechin, epicatechin, 16. The drug product of claim 15 indicated for the gallocatechin, epigallocatechin, and their gallate esters, and treatment of Parkinson's disease. the pharmaceutically acceptable Salts thereof. 17. The method of claim 1 where R' and R are indepen 29. The method of claim 1 where the active ingredient is dently Selected from the group consisting of hydrogen; C. Selected from group of compounds consisting of acacetin, actinorhodine, alizarin, alizarin blue, alizarin orange, alizar alkyl, Calkoxy, and C alkylthio (in each of which the insulfonic acid, alkannin, anthragallol, anthralin, anthrar alkyl group is optionally Substituted with 1 to 5 halogen obin, antharufin, apigenin, apigetirin, apiose, baicalein, bap atoms); and halo. tigenin, 1 2,4-benzenetriol, boStrycoidin, carbidopa, 18. The method of claim 1 where X is selected from carminic acid, carubicin, cellobiose, centaurein, chloranilic hydrogen and the group consisting of acid, chondrosine, chromotrope 2B, chromotropic acid, (a) hydroxy, amino, Calkylamino, di(Calkyl)amino, chrySamminic acid, chrySarobin, chrysin, chrysophanic acid, and cycloamino, cichorin, citrazinic acid, citromycetin, collinomycin, cur Vularin, cyanidin, cyanidin 3-glucoside, cyanidin 3-rhamno (b) C1-22 alkyl, C-22 alkoxy, C1-22 alkylthio, and C1-22 glucoside, cyanidin 3,5-diglucoside, cyanidin 3-Sophoro alkylcarboxyl, each optionally substituted with 1 to 5 Side, daphnetin, datiscetin, daunorubicin, delphinidin, moieties Selected from the group consisting of halogen, deoxyepinephrine, dioSmetin, dioSmin, dioxethedrine, dopa, US 2001/0047032A1 Nov. 29, 2001 dopamine, doxorubicin, droxidopa, echinochrome A, embe quinic acid, resistomycin, rhamnetin, rhein, rhodizonic acid, lin, emodin, ergoflavin, eriodictyol, esculetin, fenoldopam, rhodomycin A, rhodomycin B, robinin, ruberythric acid, fomecin A, fomecin B, fraxetin, fraxin, fredericamycin A, rufigallol, rutin, Scutellarein, tannic acid, tetroquinone, tiron, fumigatin, fuSarubin, fuscin, fustin, galangin, gallein, gal troXerutin, and tunichrome B 1, and the pharmaceutically locyanine, gardenin A, gardenin B, gardenin C, gardenin D, acceptable Salts thereof. gardenin E, genistein, gentisin, granaticin, guamecycline, 30. The method of claim 1 where the compound is hematein, hydroxySophorobioside, hydroxySophoricoside, Selected from 1,2,4-benzenetriol, elagic acid, ethyl gallate, icariin, isoquercitrin, kaempferol, kermesic acid, laccaic exifone, gallamide, gallic acid, 5-hydroxydopamine, acid A, laccaic acid B, laccaic acid C, laccaic acid D, myricetin, phloroglucide, propyl gallate, quercetin, quinic leucocyanidin, luteolin, maclurin, menogaril, methylenedi acid, and tannic acid, and the pharmaceutically acceptable gallic acid, morin, oosporein, phenicin, phloroglucide, salts thereof. puberulic acid, puberulonic acid, purpurin, purpurogallin, pyrocatechol, quercetagetin, quercimritrin, quinalizarin,