Inhibitors of microbial-induced amyloid

Abstract

To provide compounds useful for prevention of amyloid formation and treatment of amyloid-related disorders including synucleinopathies such as Parkinson's disease.SOLUTION: A compound of formula (I) or a pharmaceutically acceptable salt thereof: SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation of U.S. Provisional Patent Application No. USSN 62, filed January 10, 2020. / 959,385, claiming priority under 35 U.S.C. § 119(e). , the entire contents of which are incorporated herein by reference.

[0002] The present disclosure relates to inhibitors of amyloid formation, particularly inhibitors of microbial-induced amyloid formation, and to treat or inhibit neurological and other disorders associated with amyloid accumulation. The present invention relates to methods of using inhibitors of amyloid formation. A method is also provided. [Background technology]

[0003] Many neurodegenerative diseases are associated with atypical aggregation of proteins in the brain, which This leads to cell death, resulting in a variety of neurological disorders. (ii) the specific proteins that affect the brain; and (iii) the specific areas of the brain affected. This is thought to be the result of specific neuronal cell types undergoing α-synuclein synthesis. In the case of alpha-synucleinopathies, abnormal aggregation of this protein is responsible for over 50 "alpha-synucleinopathies." This can result in one of several disorders, of which Parkinson's disease is the most common and most widely studied. In Parkinson's disease, α-synuclein aggregation causes damage to certain nerve cells. Lewy bodies are formed within the neuronal cell types, most typically those that produce the neurotransmitter dopamine. When sufficient α-synuclein aggregates are present, the nuclei accumulate as large precipitates. Neuronal death occurs, leading to a decrease in dopamine production. Dopamine is necessary for proper control of movement. Once dopaminergic neurons die, they do not come back. , the dopamine pool is irreversibly reduced until motor symptoms progress and become debilitating.

[0004] The most pathogenic form of α-synuclein remains unknown, e.g., complete α-synuclein Intact Lewy bodies or smaller oligomeric α-synuclein fibrils contribute to disease progression It is unclear whether this is closely related to the pathology of α-synuclein. Traditional pharmaceutical and biotechnological approaches to treat disease symptoms have It focuses on attacking aggregation processes in neurons and brain regions that are also involved in Small molecule, antibody and vaccine approaches are all being investigated. It continues to be tried and evaluated as an intervention for Alzheimer's disease and other α-synucleinopathies. Importantly, all of these strategies currently have the ability to cross the blood-brain barrier and reach target neural tissues. The penetration of the blood-brain barrier remains a key challenge for the treatment of neurodegenerative diseases. This is one of the most important pharmacokinetic issues hindering drug development. Inhibitors of amyloid formation that may provide therapeutic benefit without the need to cross the brain barrier, particularly There is a need for α-synuclein aggregation inhibitors. Summary of the Invention

[0005] In one aspect, a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , R 4 , L 1 , L 2 , A1 , A 2 , X, Y and Z are as defined herein. Provided herein are: [ka]

[0006] In some embodiments, the compound of formula (I) is a compound of formula (II), (III), (IV), A structure selected from (V), (VI), (VII), (VIII), (IX), and (X). It may have a structure. [ka]

[0007] The compounds disclosed herein are available in the form of compositions formulated for delivery outside the systemic circulation of a subject. The compositions may be formulated, for example, for enteral or intranasal delivery. and / or the composition may be further formulated for controlled release in the lower intestine or colon. In some embodiments, the composition can be administered to a subject by oral administration, injection (e.g., intravenous, subcutaneous, intramuscular, intraperitoneal, intraspinal or intracranial), topical delivery, mucosal delivery, or administration to the central nervous system or The compositions may be formulated for delivery to the systemic circulation. Capsules, tablets, soft gels, spray-dried powders, polymer matrices, hydrogels, enteric coated coated solids, crystalline solids, amorphous solids, glassy solids, coated particulates The composition may comprise a powder, liquid, spray liquid, aerosol or microcapsules.

[0008] The present disclosure further provides a method for disrupting the formation of amyloid aggregates, comprising administering to a subject amyloid or amyloid-containing compound. a method comprising contacting a precursor of amyloid with a compound or composition thereof described herein. Provide the law.

[0009] The present disclosure further provides a method of disrupting the formation of amyloid aggregates in a subject, the method comprising administering to a subject a therapeutically effective amount of amyloid aggregates ... The method includes administering to the subject a compound described herein or a composition thereof. Optionally, the subject further receives a drug to disrupt the formation of amyloid aggregates prior to administration of the composition. Such selection or identification is carried out in accordance with the methods described above. Before administering the composition, a clinical or diagnostic evaluation may be performed. The subjects included Parkinson's disease, dementia with Lewy bodies, incident Lewy body disease, and Alzheimer's disease. Lewy body variant of Immer's disease, multiple system atrophy, or pure autonomic failure, or any of these Optionally, administration of the composition The prevention or inhibition of amyloid aggregate formation in the subject is measured before, during, or after administration. It is determined or evaluated.

[0010] The present disclosure further provides a method for inhibiting, ameliorating, or potentially improving amyloid disorders in a subject in need thereof. A method for reducing the likelihood of, delaying the onset of, treating or preventing a disease comprising administering to a subject a compound described herein. Optionally, the subject may further comprise administering to the subject a compound or composition thereof. Additionally, for example, the presence or absence of bacterial proteins (such as CsgA) in a subject's intestinal sample may be monitored. detects the level, or the presence or level of a microorganism that produces the bacterial protein. a compound that inhibits or prevents the formation of amyloid aggregates prior to administration of the composition. Such selection or identification may be carried out by selecting or identifying the compound as a compound that will benefit from the or before administering the composition, by clinical or diagnostic evaluation. The subjects selected were Parkinson's disease, dementia with Lewy bodies, incident Lewy body disease, and Alzheimer's disease. Lewy body variant of Zheimer's disease, multiple system atrophy, or pure autonomic failure, or any of these Optionally, the compound or or inhibiting the formation of amyloid aggregates in the subject before, during, or after administration of the composition. The harm or interference is measured or assessed.

[0011] In some embodiments, the amyloid aggregates are formed from one or more mammalian proteins, such as proteins. Animal amyloid or mammalian amyloid precursors, and / or one or more bacterial amyloid precursors or a fungal protein (e.g., a composition comprising CsgA). In some embodiments, the amyloid protein may be one or more mammalian proteins, such as proteins. mammalian amyloid or mammalian amyloid precursor, and / or one or more bacteria Some compositions may contain bacterial or fungal proteins (e.g., compositions containing CsgA). In embodiments, the amyloid aggregates are located in the gastrointestinal tract, enteric nerve tissue, dural sinuses, oral cavity, or In some embodiments, the amyloid protein may be present in the nasal cavity (e.g., the olfactory bulb). Proteins are present in the gastrointestinal tract, enteric nerve tissue, dural sinuses, oral cavity, or nasal cavity (e.g., olfactory bulb). obtain.

[0012] In some embodiments, the methods of the present disclosure are used to treat, for example, anosmia, dysosmia, bradykinesia, Ataxia, tremor, muscle rigidity, posture and balance disorders, loss of motility, dysarthria or other symptoms Changes in speech, changes in handwriting, orthostatic hypotension, memory problems, swallowing problems, incontinence, sleep problems, cardiac arrhythmias psychiatric symptoms, including visual disturbances, depression, and / or visual, auditory, olfactory, and / or tactile hallucinations Problems, dizziness, cognitive impairment, changes in dopamine levels, changes in serotonin levels, a subject, which may include one or more of: a change in renin levels; measuring or assessing changes in the nervous system of a subject, e.g., neurological symptoms, motor behavior, or other behavior of the subject The method further includes:

[0013] In some embodiments, the methods of the present disclosure involve treating changes in the gastrointestinal system, e.g., gastrointestinal symptoms in a subject. The method further includes measuring or evaluating the characteristics of the gastrointestinal system, and the changes in the gastrointestinal system may include, for example, dysphagia, intestinal Decreased motility, gastroparesis, constipation (including chronic constipation and chronic idiopathic constipation), small intestinal bacterial abnormalities overgrowth (SIBO), diarrhea, abdominal pain and / or cramps, bloating, flatulence, nausea, or irritable bowel syndrome Any other symptoms of IBS, inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease , increased intestinal permeability, leaky gut, intestinal dysbiosis, excessive salivation (sialorrhea), anus may include one or more of rectal dysfunction, bowel dysfunction, or any combination thereof Such hyperpermeability is due to inflammation of the intestinal lining and / or failure of tight junctions between cells of the intestinal epithelium. This may result in material passing from the lumen into the surrounding tissue, some of which may be This leakage of material from the digestive tract or intestinal lumen can enter the peritoneal cavity and / or the systemic circulation. Intestinal hyperpermeability is sometimes referred to as "leaky gut" or "leaky gut syndrome." be.

[0014] In some embodiments, the compounds and compositions of the present disclosure may be used to treat a range of conditions, including those that present with a neurological symptom or condition. In some embodiments, the compositions of the present disclosure can be administered to a subject before or after the onset of The composition is administered to a subject before or after the onset of gastrointestinal symptoms or conditions associated with an amyloid disorder. In some embodiments, the subject may be administered a steroid drug, e.g., after a clinical or diagnostic evaluation. Depending on the level, it can be classified as dementia with Lewy bodies, paraneoplastic Lewy body disease, or Alzheimer's disease. Small body variant, multiple system atrophy, pure autonomic failure, or any combination of these. selected as those identified as being at risk or already having the condition. In terms of form, the above subjects are under 18 years old, 18-30 years old, 30-50 years old, 50-60 years old, 60 In some embodiments, the subject is clinically or Patients who are at risk for or have been diagnosed with Parkinson's disease through diagnostic evaluation or family history analysis, etc. The subject is already identified or selected as having.

[0015] In some embodiments, the compounds or compositions described herein may contain caffeine, niacin, or niacin. Cotine, theophylline, theobromine, xanthine, methylxanthine, or derivatives thereof In some embodiments, the methods disclosed herein can be co-administered with a conductor. The method further comprises administering to the subject an α-synuclein aggregation inhibitor. In embodiments, the methods disclosed herein involve administering to the subject L-DOPA, carbodopa, or levothyroxine. Vodopa, droxidopa, rasagiline, apomorphine hydrochloride, bromocriptine, rotigotine pramipexole, ropinirole, benztropine, trihexyphenidyl, selegiline Phosphorus, entacapone, tolcapone, amantadine, pimabanelcin, rivastigmine, etc. In some embodiments, the method further comprises administering to a subject a subject, the method comprising administering to a subject a subject a therapeutically effective amount of a compound selected from the group consisting of benzodiazepines, ... The methods disclosed herein include administering to the subject an alpha-synuclein aggregation inhibitor. Contains L-DOPA, carbodopa, levodopa, droxidopa, rasagiline, apomorphine methicillin hydrochloride, bromocriptine, rotigotine, pramipexole, ropinirole, benzothiazol-1 lopine, trihexyphenidyl, selegiline, entacapone, tolcapone, amantadine , pimabanelcin, rivastigmine, or any combination thereof, is administered to the subject. In some embodiments, the method further comprises administering to a subject a compound comprising an α-synuclein aggregation inhibitor and a L -DOPA, carbodopa, levodopa, droxidopa, rasagiline, apomorphine hydrochloride, Bromocriptine, rotigotine, pramipexole, ropinirole, benztropine, Lihexyphenidyl, selegiline, entacapone, tolcapone, amantadine, pimabane In some embodiments, the leucine, rivastigmine, etc. are administered in the same composition. Alpha-synuclein aggregation inhibitors and L-DOPA, carbodopa, levodopa, droxidopa, Rasagiline, apomorphine hydrochloride, bromocriptine, rotigotine, pramipexole, Ropinirole, benztropine, trihexyphenidyl, selegiline, entacapone, It is administered in a separate composition from lucapone, amantadine, pimabanelcin, rivastigmine, etc. In some embodiments, the separate compositions are administered simultaneously. In this embodiment, the separate compositions are administered at different times.

[0016] In some embodiments, the compounds or compositions described herein are for medical use. In some embodiments, the compositions described herein are administered to patients with an amyloid disorder described herein. (such as an amyloid disorder in Table 2). So, amyloid disorders include Parkinson's disease, dementia with Lewy bodies, and incident Lewy body disease. Alzheimer's disease with Lewy bodies, multiple system atrophy, or pure autonomic failure, or In some embodiments, the present invention is selected from the group consisting of any combination of two or more thereof. The compounds or compositions described herein may be used to treat amyloid disorders described herein (see Table 2). The invention relates to the preparation of a medicament for the treatment of certain disorders, such as myeloathene-associated neuromyelopathy. In embodiments, the amyloid disorder is Parkinson's disease, dementia with Lewy bodies, incident Lewy disease, Alzheimer's disease with Lewy bodies, the Lewy body variant of Alzheimer's disease, multiple system atrophy, or pure autonomic dysfunction or any combination of two or more thereof. In embodiments, the compositions include one or more compounds of the invention described herein. In this embodiment, the composition is formulated for delivery to the gastrointestinal tract, e.g., by oral or rectal delivery. In some embodiments, the composition is formulated with an enteric coating. The composition may be formulated for delivery to the central nervous system, e.g., by intraspinal or intracranial delivery. In some embodiments, the composition is formulated to cross the blood-brain barrier. Such formulations are formulated to bypass the blood-brain barrier, e.g., by intranasal administration. Such formulations can also be administered via the olfactory route.

[0017] The present disclosure identifies compounds or compositions that affect the formation of microbially induced amyloid. In some approaches, the methods provide methods for treating a patient with a compound or composition described herein. In the presence of the composition, multiple concentrations of microbial amyloid or microbial amyloid precursor are a concentration of alpha-synuclein and / or other mammalian amyloid or mammalian amyloid and analyzing or measuring the formation of amyloid after said reaction. and comparing said analysis or measurement with a control analysis or measurement, analyzing or measuring amyloid formation after the reaction in the absence of the composition. In some methods and compositions disclosed herein, the microbial amyloid or In some embodiments, the microbial amyloid precursor comprises CsgA. The method may also include agitation during contacting and / or prior to measuring.

[0018] In certain embodiments, the above-mentioned multiple concentrations of microbial amyloid or microbial amyloid a composition containing CsgA at multiple concentrations of α-synuclein and / or Contacting with other mammalian amyloids or mammalian amyloid precursors may result in the formation of amyloids. In some further embodiments, the indicator is , fluorescent indicators, spin-label indicators, enzymes, antibodies, or colorimetric indicators. In another embodiment, the indicator is thioflavin T (ThT). When the indicator is an antibody, the method of the present disclosure further comprises detecting the presence of the antibody in the aggregated alpha-synuclein. and / or specific for another mammalian amyloid or mammalian amyloid precursor and optionally a fluorescent label, an enzyme, a colorimetric label, a spin label, a metal ion binding moiety , electrochemiluminescent labels, nucleic acids, polysaccharides or polypeptides. In some embodiments of the method, CsgA and the alpha-synuclein and / or other Such bacterial amyloid precursors and / or mammalian amyloids / mammalian amyloids The ide precursors are each separately labeled.

[0019] In some embodiments according to the methods of the present disclosure, the presence of a compound or composition described herein In the presence of multiple concentrations of microbial amyloid or microbial amyloid precursor (e.g., Cs a composition containing α-synuclein and / or other mammalian amyloids at multiple concentrations and contacting the mammalian amyloid precursor with the amyloid precursor, and forming amyloid after the reaction. and comparing said analysis or measurement with a control analysis or measurement. altering or modulating amyloid formation or It further includes identifying or selecting compounds or compositions suspected of causing the In embodiments, the methods described herein involve administering to a subject a compound that reduces or enhances amyloid formation. In some embodiments, the method further comprises identifying or selecting a composition as described herein. The described methods involve administering compounds or The compounds or compositions identified by these methods are further included in the identification or selection of compounds. The composition may comprise a compound that alters amyloid formation (e.g., a compound that preferably crosses the blood-brain barrier). identified as a population that would benefit from administration of a compound that reduces amyloid formation without or a selected subject. Such a selected subject may be a patient suffering from Parkinson's disease. , Dementia with Lewy bodies, Incidental Lewy body disease, Lewy body variant of Alzheimer's disease Diagnosis or diagnosis may be made as multiple system atrophy, or pure autonomic failure, or any combination of these. may have been evaluated.

[0020] The methods according to the present disclosure include the step of administering a compound or composition described herein in the presence or absence of the compound or composition. Several concentrations of microbial amyloid or microbial amyloid precursor were combined with several concentrations of α-synuclein. Contact with rhein and / or other mammalian amyloid or mammalian amyloid precursor causing the microorganism to produce amyloid; and Further contemplated are methods for producing microbially-induced amyloid, including analyzing or quantifying the amyloid. In some further embodiments, the microbial amyloid or pre-microbial amyloid In some further embodiments, the method according to the present disclosure comprises the step of: In some further embodiments, the method further comprises stirring during contact or before measurement. In some further embodiments, the assay is performed in the presence of an indicator of amyloid formation. The indicator of dode formation may include a fluorescent indicator, a spin label indicator, or a colorimetric indicator. In some embodiments, the indicator is thioflavin T (ThT). In the form of CsgA and α-synuclein, or other such bacterial amyloids / bacteria mammalian amyloid precursor and mammalian amyloid / mammalian amyloid precursor, respectively. In some embodiments, the amyloid formation is detected by intrinsic fluorescence, dyes, or other methods. or fluorescence of the label, fluorescence resonance energy transfer, fluorescence polarization, fluorescence polarization transfer, UV / Vis spectroscopy Optical methods, magnetic resonance, Raman scattering, electron paramagnetic spin resonance, optical microscopy, electron microscopy, chemotaxis The surface roughness is analyzed or measured by scanning tunneling microscopy or atomic force microscopy.

[0021] In some embodiments according to the methods of the present disclosure, multiple concentrations of microbial amyloid or microorganisms are A biological amyloid precursor (e.g., a composition containing CsgA) is mixed with multiple concentrations of α-synucleic acid. and / or other mammalian amyloid or mammalian amyloid precursor. The composition present during the treatment comprises a mixture of compounds. In some embodiments, the composition may comprise extracts of feces, urine, blood, marrow, or the like. In some embodiments, the composition comprises natural fluid, saliva, or components thereof. In some further embodiments, the natural product comprises an extract from an herb. In some embodiments, the natural product is a fungal tissue, a plant material, or a food product. legumes, seeds, berries, leaves, fruits, flowers, plant roots, plant stems or plant bark. In some embodiments, the composition comprises one or more bacteria, bacterial extracts, lysates, conditioned culture media, or the like. freeze-dried bacteria, freeze-dried lysate, freeze-dried culture medium, or any combination thereof In some embodiments, the composition may comprise one or more microorganisms, a microbial extract, or a combination thereof. products, lysates, conditioned culture media, lyophilized microorganisms, lyophilized lysates, lyophilized culture media In some embodiments, the method may include: , a composition that increases or decreases amyloid formation, preferably a compound that does not cross the blood-brain barrier. Compounds identified by these methods may be used in a variety of applications. Compounds that alter amyloid formation (e.g., amyloid-binding proteins) are preferably administered without crossing the blood-brain barrier. identified or selected as a population that would benefit from administration of a compound that reduces myloid formation The selected subject may be a patient suffering from Parkinson's disease, Lewy body type. Dementia, incidental Lewy body disease, Lewy body variant of Alzheimer's disease, multiple system atrophy or pure autonomic failure, or any combination thereof. This may be the case.

[0022] In some embodiments, the inhibitor of amyloid formation is administered systemically or locally to the gut or central nervous system. It may be for administration as directed.

[0023] The present disclosure also provides a kit, comprising: a microbial Amyloid or microbial amyloid precursors and α-synuclein and / or other mammalian and amyloid or mammalian amyloid precursor, thereby enabling the methods of the present disclosure to be performed. In some embodiments, the microbial ami The microbial amyloid precursor contains CsgA.

[0024] The present disclosure provides a method of inhibiting amyloid formation in a subject in need thereof, comprising: The present invention relates to a compound described in the present specification, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. The method includes administering

[0025] The present disclosure provides a method for preventing or treating a disorder associated with amyloid formation in a subject in need thereof. A method of treating a disease comprising administering to a subject a compound described herein, or a pharmaceutically acceptable salt thereof, or provides a method comprising administering the pharmaceutical composition to a subject.

[0026] The present disclosure relates to a method of preventing or treating an amyloid disorder in a subject in need thereof. Thus, the compounds described herein, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof The method includes administering to a subject

[0027] In the methods of the disclosure, the amyloid disorder or disorder associated with amyloid formation is a neurological disorder. The disorder may be Parkinson's disease (PD), dementia with Lewy bodies, or multisystem atrophy, multiple sclerosis (MS), frontotemporal dementia (FTD), REM sleep behavior disorder (RB D), α-synucleinopathy, PD-related constipation, PD-related hypotension, Huntington's disease, It may be Alexander's disease, amyotrophic lateral sclerosis (ALS), or Alzheimer's disease. These disorders include intestinal dysbiosis, intestinal hyperpermeability, irritable bowel syndrome (IBS), and inflammatory bowel disease (IBD). The disease may be inflammatory bowel disease (IBD), ulcerative colitis or Crohn's disease.

[0028] The targets are dysphagia, decreased intestinal motility, gastroparesis, constipation (chronic constipation and chronic idiopathic constipation) (including constipation), small intestinal bacterial overgrowth (SIBO), diarrhea, abdominal pain and / or cramps, bloating , flatulence, hypersalivation (excessive salivation), anorectal dysfunction, bowel coordination disorder, and nausea The patient may suffer from gastrointestinal symptoms including one or more of the following: It may be associated with synucleinopathy, Parkinson's disease, or parkinsonism.

[0029] In certain embodiments, the amyloid disorder is characterized by the presence or level of enterobacterial amyloid aggregates. In some embodiments, the diagnosis can be made by detecting enterobacterial Diagnosing amyloid disorders by detecting the presence or level of amyloid protein In certain embodiments, the aggregates comprise bacterial CsgA protein. In some embodiments, the protein may include a bacterial CsgA protein. In certain embodiments, the disorder is characterized by the presence or levels of enterobacterial genes and gene transcripts. It can be diagnosed by detecting

[0030] The disclosed methods involve detecting bacterial proteins, such as CsgA, or their It may further comprise detecting the presence or level of a microorganism that produces the bacterial protein. In certain embodiments, the bacterial protein, the bacterial protein The presence of the microorganism producing the bacterial protein or the transcript mRNA of interest is detected. or in the intestinal sample, the bacterial protein or a bacterial protein-producing If the level of the microorganism occurring in the subject is greater than a predetermined level or a control, the subject is considered to have the prognosis. are selected as needing prevention or treatment.

[0031] The disclosed methods provide a method for treating amyloidosis, including the reduction or absence of intestinal amyloid aggregates following administration of a compound of the composition. The method may further include confirming or identifying the subject as exhibiting gastrointestinal symptoms. The disclosed methods include confirming a reduction or absence of intestinal amyloid protein after administration; It may further include identifying the subject as exhibiting a gastrointestinal symptom.

[0032] In some embodiments, the methods of the present disclosure comprise administering a level of enteric-coated amyloid. Some embodiments further include measuring or assessing the level and / or amyloid aggregation. In some embodiments, the disclosed methods may include administering a medicament containing amyloid-containing compound to a patient, the medicament being a medicament containing amyloid-containing compound, or a medicament containing amyloid-containing compound. The method further includes measuring or evaluating loid proteins.

[0033] The methods of the disclosure include detecting an amyloid disorder (e.g., Parkinson's disease, Levy disease) in a test subject. Dementia with Lewy bodies, incidental Lewy body disease, Lewy body variant of Alzheimer's disease, multisystem neurological disorders such as atrophy, or pure autonomic failure, or any combination thereof) or inhibiting the activity of a compound or composition described herein, in the presence or absence of the compound or composition. a plurality of concentrations of microbial amyloid or microbial amyloid precursor from the test subject; Multiple concentrations of alpha-synuclein and / or other mammalian proteins may be obtained from the biological sample obtained. contacting the animal amyloid or mammalian amyloid precursor; Analyzing or measuring the formation and comparing the analysis or measurement with a control analysis or measurement. wherein the control analyzes or measures amyloid formation in the absence of the composition. or a standard, e.g., a healthy subject or a subject with amyloidosis (e.g., Parkinson's disease, dementia with Lewy bodies, concomitant Lewy body disease, and Alzheimer's disease Bee body variant, multiple system atrophy, or pure autonomic failure, or any combination thereof and comparing the amount, rate, or formation of amyloid obtained from a subject suffering from and comparing the results with a control assay or measurement, which may include comparing the results with a control assay or measurement in the presence of the composition. amyloid formation is increased compared to amyloid formation in the absence of the composition. or the amount or rate or formation of amyloid in a sample from the test subject. However, amyloidosis may be induced by, for example, amyloidosis obtained from a control healthy subject or a control subject with amyloidosis. If the amount, rate or formation of amyloid is equal to or greater than the amount, rate or formation of amyloid, administering to said test subject an effective amount of a pharmaceutical composition suitable for inhibiting or treating said harm; In some further embodiments of these methods, the microorganisms are further comprised of: Biological amyloids or microbial amyloid precursors contain CsgA or essentially Csg A or CsgA.

[0034] In some embodiments, the methods described herein involve treating or inhibiting an amyloid disorder. identifying or selecting said subject as one who would benefit from the harm; Lewy body dementia, dementia with Lewy bodies, incidental Lewy body disease, Lewy body changes in Alzheimer's disease One or more of the following: atypical, multiple system atrophy, pure autonomic failure, or any combination of these Identifying or selecting such subjects as being at risk for or exhibiting symptoms of The method may further include: [Brief explanation of the drawings]

[0035] [Figure 1A] FIG. 1 is a graph showing crystal violet staining of wild-type E. coli biofilm growth after 4 days of static culture using the indicated concentrations of epigallocatechin gallate (EGCG), as assessed by optical density (OD). [Figure 1B] FIG. 11 is a graph showing in vitro αSyn aggregation measured by thioflavin T fluorescence during αSyn amyloid formation alone or in the presence of CsgA (25:1 molar ratio) with or without treatment with EGCG (50 μM). [Figure 2A-G]Figure 2A shows a series of graphs and images demonstrating that monocolonization with curli-sufficient bacteria induces increased αSyn-dependent pathology and inflammatory responses in the brain. Germ-free (GF) wild-type (WT) or Thy1-αSyn (ASO) animals were monocolonized with either wild-type, curli-sufficient E. coli (WT), or curli-deficient E. coli (ΔcsgBAC). Figure 2A shows total αSyn in whole brain lysates quantified by ELISA. Figure 2B shows quantification of insoluble αSyn fibrils in the striatum by dot blot assay. Figures 2C-2D show quantification of TNFα (Figure 2C) and IL-6 (Figure 2D) by ELISA from the striatum. Figures 2E-2G show the results of staining thin sections of brains from ASO mice. Sections were stained for Iba1 (microglia) to generate 3D cellular reconstructions and quantify the morphological characteristics of microglia present in the striatum. n = 3 (Figures 2A-2B), n = 6-7 (Figures 2C-2D), and n = 4 (Figures 2E-2G) (averaged from 20-40 cells for diameter or 5-7 cells for branching). Points represent individuals, and bars represent means and standard errors. Data analyzed by one-way ANOVA with Tukey's post-hoc test for Figures 2A-2D, or one-way ANOVA with two-tailed t-test for Figures 2E and 2F. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. [Figure 2H-J] Figure 2H shows levels of csgA in human fecal samples (Figure 2I), wild-type mice colonized with microbes from individuals with PD or matched controls (Figure 2I), or Thy1-αSyn (ASO) mice colonized with microbes from individuals with PD or matched controls (Figure 2J). Consistent with these data, csgA is predicted to be abundant in microbes from individuals with PD. [Figure 3A-I]Figure 3A-3G shows a series of graphs demonstrating that intestinal curli promotes progressive synuclein-dependent pathophysiology. Conventionally housed Thy1-αSyn (ASO) animals were intraintestinal injected with 30 μg of synthetic CsgA hexamer (CsgA; N-QYGGNN-C) or a non-amyloidogenic peptide (N122A; N-QYGGNA-C). For Figures 3A-3G, locomotor and GI function were examined over time at 0, 7, 21, and 70 days post-injection in beam traversal (Figure 3A), pole descent (Figure 3B), adhesive removal (Figure 3C), hindlimb clasping score (Figure 3D), wire hang (Figure 3E), and fecal excretion assessment (day 70) (Figure 3F). Figure 3G shows a principal component analysis of the compiled locomotor scores from Figures 3A-3F. Figures 3H-3I show quantification of insoluble αSyn fibrils in the striatum (Figure 3H) and ventral midbrain (Figure 3I) by dot blot assay. n = 8 (Figure 3A-3G), n = 4 (Figure 3H). Points represent individuals, and bars represent means and standard errors. Shown above individual time points are time courses analyzed by two-way ANOVA with Sidak post-hoc tests for between-group comparisons, and brackets indicate significance between treatments. Data from (Figure 3H) were analyzed by a two-tailed Mann-Whitney test. In Figures 3A-3I, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, and ****p ≤ 0.0001. [Figure 4A-B] 27A shows data from an in vivo experiment utilizing the procedure of Example 27A, demonstrating a reduction in csgA expression in E. coli MC4100 monocolonized mice treated with 30 mg / kg of compound 070 (A) or 30 mg / kg of compound 016 (B) on day 5 compared to mice treated with vehicle alone. [Figure 5] Figure 5 shows the number of fecal pellets excreted over a 15-minute interval when mice were 12-14 weeks of age. Figure 5 demonstrates an overall increase in the number of fecal pellets when mice were treated with compound 004a compared to the MC4100 control chow group (a qualitative trend was observed, but did not reach statistical significance). Points represent individual animals, and bars represent the mean and standard error. Data analyzed by one-way ANOVA with Dunnett's multiple comparison test. [Figure 6]Figure 6 shows a graph of fecal moisture content measured when animals were 12-14 weeks of age. Figure 6 demonstrates a significant decrease in percent moisture content in the MC4100 control chow group, suggesting a disease phenotype. Treatment with compound 004a reversed the curli operon-induced constipation phenotype. Points represent individual animals, and bars represent the mean and standard error. Data analyzed by one-way ANOVA with Dunnett's post-hoc test for between-group comparisons are shown above each treatment group, and brackets indicate significance between treatments; *p≦0.05, **p≦0.01. [Figure 7] Figure 7 shows the time it took to descend from the pole to the home cage at 18-20 weeks of age. Figure 7 demonstrates increased time in the MC-4100 control chow group (p=0.0741) compared to the KO CsgA group, suggesting a diseased motor phenotype. Compound 004a treatment showed a trend toward amelioration of the phenotype. Points represent individual animals, and bars represent the mean and standard error. Data analyzed by one-way ANOVA with Dunnett's post-hoc test. [Figure 8A-D] This is a series of graphs showing the reduction of csgA expression by compound 004a treatment at an early age of 12–14 weeks of age in mice monocolonized with E. coli MC4100. This effect was repeated and observed again at 14–16 weeks of age. Figure 8A shows csgA expression relative to the expression of the housekeeping gene recA at 12–14 weeks of age. Figure 8B shows csgA expression relative to the expression of the housekeeping gene cysG at 12–14 weeks of age. Figure 8C shows csgA expression relative to the expression of the housekeeping gene recA at 14–16 weeks of age. Figure 8D shows csgA expression relative to the expression of the housekeeping gene cysG at 14–16 weeks of age. Points represent individual animals, and bars represent the mean and standard error. Data analyzed by one-way ANOVA with Dunnett's post-hoc test for between-group comparisons are shown above each treatment group, and brackets indicate significance between treatments: *p ≤ 0.05; **p ≤ 0.01. DETAILED DESCRIPTION OF THE INVENTION

[0036] The majority of cases of neurodegenerative diseases are idiopathic, which has traditionally led to the widespread failure of most such diseases. Emerging theories have made it difficult to pinpoint the cause of most neurodegenerative disorders. The disease begins in the periphery rather than the brain or central nervous system (CNS) and progresses slowly over many years. However, the molecular mechanisms underlying the cellular mechanisms of cellular function in the periphery are unclear. The etiology of Parkinson's disease was the subject of research. In the case of Parkinson's disease, Constipation and hyposmia often occur in many patients decades before the onset of stereotyped motor symptoms. Therefore, without being limited by theory, it is known that α-synuclein Aggregation begins in the gastrointestinal (GI) tract and olfactory bulb, and aggregated α-synuclein propagates in a prion-like manner. It is thought that the process of the brain gradually progresses through the brain. More generally, Braak's hypothesis is In this known scenario, analysis of the molecular mechanisms involved in these peripheral tissues is crucial for Parkinson's disease. Preventing and / or treating amyloid disorders, such as alpha-synucleinopathies, including Johnson's disease This may lead to non-intuitive and non-traditional approaches to

[0037] Without being limited by theory, one molecular mechanism contemplated herein involves: Nucleate or otherwise lead to α-synuclein aggregation, thereby ultimately These disorders often result in Lewy body deposition and the clinical symptoms of Parkinson's disease and other α-synucleinopathies. Bacterial amyloid is involved as a seeding factor, initiating the pathological process. Bacterial proteins are aggregated forms of secreted bacterial proteins that mediate bacterial adhesion and proliferation to host cells. It is believed that they play a role in both the formation of iofilms and the establishment of the correct environment and aggregation. Without being limited by theory, in the presence of host proteins that are susceptible to bacterial amyloidosis, The nucleotide sequence itself acts as a direct structural template for host protein aggregation in a prion-like manner. Bacterial chaperone mechanisms involved in driving bacterial amyloid aggregation also uses host proteins as substrates, thereby promoting the production of amyloids from host proteins. Once aggregated, the aggregation of host proteins can persist for many years. These aggregates persist in a prion-like manner through the enteric nervous system. Finally, these aggregates This effect also affects the host tissues, resulting in the typical clinical symptoms of Parkinson's disease. Other amyloid diseases, such as Alzheimer's disease, which involve aggregation of proteins A-beta and / or tau Consistent with this, currently published human microbiomes Analysis of the Biome dataset revealed that E. coli-derived curli bacteria were more prevalent in Parkinson's disease patients. We found that the expression of the related csgA gene was increased, and we were able to identify the fecal microbiota from PD patients. Transplantation of GF into wild-type or ASO mice was performed using PICR of 16s rRNA sequences. Based on USt attribution analysis, greater csgA expression was observed in the microbiota compared with those from healthy controls. and d) provide abundances (e.g., d) of ... See WO 2019 / 028456 and the references cited therein. Intestinal amyloid aggregates are a potential cause of symptoms associated with Parkinson's disease and other amyloid disorders. These compounds inhibit and / or disrupt amyloid aggregation. Treating animals with α-amyloid-β-glucan improves these symptoms associated with Parkinson's disease and other amyloid disorders. It can be improved.

[0038] The present disclosure provides compounds, compositions, and methods for treating, ameliorating, or preventing amyloid disorders. The compounds disclosed herein are directed to the synthesis of the eukaryotic protein α-synuclein. Alters the ability of bacterial amyloid to promote aggregation and amyloid formation. The extent, rate of formation, stability, and / or rate of degradation of microbial-induced amyloid, or Any combination of alterations may be included. Compounds and compositions useful for treating or inhibiting neurodegenerative diseases and methods, and compounds, compositions and methods useful for preventing or ameliorating the progression of neurodegenerative diseases - Patents.com Further disclosed herein are compounds for treating or inhibiting gastrointestinal dysfunction associated with neurodegenerative diseases. Useful compounds, compositions and methods are further disclosed herein. The molecular pathogenesis of the disease and the molecular relationship between bacterial and mammalian amyloidogenesis Further disclosed herein are methods for studying associations. The compositions may comprise, consist essentially of, or consist of the compounds described herein. The present disclosure further provides an evaluation of the aggregation and disaggregation of amyloid proteins in both the host and bacteria. The methods of the present disclosure also relate to methods for facilitating the evaluation of drugs that affect these processes. It is also useful for identifying candidate compounds.

[0039] In some embodiments, a composition comprising or consisting essentially of a compound described herein. or a composition comprising the same, Seeding of α-synuclein aggregation by soluble amyloid or α-synuclein aggregation in vivo and are useful in preventing the formation of microbial amyloids that can disseminate disease, or in preventing such chemical reactions. The compounds and compositions are useful for preventing or treating Parkinson's disease and / or other alpha-synucleinopathies. (See, e.g., Example 26 and Tables 3 and 4.) ).

[0040] In some embodiments, a composition comprising or consisting essentially of a compound described herein. The composition comprising or containing α-amyloid, with or without seeding by microbial amyloid, -synuclein aggregation, thereby preventing microbial amyloid It is contemplated that these compounds may be useful in preventing or treating unrelated alpha-synucleinopathies. (See, for example, Example 26 and Tables 3 and 4).

[0041] In some embodiments, a compound comprising or derived essentially from a compound as described herein. The composition targets or consists of α-synuclein seeded by microbial amyloid. It may be useful to prevent nuclein aggregation, thereby preventing, for example, microbial amyloid. may have therapeutic benefit when administered to sites where ATP may be abundant (e.g., the gastrointestinal tract, etc.). It is intended that

[0042] In some embodiments, the composition comprises, consists essentially of, or is a compound described herein. or a composition comprising the same in Parkinson's disease and other alpha-synucleinopathies It is contemplated that this may have therapeutic benefits. The advantage of these compounds is that they inhibit the aggregation of α-synuclein and / or microbial amyloid. For compounds that inhibit more than one type of aggregation, this may be due to inhibition. It is further contemplated that these inhibitory effects may be additive or synergistic (see, e.g., Example 2). 6 and Table 2).

[0043] Thus, an amyloid disorder, e.g., any of the amyloid disorders in Table 2 (below), e.g., For example, α-synucleinopathy, Parkinson's disease, dementia with Lewy bodies, incident Lewy body dementia, Somatoform disorders, Lewy body variant of Alzheimer's disease, multiple system atrophy, pure autonomic failure, or inhibits, ameliorates, reduces the likelihood, or delays the onset of any combination of two or more of the listed items. Provided herein are compounds useful for the prolongation, treatment and / or prevention of rheumatoid arthritis.

[0044] Compounds of the Invention In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof: [ka] During the ceremony, A 1 is -C(R 7 )(R 8 ) or -CO-, or X is -SO2-, CO or Kuha-C(R 9 )(R 10 ), then A 1 -N(R 7 )- and A 2 is not present or -C(R 5 )(R 6 ) and L 1 is a bond, (-CH2-) m , -CF2-, -(C=O)- or -C(R 9 )(R 10 )- and L 2 is a bond, (-CH2-) m , -CF2-, -(C=O)- or -C(R 9 )(R 10 )- and X is -N(R 11 )-, -N(R 14 )-, -O-, -CO-, -S-, -S(=O )-, -SO2-, -CF2-, -C(R 9 )(R 10 )- and Y is O or S; Z is =C(R 13 )-, =N-, or -N(R 11 )- and R 1 is substituted or unsubstituted phenyl, substituted or unsubstituted heterocyclyl, R 2 is a substituted or unsubstituted naphthyl, a substituted or unsubstituted phenyl, a substituted or unsubstituted Substituted quinolinyl, substituted or unsubstituted isoquinolinyl, or substituted or unsubstituted heteroquinolinyl It is a krill, R 3 and R 4 is independently absent or -H, substituted or -, where valence permits. Unsubstituted C1~C 10 Alkyl, acyl, -CO2R 7 , -CON(R 7 )(R 8 ), -P ═O(OH) or —SO(OH), R 5 and R 6 is independently absent or -H and C1 to C2 as far as valence allows. 10 a or in combination with spirocarbocyclic or spiro(hetero) forming a carbocyclic ring, R 7 and R 8 is independently absent or -H, substituted or -, where valence permits. Unsubstituted C1~C 10 Alkyl, -(CH2) m -aryl, -(CH2) m -Hetero Ants -(CH2) m-substituted or unsubstituted cycloalkyl, -R 14 Is it selected from or combined to form a spiropentanyl ring, R 9 and R 10 independently, at each occurrence, -H, -Cl, -Br, -F, -CF3 , C1~C 10 is alkyl, R 11 and R 12 is independently, at each occurrence, -H, acyl, sulfonyl, substituted or unsubstituted. <Unsubstituted C1-C 10 Alkyl, C3-C6 cycloalkyl, C 3~6 Heterocyclyl , substituted or unsubstituted benzyl, -(CH2) o -(substituted or unsubstituted aryl) or -(CH2) o -(substituted or unsubstituted heteroaryl), R 13 -H, -OH, -OR 11 , -Cl, -Br, -F, -CN, -CF3, - CH2F, -CHF2, substituted or unsubstituted C1-C 10 Alkyl, C1-C 10 Alkenyl , C3-C6 cycloalkyl, substituted or unsubstituted C3-C6 heterocycloalkyl, acyl , -CO2R 7 , -(CH2) m CO2N(R 11 )(R 12 ), -CON(R 7 )(R 8 ), -(CH2) m OH, -(CH2) m CO2H, -(CH2) m NH2, -(CH 2) m N(R 11 )(R 12 ), -N(R 11 )(R 12 ), -NR 11 (C=O)(C H2) m CH3, -NR 11 (C=O)R 12 and NR 12 (SO2)(CH2) m CH Selected from 3, R 14 -H, C1~C 10 Alkyl, C1-C 10 Alkenyl, C1-C 10 (Mo (mono or poly)hydroxylated alkyl, -(CH2) o -R 15 , -(CH2CH2O) o -R 15 , -(CH2) m -CO2H, -(CH2) m -NH2, -(CH2) m -( CO)NR 16 R 17 or a protecting group, R 15 is -CON(R 11 )(R 12) , -N(R 11 )(R 12 ), acyl, -C O2R 7 , substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cyclohexyl cycloalkyl, substituted or unsubstituted heterocycloalkyl; R 16 and R 17 are independently -H or -CH3; m is independently 0 to 10 in each occurrence; n is, independently in each occurrence, 1 to 5; o is independently 1 to 20 in each occurrence; [ka] represents a single bond or a double bond, However, the compound of formula (I) [ka] isn't it.

[0045] In some embodiments, the compound of formula (I) is formula (Ia) or a pharmaceutically acceptable salt thereof: A salt comprising: [ka] During the ceremony, A 1 is -C(R 7 )(R 8 ) or -CO-, or X is -SO2-, CO or Kuha-C(R 9 )(R 10 ), then A 1 -N(R 14 )- and A 2 is not present or -C(R 5 )(R 6 ) and L 1 is a bond, (-CH2-) m , -CF2-, -(C=O)- or -C(R 9 )(R 10 )- and L 2 is a bond, (-CH2-) m , -CF2-, -(C=O)- or -C(R 9 )(R 10 )- and X is -N(R 11 )-, -O-, -CO-, -S-, -S(=O)-, -SO2-, -CF2-, -C(R 9 )(R 10 )- and Y is O or S; Z is =C(R 13 )-, =N-, or -N(R 11 )- and R1 is substituted or unsubstituted phenyl, substituted or unsubstituted heterocyclyl, R 2 is a substituted or unsubstituted naphthyl or a substituted or unsubstituted heterocyclyl; R 3 and R 4 is independently absent or -H, substituted or -, where valence permits. Unsubstituted C1~C 10 Alkyl, acyl, -CO2R 7 , -CON(R 7 )(R 8 ), -P ═O(OH) or —SO(OH), R 5 and R 6 is independently absent or -H and C1 to C2 as far as valence allows. 10 a or in combination with spirocarbocyclic or spiro(hetero) forming a carbocyclic ring, R 7 and R 8 is independently absent or -H, substituted or -, where valence permits. Unsubstituted C1~C 10 Alkyl, -(CH2) m -aryl, -(CH2) m -Hetero Ants -(CH2) m -substituted or unsubstituted cycloalkyl, -R 14 Is it selected from or combined to form a spiropentanyl ring, R 9 and R 10 independently, at each occurrence, -H, -Cl, -Br, -F, -CF3 , C1~C 10 is alkyl, R 11 and R 12 is independently, at each occurrence, -H, acyl, sulfonyl, substituted or unsubstituted. <Unsubstituted C1-C 10Alkyl, C3-C6 cycloalkyl, substituted or unsubstituted benzene Zill, -(CH2) o -(substituted or unsubstituted aryl) or -(CH2) o -(substitute young or unsubstituted heteroaryl), R 13 -H, -OH, -OR 11 , -Cl, -Br, -F, -CN, -CF3, - CH2F, -CHF2, substituted or unsubstituted C1-C 10 Alkyl, C1-C 10 Alkenyl , C3-C6 cycloalkyl, substituted or unsubstituted C3-C6 heterocycloalkyl, acyl , -CO2R 7 , -(CH2) m CO2N(R 11 )(R 12 ), -CON(R 7 )(R 8 ), -(CH2) m OH, -(CH2) m CO2H, -(CH2) m NH2, -(CH 2) m N(R 11 )(R 12 ), -N(R 11 )(R 12 ), -NR 11 (C=O)(C H2) m CH3, -NR 11 (C=O)R 12 and NR 12 (SO2)(CH2) m CH Selected from 3, R 14 -H, C1~C 10 Alkyl, C1-C 10 Alkenyl, C1-C 10 (Mo (mono or poly)hydroxylated alkyl, -(CH2) o -R 15 , -(CH2CH2O) o-R 15 , -(CH2) m -CO2H, -(CH2) m -NH2, -(CH2) m -( CO)NR 16 R 17 or a protecting group, R 15 is -CON(R 11 )(R 12) , -N(R 11 )(R 12 ), acyl, -C O2R 7 , substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cyclohexyl cycloalkyl, substituted or unsubstituted heterocycloalkyl; R 16 and R 17 are independently -H or -CH3; m is independently 0 to 10 in each occurrence; n is, independently in each occurrence, 1 to 5; o is independently 1 to 20 in each occurrence; [ka] represents a single bond or a double bond, provided that the compound of formula (Ia) is or [ka] isn't it.

[0046] In some embodiments, X is -SO2-. In some embodiments, X is -NR 14 In some embodiments, X is -S-.

[0047] In some embodiments, A 1 is -C(R 7 )(R 8). In some embodiments So, A 1 is -CO-. In some embodiments, A 1 X is -SO2-, C O or -C(R 9 )(R 10 ), then -N(R 7 )- or -N(R 14 )-is In certain embodiments, A 1 is -N(R 7 )- or -N(R 14 )- and X is , -SO2-. In certain embodiments, A 1 is -N(R 7 )- and X is In certain embodiments, A 1 is -N(R 14 )- and X is -SO2-.

[0048] In some embodiments, A 2 is absent. In some embodiments, A 2 -C (R 5 )(R 6 )-.

[0049] In some embodiments, L 1 is -(CH2)- m In some embodiments, , L 1 is —(CH)—. In some embodiments, L 1 is -CF2- In some embodiments, L 1 is —(C═O)—. In some embodiments, L 1 is -C(R 9 )(R 10 In some embodiments, L 1 is a bond do.

[0050] In some embodiments, L 2 is -(CH2)- m In some embodiments, , L 2 is —(CH)—. In some embodiments, L 2 is -CF2- In some embodiments, L 2 is —(C═O)—. In some embodiments, L 2 is -C(R 9 )(R 10 In some embodiments, L 2 is a bond do.

[0051] In some embodiments, X is —N(R 11 In some embodiments, X is -O-. In some embodiments, X is -CO-. In some embodiments, X is -S-. In some embodiments, X is -S(=O)-. In some embodiments, X is -SO2-. In some embodiments, X is In some embodiments, X is C(R 9 )(R 10 )-. In some embodiments, X is —N(R 14 )-.

[0052] In some embodiments, Y is O. In some embodiments, Y is S.

[0053] In some embodiments, Z is ═C(R 13 In some embodiments, Z is =N- or -N(R 11 In some embodiments, Z is —N(R 1 1 )-.

[0054] In some embodiments, R 1 is unsubstituted phenyl. In some embodiments, R 1 is substituted phenyl. In some particular embodiments, R 1 is trifluoromethyl In some embodiments, R 1 is unsubstituted heterocyclyl. In some embodiments, R 1 is a substituted heterocyclyl.

[0055] In some embodiments, R 2 is unsubstituted naphthyl (e.g., 1-naphthyl or 2-naphthyl). In some embodiments, R 2 is a substituted naphthyl (e.g., 1-naphthyl) In some embodiments, R 2 is an unsubstituted heterocyclyl In some embodiments, R 2 is a substituted heterocyclyl. In terms of form, R 2 is unsubstituted phenyl. In some embodiments, R 2 is a substitution function. phenyl (e.g., tert-butyl-substituted phenyl, or biphenyl). In the embodiment, R 2 is unsubstituted quinolinyl (e.g., 4-quinolinyl, 5-quinolinyl or 8-quinolinyl). In some embodiments, R 2 is a substituted quinolinyl (e.g., In some embodiments, the quinolinyl group is 4-quinolinyl, 5-quinolinyl, or 8-quinolinyl. , R 2 is unsubstituted isoquinolinyl (e.g., 4-isoquinolinyl, 5-isoquinolinyl or is 8-isoquinolinyl). In some embodiments, R 2 is a substituted isoquinolinyl (for example, 4-isoquinolinyl, 5-isoquinolinyl or 8-isoquinolinyl).

[0056] In some embodiments, R 3 is —H. In some embodiments, R 3 is non Substitution C1~C 10 In some embodiments, R 3 is the substitution C1~C 10 In some embodiments, R 3 is acyl. So, R 3 -CO2R 7 (e.g., —COH). In some embodiments, R 3 is -CON(R 7 )(R 8 In some embodiments, R 3 is -P=O (OH). In some embodiments, R 3 is -SO2(OH). In some embodiments, R 3 is -CO2H.

[0057] In some embodiments, R 4 is absent. In some embodiments, R 4 -H In some embodiments, R 4 is unsubstituted C1 to C 10 It is an alkyl. In some embodiments, R 4 is the substitution C1~C 10 In some embodiments, the aryl group is alkyl. , R 4 is acyl. In some embodiments, R 4 -CO2R 7 (for example,- COH). In some embodiments, R 4 is -CON(R 7 )(R8 ) In some embodiments, R 4 is -P=O(OH). In some embodiments, is R 4 is -SO2(OH).

[0058] In some embodiments, R 3 is -H, and R 4 is a substituted or unsubstituted C1-C 10 Alkyl, acyl, -CO2R 7 , -CON(R 7 )(R 8 ), -P=O(OH)2 or In some particular embodiments, R 3 is -H , R 4 -CO2R 7 (e.g., -CO2H).

[0059] In some embodiments, R 4 is -H, and R 3 is a substituted or unsubstituted C1-C 10 Alkyl, acyl, -CO2R 7 , -CON(R 7 )(R 8 ), -P=O(OH)2 or In some particular embodiments, R 4 is -H , R 3 -CO2R 7 (e.g., -CO2H).

[0060] In some particular embodiments, R 4 is -Me and R 3 -CO2R 7 (example For example, -CO2H).

[0061] In some embodiments, R 5 and R 6is A 2 If it doesn't exist, it doesn't exist. In some embodiments, R 5 and R 6 are both -H. In some embodiments, R 5 and R 6 Both are C1~C 10 In some embodiments, R 5 exists Not present, R 6 -H and C1 to C 10 In some embodiments, the alkyl is selected from the group consisting of aryl, ... is R 5 and R 6 are combined to form a spirocarbocyclic or spiro(hetero)carbocyclic ring Form.

[0062] In some embodiments, R 7 is absent. In some embodiments, R 7 teeth,- H, substituted or unsubstituted C1-C 10 Alkyl, -(CH2) m -aryl, -(CH2) m -heteroaryl, -(CH2) m -substituted or unsubstituted cycloalkyl, -R 14 Choose from In some embodiments, R 7 is a substituted or unsubstituted C1-C 10 is alkyl In some embodiments, R 7 is -(CH2) m -substituted or unsubstituted cycloalkyl In some embodiments, R 7 is -(CH2) m -aryl. In embodiments, R 7 is —H. In some embodiments, R 7 is substituted or unsubstituted C 1~5In some embodiments, R 7 is -Me. Some In embodiments, R 7 is a substituted or unsubstituted C alkyl.

[0063] In some embodiments, R 8 is absent. In some embodiments, R 8 -H , substituted or unsubstituted C1 to C 10 Alkyl, -(CH2) m -aryl, -(CH2) m - Heteroaryl, -(CH2) m -substituted or unsubstituted cycloalkyl, -R 14 Selected from can be.

[0064] In some embodiments, R 7 and R 8 combine to form a spiropentanyl ring do.

[0065] In some embodiments, R 9 -H, -Cl, -Br, -F, -CF3 and C1~ C 10 In some embodiments, R 9 is selected from -H and -F It is selected.

[0066] In some embodiments, R 10 -H, -Cl, -Br, -F, -CF3 and C1 ~C 10 In some embodiments, R 10 -H and -F are selected.

[0067] In some embodiments, R 11 is -H, acyl, sulfonyl, substituted or unsubstituted C1 ~C 10Alkyl, C3-C6 cycloalkyl, and C 3~6 heterocyclyl can be.

[0068] In some embodiments, R 12 is -H, acyl, sulfonyl, substituted or unsubstituted C1 ~C 10 Alkyl, C3-C6 cycloalkyl and C 3~6 heterocyclyl do.

[0069] In some embodiments, R 13 is —H. In some embodiments, R 13 teeth , substitution C1~C 10 Alkyl (e.g., -(CH2) m CO2N(R 11 )(R 12 ), -CON(R 7 )(R 8 ), -(CH2) m OH, -(CH2) m CO2H, -(CH2 ) m NH2 or -(CH2) m N(R 11 )(R 12 )). Some embodiments So, R 13 is unsubstituted C1 to C 10 In some embodiments, R 13 is -N(R 11 )(R 12 In some embodiments, R 13 is -NR 11 (C=O)(CH2) m CH3, -NR 11 (C=O)R 12 or NR 12 (SO2)( CH2) m In some embodiments, R1 is CH3. 3 is -Br or -Cl .

[0070] In some embodiments, R 14 is —H. In some embodiments, R 14 teeth , C1~C 10 Alkyl or C1-C 10 In some embodiments, R 14 is C1~C 10 Some examples are (mono- or poly)hydroxylated alkyls. In the embodiment, R 14 is -(CH2) o -R 15 In some embodiments, R 1 4 is -(CH2CH2O) o -R 15 In some embodiments, R 14 teeth,- (CH2) m In some embodiments, R 14 is -(CH2) m - NH2. In some embodiments, R 14 is -(CH2) m -(CO)NR 16 R 17 In some embodiments, R 14 is a protecting group.

[0071] In some embodiments, R 15 is -CON(R 11 )(R 12 ) are some In an embodiment of the present invention, R 15 is -N(R 11 )(R 12 ). In some embodiments, , R 15 is acyl. In some embodiments, R 15 -CO2R 7 is. In some embodiments, R 15 is substituted aryl. In some embodiments, R 15 is unsubstituted aryl. In some embodiments, R 15 is a substituted heteroaryl In some embodiments, R 15 is an unsubstituted heteroaryl In some embodiments, R 15 is substituted cycloalkyl. So, R 15 is unsubstituted cycloalkyl. In some embodiments, R 15 is placed In some embodiments, R 15 is an unsubstituted heterocyclic It is a hydroxyalkyl.

[0072] In some embodiments, R 16 is —H. In some embodiments, R 16 teeth , -CH3.

[0073] In some embodiments, R 17 is —H. In some embodiments, R 17 teeth , -CH3.

[0074] In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, m is 3. In some embodiments, m is 4. In some embodiments, m is 5. In embodiments, m is 6. In some embodiments, m is 7. In some embodiments, m is 8. In some embodiments, m is 9. So m is 10.

[0075] In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

[0076] In some embodiments, o is 1. In some embodiments, o is 2. In some embodiments, o is 3. In some embodiments, o is 4. In some embodiments, o is 5. In some embodiments, o is 6. In embodiments, o is 7. In some embodiments, o is 8. In some embodiments, o is 9. In some embodiments, o is 10. In some embodiments, o is 11. In some embodiments, o is 12. In some embodiments, o is 13. In some embodiments, o is 14. In some embodiments, o is 15. In some embodiments, o is 16. In some embodiments, o is 17. In some embodiments, o is 18. In some embodiments, o is 19. In some embodiments, o is 20.

[0077] In any of the preceding embodiments, formula (I) may be selected from the group consisting of formula (II), ( III), (IV), (V), (VI), (VII), (VIII), (IX), and ( X).

[0078] In some embodiments, Formula (I) has the structure of Formula (II): [ka]

[0079] In some embodiments, Formula (I) has the structure of Formula (III): [ka]

[0080] In some embodiments, Formula (I) has the structure of Formula (IV): [ka]

[0081] In some embodiments, Formula (I) has the structure of Formula (V): [ka]

[0082] In some embodiments, Formula (I) has the structure of Formula (VI): [ka]

[0083] In some embodiments, Formula (I) has the structure of Formula (VII): [ka]

[0084] In some embodiments, Formula (I) has the structure of Formula (VIII): [ka]

[0085] In some embodiments, Formula (I) has the structure of Formula (IX): [ka]

[0086] In some embodiments, Formula (I) has a structure of Formula (X): [ka]

[0087] In some embodiments, Formula (II) has the structure of Formula (IIa) or Formula (IIb): : [ka] During the ceremony, L 1 is a bond, (-CH2-) m , —CF—, or —(C═O)—; L 2 is a bond, (-CH2-) m , —CF—, or —(C═O)—; m is independently 0 to 10 in each occurrence; R 1 is substituted or unsubstituted phenyl, substituted or unsubstituted heterocyclyl, R 2 is a substituted or unsubstituted naphthyl or a substituted or unsubstituted heterocyclyl , R 3 is -CO2H, [ka] and R 5 and R 6 are H, respectively, R 7 is a substituted or unsubstituted C1 to C 10 Alkyl, or substituted or unsubstituted cycloalkenyl It is Lukil, R 13 are -H, -Cl, -Br, -F, -CN, -CF3, -CH2F, -CHF2 , substituted or unsubstituted C1 to C 10 Alkyl, -NH2, -CONH2, -(CH2)-N( CH3)2, -NH(cyclopentyl), -NH(benzyl), -NH(tetrahydropyridine) oran), -NH-(CH2)(cyclopentyl) and -O-(CH2)2-phenyl be selected.

[0088] In some embodiments, Formula (II) has the structure of Formula (IIc): [ka] During the ceremony, L 2 is a bond, (-CH2-) m , —CF—, or —(C═O)—; m is independently 0 to 10 in each occurrence; R 2 is a substituted or unsubstituted naphthyl or a substituted or unsubstituted heterocyclyl , R 3 is -CO2H, [ka] and R 5 and R 6 are H, respectively, R 7 is a substituted or unsubstituted C1 to C 10 Alkyl, or substituted or unsubstituted cycloalkenyl It is Lukil, R 13 are -H, -Cl, -Br, -F, -CN, -CF3, -CH2F, -CHF2 , substituted or unsubstituted C1 to C 10Alkyl, -NH2, -CONH2, -(CH2)-N( CH3)2, -NH(cyclopentyl), -NH(benzyl), -NH(tetrahydropyridine) oran), -NH-(CH2)(cyclopentyl) and -O-(CH2)2-phenyl be selected.

[0089] In some embodiments, formula (IX) has the structure of formula (IXa) or formula (IXb): : [ka] During the ceremony, L 1 is a bond, (-CH2-) m , —CF—, or —(C═O)—; L 2 is a bond, (-CH2-) m , —CF—, or —(C═O)—; m is independently 0 to 10 in each occurrence; o is independently 1 to 20 in each occurrence; R 1 is substituted or unsubstituted phenyl, substituted or unsubstituted heterocyclyl, R 2 represents substituted or unsubstituted naphthyl, substituted or unsubstituted quinolinyl, substituted or unsubstituted isoquinolinyl, substituted phenyl, or substituted or unsubstituted heterocyclyl; R 3 is -CO2H, [ka] and R 13 are -H, -Cl, -Br, -F, -CN, -CF3, -CH2F, -CHF2 , -(CH2) m -NMe2, -CONH2, -CO2H, and substituted or unsubstituted C1-C 10 alkyl, R 14 is C1~C 10 Alkyl, -(CH2) o -(unsubstituted cycloalkyl), -( CH2) o -(substituted or unsubstituted phenyl), -(CH2) o -naphthyl or -(CH2) o -biaryl.

[0090] In some embodiments, Formula (IX) has the structure of Formula (IXc): [ka] During the ceremony, L 2 is a bond, (-CH2-) m , —CF—, or —(C═O)—; m is independently 0 to 10 in each occurrence; o is independently 1 to 20 in each occurrence; R 2 represents substituted or unsubstituted naphthyl, substituted or unsubstituted quinolinyl, substituted or unsubstituted isoquinolinyl or substituted or unsubstituted heterocyclyl; R 3 is -CO2H, [ka] and R 13 are -H, -Cl, -Br, -F, -CN, -CF3, -CH2F, -CHF2 , -(CH2) m -NMe2, -CONH2, -CO2H, and substituted or unsubstituted C1-C 10 alkyl, R 14 is C1~C 10Alkyl, -(CH2) o -(unsubstituted cycloalkyl), -( CH2) o -(substituted or unsubstituted phenyl), -(CH2) o -naphthyl or -(CH2) o -biaryl.

[0091] In certain embodiments, the compound of formula (II) is selected from the group consisting of Compound 015a, Compound 016b, Compound 016c, Compound 016d, Compound 016e, Compound 016f, Compound 016h, Compound 016i, Compound 016j ... Compound 024, Compound 042, Compound 052, Compound 068, Compound 070, Compound 071, Compound 072, Compound 074, Compound 108, Compound 109, Compound 116, Compound 12 0, compound 121, compound 122, compound 123, compound 126, compound 127, compound 128, Compound 138, Compound 172, Compound 175, Compound 176, Compound 177, Chemical Compound 183, and Compound 184.

[0092] In certain embodiments, the compound of formula (V) is selected from the group consisting of Compound 051, Compound 057, and Compound 0 62 are selected.

[0093] In certain embodiments, the compound of formula (VII) is selected from compound 022 and compound 053. It is selected.

[0094] In certain embodiments, the compound of formula (IX) is selected from the group consisting of Compound 003, Compound 004, Compound 005, Compound 006, Compound 007, Compound 008, Compound 009, Compound 010, Compound 110, Compound 111, Compound 112, Compound 113, Compound 114, Compound 115, Compound 116, Compound 117, Compound 118, Compound 119, Compound 120, Compound 121, Compound 122, Compound 123, Compound 124, Compound 125, Compound 126, Compound 127, Compound 128, Compound 129, Compound 130, Compound 131, Compound 132, Compound 133, Compound 134, Compound 135, Compound 136, Compound 137, Compound 138, Compound 139, Compound 140, Compound 1 Compound 004a, Compound 004b, Compound 140, Compound 161, Compound 162, Compound 16 3, compound 164, compound 167, compound 178, compound 179, compound 180, compound 181, compound 198, compound 200, compound 202, compound 207, compound 208, or is selected from Compound 216.

[0095] In certain embodiments, the compound of formula (X) is selected from compound 188 and compound 193. can be.

[0096] In certain embodiments, the compounds of formula (I) are selected from the group consisting of the compounds of Table 1 and pharmaceutically acceptable salts thereof. salts. [Table 1] TIFF2026016368000027.tif204170JPEG2026016368000028.jpg212170TIFF2026016368000029.tif204170TIFF2026016368000030.tif246170TIFF2026016368000031.tif246170TIFF2026016368000032.tif246170TIFF2026016368000033.tif194170TIFF2026016368000034.tif238170TIFF2026016368000035.tif221170TIFF2026016368000036.tif246170TIFF2026016368000037.tif229170TIFF2026016368000038.tif254170TIFF2026016368000039.tif254170TIFF2026016368000040.tif246170TIFF2026016368000041.tif229170TIFF2026016368000042.tif221170TIFF2026016368000043.tif229170TIFF2026016368000044.tif238170TIFF2026016368000045.tif246170TIFF2026016368000046.tif204170TIFF2026016368000047.tif254170TIFF2026016368000048.tif246170TIFF2026016368000049.tif229170TIFF2026016368000050.tif229170TIFF2026016368000051.tif229170TIFF2026016368000052.tif254170TIFF2026016368000053.tif254170TIFF2026016368000054.tif254170TIFF2026016368000055.tif254170TIFF2026016368000056.tif254170TIFF2026016368000057.tif111170

[0097] In certain embodiments, the compound of formula (I) is Compound 001, Compound 003, Compound 00 4, Compound 004a, Compound 004b, Compound 070, Compound 094, Compound 108, Chemical Compound 109, Compound 116, Compound 122, Compound 140, and pharmaceutically acceptable salts thereof salts.

[0098] The above compounds and pharmaceutically acceptable salts thereof are collectively referred to as the compounds of the present invention. can.

[0099] The following compounds are referenced herein: reference compound [ka]

[0100] The term "alkyl" refers to a straight or branched chain saturated hydrocarbon having from 1 to 20 carbon atoms. Elementary radical (``C 1~20 In some embodiments, alkyl refers to a radical of The group has 1 to 10 carbon atoms ("C 1~10 alkyl"). In some embodiments In the example, the alkyl group has 1 to 9 carbon atoms ("C 1~9 alkyl"). In this embodiment, the alkyl group has 1 to 8 carbon atoms ("C 1~8 alkyl In some embodiments, the alkyl group has 1 to 7 carbon atoms ("C 1~7 a In some embodiments, the alkyl group has 1 to 6 carbon atoms ("alkyl"). C 1~6 In some embodiments, an alkyl group contains 1 to 5 carbon atoms. ("C 1~5 In some embodiments, the alkyl group is selected from 1 to 4 alkyl groups. carbon atoms ("C 1~4 In some embodiments, the alkyl group is having 1 to 3 carbon atoms ("C 1~3 In some embodiments, alkyl The alkyl group has 1 to 2 carbon atoms ("C 1~2 alkyl"). In some embodiments In some embodiments, the alkyl group has one carbon atom (a "C alkyl"). In some embodiments, the alkyl group has 2 to 6 carbon atoms ("C 2~6 alkyl). C 1~ 6 Examples of alkyl groups include methyl (C1), ethyl (C2), and propyl (C3) (e.g., butyl (C4) (e.g., n-butyl, tert- butyl, sec-butyl, iso-butyl), pentyl (C5) (e.g., n-pentyl , 3-pentanyl, amyl, neopentyl, 3-methyl-2-butanyl, tertiary amyl) and hexyl (C6) (e.g., n-hexyl). Further examples of alkyl groups include: Examples of the alkyl groups include n-heptyl (C7) and n-octyl (C8). Unless otherwise specified, each instance of an alkyl group is independently unsubstituted (an "unsubstituted alkyl") or or substituted with one or more substituents (e.g., halogens such as F) ("substituted alkyl" In certain embodiments, the alkyl group is an unsubstituted C 1~10 Alkyl (e.g., unsubstituted exchange C 1~6 Alkyl, for example, -CH3 (Me), unsubstituted ethyl (Et), unsubstituted propyl propyl (Pr, e.g., unsubstituted n-propyl (n-Pr), unsubstituted isopropyl (i-Pr) ), unsubstituted butyl (Bu, for example, unsubstituted n-butyl (n-Bu), unsubstituted tert-butyl butyl (tert-Bu or t-Bu), unsubstituted sec-butyl (sec-Bu or sB u), unsubstituted isobutyl (i-Bu). In certain embodiments, the alkyl group is , replace C 1~10 Alkyl (e.g., substituted C 1~6 Alkyl, e.g., -CHF, -CHF 2, -CF3 or benzyl (Bn). The alkyl group may be branched or unbranched. may be.

[0101] The term "alkenyl" refers to an alkyl group having 1 to 20 carbon atoms and one or more carbon-carbon double bonds ( a radical of a straight-chain or branched hydrocarbon group having, for example, 1, 2, 3 or 4 double bonds In some embodiments, the alkenyl group has 1 to 20 carbon atoms ( "C 1~20 In some embodiments, the alkenyl group has 1 to 12 amino groups. having carbon atoms ("C 1~12 In some embodiments, alkenyl The alkyl group has 1 to 11 carbon atoms ("C 1~11 Some alkenyl In embodiments, the alkenyl group has 1 to 10 carbon atoms ("C 1~10 Arkeny In some embodiments, an alkenyl group has 1 to 9 carbon atoms ("C 1~9 In some embodiments, an alkenyl group has 1 to 8 carbon atoms. ("C 1~8 In some embodiments, the alkenyl group is selected from the group consisting of 1 to 10 alkyl groups. having seven carbon atoms ("C 1~7 In some embodiments, an alkene is an alkenyl. The aryl group has 1 to 6 carbon atoms ("C 1~6 alkenyl). In certain embodiments, the alkenyl group has 1 to 5 carbon atoms ("C 1~5 alkenyl). In some embodiments, the alkenyl group has 1 to 4 carbon atoms ("C 1~4 Al In some embodiments, the alkenyl group has 1 to 3 carbon atoms ( "C 1~3 In some embodiments, the alkenyl group is an alkyl group having 1 to 2 carbon atoms. atoms ("C 1~2 In some embodiments, the alkenyl group is having one carbon atom ("C1 alkenyl"); one or more carbon-carbon double bonds , internal (e.g., 2-butenyl) or terminal (e.g., 1-butenyl). 1~ 4 Examples of alkenyl groups include methylidenyl (C1), ethenyl (C2), 1-propenyl 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4) and the like. 1~6 Examples of alkenyl groups include the above-mentioned C 2~4 Alkenyl groups, as well as pentenyl (C5), pentadienyl (C5), and hexenyl Further examples of alkenyl include heptenyl (C7), octenyl (C8), and octyl (C9). Examples include ctenyl (C8), octatrienyl (C8), etc. Unless otherwise specified , each instance of an alkenyl group is independently unsubstituted (an "unsubstituted alkenyl"), or Substituted with one or more substituents ("substituted alkenyl"). In certain embodiments, alkenyl The phenyl group is an unsubstituted C 1~20 In certain embodiments, the alkenyl The group is a substituted C 1~20 In the alkenyl group, the stereochemistry is specified. No C=C double bond (e.g., -CH=CHCH3 or [ka] can be in the (E) or (Z) configuration.

[0102] The term "cycloalkyl" refers to a cyclic alkyl group having from 3 to 10 ring carbon atoms. Digital ("C 3~10 In some embodiments, cycloalkyl refers to The alkyl group has 3 to 8 ring carbon atoms ("C 3~8 Some In embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms ("C 3~6 Cyclo In some embodiments, the cycloalkyl group has 5 to 6 ring carbon atoms. ("C 5~6 In some embodiments, the cycloalkyl group is , having 5 to 10 ring carbon atoms ("C 5~10 Cycloalkyl). C 5~6 Cyclo Examples of alkyl groups include cyclopentyl (C5) and cyclohexyl (C5). C 3~6 Examples of cycloalkyl groups include the above-mentioned C 5~6 Cycloalkyl groups and Examples include cyclopropyl (C3) and cyclobutyl (C4). 3~8 Cycloalkane Examples of the alkyl group include the above-mentioned C 3~6 Cycloalkyl groups, as well as cycloheptyl (C7) and and cyclooctyl (C8). Unless otherwise specified, each of the cycloalkyl groups Examples are independently unsubstituted ("unsubstituted cycloalkyl") or one or more substituted In certain embodiments, the cycloalkyl is substituted with a group ("substituted cycloalkyl"). The group is an unsubstituted C 3~10 In certain embodiments, cycloalkyl is The alkyl group is a substituted C 3~10 It is cycloalkyl.

[0103] The term "acyl" refers to a group having the general formula -C(=O)R X1 , -C(=O)OR X1 , -C( =O)-OC(=O)R X1 , -C(=O)SR X1 , -C(=O)N(R X1 )2, -C(=S)R X1 , -C(=S)N(R X1 )2, and -C(=S)S(R X1 ), - C(=NR X1 )R X1 , -C(=NR X1 ) OR X1 , -C(=NR X1 )SR X1 , and -C(=NR X1 )N(R X1 )2, wherein R X1 is hydrogen, halo gen, substituted or unsubstituted hydroxyl, substituted or unsubstituted thiol, substituted or unsubstituted amino, Substituted or unsubstituted acyl, cyclic or acyclic, substituted or unsubstituted, branched or unbranched fatty acids Aliphatic, cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic, cyclic or acyclic, substituted or unsubstituted, branched or unbranched alkyl, cyclic or acyclic, Substituted or unsubstituted, branched or unbranched alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, aliphatic oxy, heteroaliphatic oxy alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, Aliphatic thioxy, heteroaliphatic thioxy, alkyl thioxy, heteroalkyl thioxy, Arylthioxy, heteroarylthioxy, mono- or di-aliphatic amino, mono- or di-hetero Heteroaliphatic amino, mono- or di-alkylamino, mono- or di-heteroalkylamino, mono- or di-arylamino, or mono- or di-heteroarylamino, or two R groups that combine to form a 5- or 6-membered heterocycle X1 Exemplary acyl groups include: The groups include aldehydes (-CHO), carboxylic acids (-CO2H), ketones, and halogenated groups. Includes acyls, esters, amides, imines, carbonates, carbamates and ureas Acyl substituents include, but are not limited to, those described herein that result in the formation of a stable moiety. The substituents described (e.g., aliphatic, alkyl, alkenyl, alkynyl, heteroaliphatic, poly heterocyclic, aryl, heteroaryl, acyl, oxo, imino, thioxo, cyano, iridium Socyano, amino, azido, nitro, hydroxyl, thiol, halo, aliphatic amino, Heteroaliphatic amino, alkylamino, heteroalkylamino, arylamino, heteroa Arylamino, alkylaryl, arylalkyl, aliphatic oxy, heteroaliphatic oxy alkyloxy, heteroalkyloxy, aryloxy, heteroaryloxy, Aliphatic thioxy, heteroaliphatic thioxy, alkyl thioxy, heteroalkyl thioxy, arylthioxy, heteroarylthioxy, acyloxy, etc., which are and may or may not be further substituted) .

[0104] The term "sulfonyl" refers to -SO2N(R bb)2, -SO2R aa , and -SO 2OR aa wherein R aa and R bb is defined herein That's right.

[0105] The term "aryl" refers to a group having 6 to 14 ring carbon atoms and provided in an aromatic ring system. A monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., 4n+2 aromatic ring system) without heteroatoms. For example, having 6, 10, or 14 π electrons shared within the cyclic array Radical ("C 6~14 In some embodiments, an aryl group is having 6 ring carbon atoms ("C6 aryl", e.g., phenyl). In this embodiment, an aryl group has 10 ring carbon atoms ("C 10 aryl”, e.g., 1 naphthyl, such as -naphthyl and 2-naphthyl. In some embodiments, the aryl group is , having 14 ring carbon atoms ("C 14 "aryl", e.g., anthracyl). "Aryl" also refers to an aryl ring as defined above which is substituted with one or more carbocyclyl or heterocyclyl groups. and includes ring systems in which the radical or point of attachment is on the aryl ring, in which case the carbon atom is The number of carbon atoms refers to the number of carbon atoms in the aryl ring system in succession. Each instance of an aryl group is independently unsubstituted (an "unsubstituted aryl") or contains one or more The substituents (e.g., -F, -OH, or -O(C 1~6 alkyl)) is substituted (" In certain embodiments, the aryl group is an unsubstituted C 6~14 Aryl In certain embodiments, the aryl group is a substituted C 6~14It is aryl.

[0106] The term "heterocyclyl" or "heterocyclic" refers to a ring having ring carbon atoms and 1 to 4 ring heterocycles. refers to a radical of a 3- to 14-membered non-aromatic ring system having heteroatoms, each heteroatom being independently and one or more selected from nitrogen, oxygen, and sulfur ("3- to 14-membered heterocyclyl"). For heterocyclyl groups containing a nitrogen atom, the point of attachment may be at a carbon atom or at a nitrogen atom, if valence permits. Heterocyclyl groups can be monocyclic ("monocyclic heterocyclyl") or polycyclic. Formula (e.g., fused, bridged, or spiro ring systems, e.g., bicyclic systems ("bicyclic heterocyclyl") ) or a tricyclic ring system (“tricyclic heterocyclyl”)), which may be saturated, or It may contain one or more carbon-carbon double or triple bonds. Heterocyclyl polycyclic ring systems include: One or both rings may contain one or more heteroatoms. The heterocyclyl ring as defined is fused to one or more carbocyclyl groups and the points of attachment are A ring system which is either a bicyclyl or heterocyclyl ring, or a heterocyclyl ring as defined above. The aryl ring is fused to one or more aryl or heteroaryl groups, and the point of attachment is at the heterocyclyl In such cases, the number of ring members in a sequence designates the number of ring members in the heterocyclyl ring system. Unless otherwise specified, each instance of heterocyclyl independently represents a non-substituted ring system. Heterocyclyl may be substituted ("unsubstituted heterocyclyl") or substituted with one or more substituents ("substituted heterocyclyl"). In certain embodiments, the heterocyclyl group is an unsubstituted 3- to 14-membered heterocyclyl. In certain embodiments, the heterocyclyl group is a substituted 3- to 14-membered heterocyclyl. In certain embodiments, heterocyclyl is a substituted or unsubstituted 3- to 7-membered heterocyclyl. cyclic heterocyclyl, where 1, 2 or 3 atoms of the heterocyclic ring system are substituted, as valences permit. , independently oxygen, nitrogen, or sulfur.

[0107] In some embodiments, a heterocyclyl group comprises ring carbon atoms and 1 to 4 ring heteroatoms. A 5- to 10-membered non-aromatic ring system having heteroatoms, each heteroatom being independently selected from nitrogen, oxygen, and and sulfur ("5- to 10-membered heterocyclyl"). A tetracyclyl group is a 5- to 8-membered non-aromatic ring having ring carbon atoms and 1 to 4 ring heteroatoms. A ring system in which each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-8 In some embodiments, a heterocyclyl group has ring carbon atoms and A 5- to 6-membered non-aromatic ring system having 1 to 4 ring heteroatoms, each heteroatom being independently In some embodiments, the heterocyclic group is selected from nitrogen, oxygen, and sulfur ("5- to 6-membered heterocyclyl"). In embodiments, the 5- to 6-membered heterocyclyl is selected from 1 to 3 nitrogen, oxygen, and sulfur. In some embodiments, the 5- to 6-membered heterocyclyl has a nitrogen, oxygen, or In some embodiments, the ring heteroatoms are selected from hydrogen and sulfur. A 5- to 6-membered heterocyclyl has one ring heteroatom selected from nitrogen, oxygen, and sulfur. do.

[0108] Exemplary 3-membered heterocyclyl groups containing 1 heteroatom include aziridinyl, Exemplary 4-membered heteroaryls containing one heteroatom include oxiranyl and thiiranyl. Tetracyclyl groups include azetidinyl, oxetanyl, and thietanyl. Exemplary 5-membered heterocyclyl groups containing 5 heteroatoms include tetrahydrofurane. dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl pyrrolyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclyl groups include dioxolanyl, oxathiolanyl, and Exemplary 5-membered heterocyclyls containing three heteroatoms include aryl, ... Groups include triazolinyl, oxadiazolinyl and thiadiazolinyl. Exemplary 6-membered heterocyclyl groups containing 1 heteroatom include piperidinyl, tetracycl ... Examples include thiahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclyl groups containing the group include piperazinyl, morpholinyl, dithiazinyl, and the like. Exemplary 6-membered heteroaryls containing three heteroatoms include anyl and dioxanyl. Cyclyl groups include triazinyl. Exemplary cyclyl groups containing one heteroatom include: Seven-membered heterocyclyl groups include azepanyl, oxepanyl and thiepanyl Exemplary 8-membered heterocyclyl groups containing one heteroatom include azocanyl, o- Exemplary bicyclic heterocyclyl groups include iodocanyl and thiocanyl. indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, thienyl tetrahydrobenzothienyl, tetrahydrobenzofuranyl, tetrahydroindolyl, tetrahydrobenzothienyl, tetrahydrobenzofuran ... Tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, decahydroquinolinyl Octahydroisoquinolinyl, octahydrochromenyl, octahydroisochromenyl, decahydro naphthyridinyl, decahydro-1,8-naphthyridinyl, octahydropyrrolo[3,2- b] Pyrrole, indolinyl, phthalimidyl, naphthalimidyl, chromanyl, chromenyl 1H-benzo[e][1,4]diazepinyl, 1,4,5,7-tetrahydropyranyl 5,6-dihydro-4H-furo[3,2-b]pyrrolyl, 6 ,7-Dihydro-5H-furo[3,2-b]pyranyl, 5,7-dihydro-4H-thieno [2,3-c]pyranyl, 2,3-dihydro-1H-pyrrolo[2,3-b]pyridinyl, 2,3-Dihydrofuro[2,3-b]pyridinyl, 4,5,6,7-tetrahydro-1H -pyrrolo[2,3-b]pyridinyl, 4,5,6,7-tetrahydrofuro[3,2-c] Pyridinyl, 4,5,6,7-tetrahydrothieno[3,2-b]pyridinyl, 1,2, 3,4-tetrahydro-1,6-naphthyridinyl and the like.

[0109] The term "heteroaryl" refers to an aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms. wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur. Monocyclic or polycyclic (e.g., bicyclic, tricyclic) 4n+2 aromatic ring systems (e.g., cyclic It refers to a radical of a quark (having 6, 10, or 14 pi electrons shared within the quark). 5-14 membered heteroaryl). In heteroaryl groups containing one or more nitrogen atoms, The point of attachment can be at a carbon atom or a nitrogen atom, valence permitting. The ring system may contain one or more heteroatoms in one or both rings. A heteroaryl ring as defined above fused to one or more carbocyclyl or heterocyclyl groups. and includes ring systems in which the point of attachment is on the heteroaryl ring, in which case the number of ring members is "Heteroaryl" also refers to the heteroaryl ring system as defined above. The heteroaryl ring is fused to one or more aryl groups and the point of attachment is on the aryl ring or on the heteroaryl ring. In such cases, the number of ring members may be fused polycyclic (aryl refers to the number of ring members in a (heteroaryl) ring system. aryl groups (e.g., indolyl, quinolinyl, carbazolyl, etc.); points of attachment are on one of the rings, for example, on a ring containing a heteroatom (e.g., 2-indolyl) or It may be present on either the ring containing the aryl group (e.g., 5-indolyl). In the formula, heteroaryl is a substituted or unsubstituted 5- or 6-membered monocyclic heteroaryl; One, two, three, or four atoms of the heteroaryl ring system are independently oxygen, nitrogen, or sulfur. In certain embodiments, heteroaryl is a substituted or unsubstituted 9- or 10-membered bicyclic heteroaryl. aryl, and 1, 2, 3, or 4 atoms of the heteroaryl ring system are independently selected from oxygen, It is nitrogen or sulfur.

[0110] In some embodiments, heteroaryl groups are provided at the ring carbon atoms and at the aromatic ring system. A 5- to 10-membered aromatic ring system having 1 to 4 ring heteroatoms, each heteroatom being independently In particular, nitrogen, oxygen, and sulfur ("5- to 10-membered heteroaryl"). In some embodiments, the heteroaryl group has 1 to 4 ring carbon atoms and 1 to 4 aromatic ring atoms provided on the aromatic ring system. A 5- to 8-membered aromatic ring system having ring heteroatoms, each heteroatom being independently selected from the group consisting of nitrogen, In some embodiments, the heteroaryl is selected from hydrogen, oxygen, and sulfur (a "5- to 8-membered heteroaryl"). In the present specification, a heteroaryl group is a group having 1 to 4 ring heteroatoms provided on the ring carbon atoms and an aromatic ring system. A 5-6 membered aromatic ring system having heteroatoms, each heteroatom being independently selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5- to 6-membered heteroaryl is selected from yellow ("5- to 6-membered heteroaryl"). Heteroaryl has 1 to 3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl is selected from nitrogen, oxygen, and sulfur. In some embodiments, the aryl group has 1 to 2 ring heteroatoms. The alkyl group has one ring heteroatom selected from nitrogen, oxygen, and sulfur. Unless otherwise specified, each instance of a heteroaryl group is independently unsubstituted ("unsubstituted heteroaryl"). Heteroaryl may be substituted with one or more substituents ("substituted heteroaryl"). In embodiments, the heteroaryl group is an unsubstituted 5-14 membered heteroaryl. In embodiments, the heteroaryl group is a substituted 5-14 membered heteroaryl.

[0111] Exemplary 5-membered heteroaryl groups containing one heteroatom include pyrrolyl, furacil, and cyclohexyl. Exemplary 5-membered heteroaryls containing two heteroatoms include phenyl and thiophenyl. The aryl group includes imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, Exemplary 5-membered heteroatoms containing three heteroatoms include thiazolyl and isothiazolyl. Aryl groups include triazolyl, oxadiazolyl and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include Exemplary 6-membered heteroaryl groups containing two heteroatoms include lysinyl. Examples include pyridazinyl, pyrimidinyl and pyrazinyl. Exemplary 6-membered heteroaryl groups containing 1,2,3,4-triazinyl and 1,2,3,4-triazinyl, respectively, include 1,2,3,4-triazinyl and 1,2,3,4-triazinyl. Exemplary 7-membered heteroaryl groups containing one heteroatom include azinyl. Examples include azepinyl, oxepinyl, and thiepinyl. Heteroaryl groups include indolyl, isoindolyl, indazolyl, benzotriazole, and benzotriazole. Zolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzisophenone Ranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzo benzothiazolyl, benzoisothiazolyl, benzothiadiazolyl, isothiazolyl Exemplary 6,6-bicyclic heteroaryl groups include benzophenone, benzophenone, benzothiazolinone, and benzothiazolinone. naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinolinyl, Exemplary tricyclic heteroaryls include benzoxalinyl, phthalazinyl, and quinazolinyl. Examples of the alkyl group include phenanthridinyl, dibenzofuranyl, carbazolyl, and acridinyl. , phenothiazinyl, phenoxazinyl and phenazinyl.

[0112] As used herein with respect to stereochemistry, the term "any" refers to the relative positions of the compounds. indicates that the specific or absolute stereochemistry has not been determined.

[0113] The terms "reduce," "reduced," "diminish," "inhibit," or "hinder" All are used herein to mean a statistically significant reduction. In embodiments, "reduce," "decrease," "reducing," "inhibiting," or "preventing" " typically refers to a decrease in a level of at least means a 10% decrease, e.g., at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least About 45%, at least about 50%, at least about 55%, at least about 60%, at least About 65%, at least about 70%, at least about 75%, at least about 80%, at least About 85%, at least about 90%, at least about 95%, at least about 98%, at least A reduction of about 99% or more may be included. "Inhibition" does not encompass complete inhibition or a reduction compared to a reference level. A reduction is 100% inhibition compared to the reference level. A reduction is preferably a reduction in the level of a given disorder. The decrease may be to a level that is considered within the normal range for a normal individual.

[0114] Treatment method Amyloids are present in the gut microbiota, including E. coli and several other proteobacteria. These microbial amyloids are produced in the digestive tract by members of the interact with the cells in contact with them, increasing α-synuclein expression and / or α-synuclein aggregation. The STC-1 cell line is derived from a tumor in the small intestine of a mouse and is a natural gastrointestinal It has many characteristics of secretory cells (McCarthy et al. (2015), STC -1 Cells.In:Verhoeckx K.et al.(eds)The I mpact of Food Bioactives on Health.Sprin ger, Cham.) Expression of α-synuclein in STC-1 cells was measured by Western blot analysis. In vitro assays measured by ELISA showed that the activity of wild-type CsgA-expressing E. coli strains was significantly increased. Exposure significantly increased α-synuclein expression, but not the isogenic csgA gene. Exposure to the gene variant had little effect on α-synuclein levels (e.g., See WO 2019 / 028456 and the references cited therein Thus, the exact mechanism by which CsgA affects α-synuclein expression remains unclear. However, CsgA interacts with enteroendocrine-like cells in the gastrointestinal tract and inhibits α-synuclein synthesis in vitro. It is possible that pathogenic microbial amyloids may cause overexpression of α-amyloids, which may disrupt the enteroendocrine secretion of the gastrointestinal tract. This suggests that similar effects may occur in vivo when contacted with cells or other cells. While mouse α-synuclein is not generally observed to aggregate, human α-synuclein Overexpression of ATP leads to aggregation, which in turn impairs cellular function and affects the gastrointestinal tract and enteric nervous system. It may propagate to neighboring cells in a prion-like manner and have deleterious effects on gastrointestinal function. These adverse effects include intestinal dysbiosis, increased intestinal permeability, and irritable bowel syndrome (IBS). IBS), inflammatory bowel disease (IBD), ulcerative colitis, or Crohn's disease These disorders include dysphagia, decreased intestinal motility, gastroparesis, constipation (chronic constipation and Constipation (including chronic idiopathic constipation), small intestinal bacterial overgrowth (SIBO), diarrhea, abdominal pain and / or Convulsions, bloating, flatulence, excessive salivation (hypersalivation), anorectal dysfunction, and impaired bowel coordination In some cases, the symptoms of amyloidosis may be associated with one or more of the following: Cardiovascular and gastrointestinal symptoms may be associated. For example, Parkinson's disease and parkinsonism ( In encephalomyelitis (a clinical syndrome characterized by tremor, bradykinesia, rigidity, and postural instability), dopamine Decreased levels can lead to the neurological symptoms of dyskinesia and the gastrointestinal symptoms of chronic idiopathic constipation Therefore, treatments that improve intestinal motility, including the methods of the present invention, can reduce dopamine levels in the digestive tract. This can improve stool absorption and thereby reduce dyskinesia. Treatments to manage intestinal dysbiosis (or, more generally, intestinal dysbiosis or intestinal hyperpermeability) include: , not only slows the progression of Parkinson's disease motor symptoms, but also The controlled "on-time" period can be increased.

[0115] Consistent with the ability of STC-1 cells to respond to E. coli CsgA in vitro, gastrointestinal cells It has been observed that they sense and respond to microbial amyloids. S. enterica CsgA inhibits mouse flu virus through activation of the TLR2 / PI3K pathway. Furthermore, U.S. Patent No. 9,814,756 has been shown to regulate gastrointestinal permeability of steroids. No. 11366666 A method for modulating gastrointestinal permeability through the administration of CsgA and / or CsgB mutants US Patent No. 9,814,756 discloses, inter alia, compounds having epithelial permeability reducing activity. Compositions comprising isolated curli fibrils, e.g., (i) 1 to 5 amino acids CsgA polypeptides that differ from naturally occurring CsgA polypeptides by substitutions, deletions, or additions. (ii) a gA polypeptide variant in which 1 to 5 amino acids are substituted, deleted, or added; CsgB polypeptide variants that differ from naturally occurring CsgB polypeptides in that they (iii) a combination of the CsgA polypeptide mutant and the CsgB polypeptide mutant; It is disclosed that administering the compound in combination reduces the permeability of the epithelium of the small intestine or large intestine. Therefore, there are additional mechanisms by which microbial amyloids interact with gastrointestinal cells. There may be more than one such mechanism, and at least one such mechanism is contemplated herein.

[0116] In one aspect, there is provided a method of inhibiting amyloid formation in a subject in need thereof, comprising: The present invention relates to a compound described herein, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. and administering to a subject in need thereof a therapeutically effective amount of a compound selected from the group consisting of acetaminophen, ...

[0117] In another aspect, the present invention provides a method for inhibiting, ameliorating, or potentially preventing amyloid disorders in a subject in need thereof. A method for reducing, delaying the onset of, treating or preventing idiopathic pulmonary edema comprising the steps of: (e.g., a compound of formula (I), or a compound of Table 1) or a pharmaceutically acceptable salt thereof. Such amyloid disorders include: , neurological disorders, as well as Parkinson's disease (PD), dementia with Lewy bodies, multiple system atrophy, and Developmental sclerosis (MS), frontotemporal dementia (FTD), REM sleep behavior disorder (RBD), α- Synucleinopathy, PD-related constipation, PD-related hypotension, Huntington's disease, Alexander disease Alzheimer's disease, amyotrophic lateral sclerosis (ALS), or Alzheimer's disease, and / or amyloid In some embodiments, the amyloid disorder includes intestinal disorders. Iosis, intestinal hyperpermeability, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), ulcerative Colitis or Crohn's disease.

[0118] In another aspect, a method of preventing or treating an inflammatory disorder in a subject in need thereof. Thus, the compounds described herein, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof In some embodiments, a method is provided herein, comprising administering to a subject , inflammatory disorders, bacterial sepsis, autoimmune diseases, lupus erythematosus, ischemia-reperfusion injury, In some embodiments, the disease is selected from stroke, metabolic disease, obesity-related metabolic inflammation, gout, and cancer. In the case of inflammatory disorders, the inflammatory disorder is lupus erythematosus.

[0119] In people with lupus erythematosus, e.g., systemic lupus erythematosus (SLE), the infection It is a common environmental trigger for flare (20-55%) and is associated with increased morbidity / mortality. Recombinant Salmonella typhimurium The curli-DNA complex activates innate / adaptive immunity and induces autoantibody formation. It acts as a potent immune stimulator by activating dendritic cells in vitro and In vivo activation in lupus-prone mice. Administration of recombinant curli-DNA complexes (50 μg, once or three times a week) to adult mice This leads to the formation of dsDNA and chromatin antibodies, which are signs of the onset of SLE. Infection with li-producing bacteria (E. coli or S. typhimurium) has also been shown to Promotes autoimmunity in lupus-prone mice. See, e.g., Gallo, PM, et al. See al. Immunity, 2015, 42, 1171-1184. These findings suggest that inhibitors of bacterial curli formation may prevent or inhibit autoimmune activation and may contribute to the development of This suggests that it may be a treatment for SLE in some cases.

[0120] In people with multiple sclerosis (MS), focal lymphocytic infiltration leads to myelin and axonal damage. It has been found to cause dysbiosis of the intestinal microbiota. For example, in people with MS, Akkermansia muciniphila muciniphila and Acinetobacter calcoaceticus Acter calcoaceticus levels increased, and Parabacter The level of ides distasonis decreases. For example, Cekanaviciu te,E.et al.Proc.Nat.Acad.Sci.,114,10713- See 10718. Additionally, in people with MS, Pseudomonas omonas, Mycoplana, Haemophilus ilus, Blautia and Dorea levels increase, Parabacteroides, Adlercroisia levels of ercreutzia and Prevotella spp. For example, Chen, J. et al. Nature Sci. Rep., 2016, 6 :28484. Also see Liu, JQ et al. J Exp Ne uropath.Exp.Neurol.2009,68,179 and Papado poulos D.et al.Mol.Cell Neurosci.2006,31 See, 597.

[0121] Synucleinopathy has been reported in the brains of deceased MS patients as well as in experimental autoimmune encephalomyelitis (E Patient-derived microbiota in EAE models has been observed in rat models. Transplantation of these mice was found to exacerbate disease severity (vs. transplantation of healthy controls). These findings suggest that inhibitors of bacterial curli formation may be a potential treatment for MS.

[0122] In some embodiments, the subject is screened for the presence of a marker in a gut sample (e.g., a fecal sample) of the subject. By detecting the presence and / or level of aggregates, the present composition can be used to A negative control (e.g., a healthy control subject or subject not having an amyloid disorder) may be selected. The presence or level of intestinal aggregates larger than those in a stool sample from a known control subject indicates that the subject In some embodiments, the subject may be in need of the composition. The presence and / or level of aggregates in a subject's intestinal sample, in combination with other factors such as sensitivity, The composition is selected as being in need thereof by detecting the presence of a marker.

[0123] In some embodiments, detecting the presence and / or level of intestinal aggregates in a sample from a subject The purpose is to identify bacterial proteins in the sample, e.g., curli-associated proteins such as CsgA. In some embodiments, detecting the presence and / or level of a protein in a subject Detecting the presence and / or level of intestinal aggregates in a sample may include determining the presence and / or level of amyloid in the sample. Detect levels of curli-associated proteins, such as CsgA, in bacteria that produce curli. For example, bacterial amyloid can be detected directly or by co-transfecting amyloid. nucleic acids that indicate the presence of amyloid-producing bacteria in the digestive tract of a subject. Examples of amyloid-producing bacteria include Csg-like bacteria such as Escherichia coli. Examples include the A-producing Enterobacteriaceae.

[0124] The compounds of formula (I) and their pharmaceutically acceptable salts can be administered in the form of a composition. In some embodiments, the composition can be found in any column of Table 3 or Table 4 (below). one or more compounds selected from the group consisting of compounds specified by the compound activity spectrum of "++" In some embodiments, the composition comprises or consists essentially of a compound. Compounds identified by a compound activity range of "+++" in any column of Table 3 or Table 4 The compound may comprise or consist essentially of one or more compounds selected from the group consisting of: In some embodiments, the composition is selected from the group consisting of "+++" and "++++" in any column of Table 3 or Table 4. one or more compounds selected from the group consisting of compounds specified by the compound activity spectrum In some embodiments, the composition comprises or consists essentially of a compound listed in Table 3 or Compounds identified by a compound activity range of "++" or "+++" in any column of Table 4 The compound may comprise or consist essentially of one or more compounds selected from the group consisting of: In some embodiments, the composition is identified by "++," "+++," or "A" in any column of Table 3 or Table 4. is selected from the group consisting of a set of compounds specified by a compound activity range of "++++" In some embodiments, the compound comprises or consists essentially of one or more compounds that The composition is identified by a "+", "++", "+++" or "+++" in any column of Table 3 or Table 4. +" one selected from the group consisting of a set of compounds specified by the compound activity spectrum In some embodiments, the composition comprises or consists essentially of the above compounds. The compound activity range of "+++" in the "αSyn ThT Assay" column of Table 3 or Table 4 is The compound may comprise or essentially comprise one or more compounds selected from the group consisting of compounds identified by In some embodiments, the composition comprises a "αSyn ThT" from Table 4. Compound activity ranges identified by a "++" or "+++" in the Assay column. comprising or consisting essentially of one or more compounds selected from the group consisting of In some embodiments, the composition comprises an "αSyn ThT Assay" from Table 3 or Table 4. A set of compounds identified by a compound activity range of "+", "++" or "+++" in the column The compound may comprise or consist essentially of one or more compounds selected from the group consisting of: In some embodiments, the composition is selected from the group consisting of the CsgA ThT assays listed in Table 3 or Table 4. 1 selected from the group consisting of compounds specified by the compound activity range of "++++" In some embodiments, the composition comprises or consists essentially of one or more compounds. The composition is a compound with an activity range of "+++" in the "CsgA ThT Assay" column of Table 3 or Table 4. or comprising one or more compounds selected from the group consisting of compounds identified by In some embodiments, the composition comprises a "Csg" from Table 3 or Table 4. Identified by a compound activity range of "+++" or "++++" in the "A ThT Assay" column The compound comprises or essentially comprises one or more compounds selected from the group consisting of the set of compounds In some embodiments, the composition comprises a "CsgA T" from Table 3 or Table 4. Compounds are characterized by a range of activity of "++," "+++," or "++++" in the "HT Assay" column. The compound comprises or essentially comprises one or more compounds selected from the group consisting of the set of compounds defined below. In some embodiments, the composition comprises a "CsgA" from Table 3 or Table 4. Compound activity ranges of "+", "++", "+++" or "++++" in the "ThT Assay" column The set of compounds identified by the group or consisting essentially of them.

[0125] The compounds of formula (I) and their pharmaceutically acceptable salts can be administered in the form of a composition. In some embodiments, the composition can be found in any column of Table 3 or Table 4 (below). One or more compounds selected from the group consisting of compounds specified by the compound activity ranges in In some embodiments, the composition comprises or consists essentially of a compound. Compounds identified by a compound activity range marked "***" in any column of Table 3 or Table 4 The compound may comprise or consist essentially of one or more compounds selected from the group consisting of: In this embodiment, the composition comprises a compound designated with "**" or "***" in any column of Table 3 or Table 4. The compound comprises one or more compounds selected from the group consisting of compounds defined by a bioactivity spectrum. In some embodiments, the composition comprises or consists essentially of a compound listed in Table 3 or Table 4. Compounds identified by a compound activity range of "**" or "***" in any column of The compound may comprise or consist essentially of one or more compounds selected from the group consisting of: In embodiments, the composition may be identified by a "**", "***" or "*" in any column of Table 3 or Table 4. Selected from the group consisting of the set of compounds specified by the compound activity range of "***" In some embodiments, the present invention comprises or consists essentially of one or more compounds. The composition is indicated by "*", "**", "***" or "****" in any column of Table 3 or Table 4. one or more compounds selected from the group consisting of a set of compounds defined by a compound activity spectrum In some embodiments, the composition comprises or consists essentially of a compound. Compound activity ranges identified by "****" in the "Reporter Assay" column of Table 3 or Table 4 The compound may comprise or consist essentially of one or more compounds selected from the group consisting of compounds In some embodiments, the composition comprises a compound selected from the group consisting of: One selected from the group consisting of compounds specified by the compound activity range of "***" In some embodiments, the composition comprises or consists essentially of the above compounds. The compounds are identified by "***" or "****" in the "Reporter Assay" column of Table 3 or Table 4. one or more compounds selected from the group consisting of a set of compounds specified by a range of In some embodiments, the composition comprises or consists essentially of a compound listed in Table 3 or Compound activity ranges marked with "**", "***" or "****" in the "Reporter Assay" column of Table 4 The set of compounds identified by the group In some embodiments, the composition comprises or consists essentially of a compound listed in Table 3 or Table 4. Compound activity marked with "*", "**", "***" or "****" in the "Reporter Assay" column one or more compounds selected from the group consisting of a set of compounds specified by a range of In some embodiments, the subject comprises or consists essentially of the intestinal By detecting the presence and / or level of aggregates in a sample (e.g., a fecal sample, etc.). Therefore, the present composition is selected as a necessary negative control (e.g., a healthy control subject, or stool samples from control subjects known not to have amyloid disorders) The presence or level of aggregates may indicate that the subject is in need of the composition.

[0126] In some embodiments, the composition has an IC of less than 150 μM. 50 of formula (I) In some embodiments, the present invention comprises or consists essentially of one or more compounds. Compositions with an IC of less than 80 μM 50 or comprising one or more compounds of formula (I) having the formula In some embodiments, the composition has an IC5 0. In some embodiments, the composition has an IC of less than 20 μM 50 one or more compounds of formula (I) In some embodiments, the composition comprises or consists essentially of a compound. IC<10 μM 50 comprising or essentially consisting of one or more compounds of formula (I) having the formula In some embodiments, the composition has an IC of less than 4.6 μM. 50 have In some embodiments, the compound comprises or consists essentially of one or more compounds of formula (I): In one embodiment, the composition has an IC 50 One or more compounds of formula (I) having the formula In some embodiments, the composition comprises or consists essentially of 1.3 IC of ~4.5 μM 50 comprising or essentially consisting of one or more compounds of formula (I) having the formula In some embodiments, the composition has an IC of less than 4.6 to 10 μM. 50 Some compounds may comprise or consist essentially of one or more compounds of formula (I) having the formula: In some embodiments, the composition has an IC of greater than 10 μM. 50 one or more compounds of formula (I) Comprising or consisting essentially of compounds.

[0127] In some embodiments, the composition exhibits a CsgA aggregation threshold of 125 in a ThT assay. % or less of Control B max % of one or more compounds of formula (I) In some embodiments, the composition comprises 100% or less of Control B. max %of Some compounds may comprise or consist essentially of one or more compounds of formula (I): In embodiments, the composition has a 90% or less Control B max % of one or more compounds of formula (I) In some embodiments, the composition comprises or consists essentially of a compound. Control B: 80% or less max % of one or more compounds of formula (I) In some embodiments, the composition comprises 70% or less of a control B max % Some compounds may comprise or consist essentially of one or more compounds of formula (I) having the formula: In some embodiments, the composition has a β-blocking activity of 60% or less of Control B. max % of formula (I) In some embodiments, the composition comprises or consists essentially of a compound of , 50% or less of control B max % of one or more compounds of formula (I) In some embodiments, the composition comprises 40% or less of a control B max % of one or more compounds of formula (I). In some embodiments, the composition has a control B max % of formula (I) In some embodiments, the composition comprises or consists essentially of the above compounds. is less than 20% of the control B max % of the compound of formula (I) In some embodiments, the composition contains 10% or less of % Control B. m ax % of one or more compounds of formula (I). In some embodiments, the composition has a control B max % of formula (I) In some embodiments, the present invention comprises or consists essentially of one or more compounds. The composition was 31 to 60% of Control B max % of one or more compounds of formula (I) In some embodiments, the composition comprises 11 to 31 % of Control B max % of one or more compounds of formula (I) In some embodiments, Control B max % is one or more compounds of formula (I) The results are determined against a vehicle control containing all components of the assay except for the In this embodiment, Control B max % was determined relative to a vehicle control containing solvent and reporter strain. In some embodiments, the solvent is DMSO. Assays and Controls (CsgA for ThT assay) is described in Example 26, and control B max % is paragraph [00033 6].

[0128] In some embodiments, the composition exhibits a 1 in a ThT fluorescence assay of CsgA aggregation. comprising or essentially comprising one or more compounds of formula (I) having a control AUC% of less than 25% In some embodiments, the composition has a control AUC% of 100% or less. Some compounds may comprise or consist essentially of one or more compounds of formula (I): In embodiments, the composition comprises one or more compounds of Formula (I) having a control AUC% of 90% or less. In some embodiments, the composition comprises or consists essentially of a compound of 8 0% or less of a control AUC%. In some embodiments, the composition has a control AUC% of 70% or less. In some embodiments, the compound comprises or consists essentially of one or more compounds of formula (I): In one embodiment, the composition comprises one or more compounds of formula (I) having a control AUC% of 60% or less. In some embodiments, the composition comprises 50% comprising or essentially comprising one or more compounds of formula (I) having the following control AUC%: In some embodiments, the composition comprises a compound of the formula having a control AUC% of 40% or less. (I). In this case, the composition contains one or more compounds of formula (I) having a control AUC% of 30% or less. In some embodiments, the composition comprises or consists essentially of 20% or less of and a compound of formula (I) having a control AUC% of In some embodiments, the composition comprises a compound of formula (I) having a control AUC% of 10% or less. In some embodiments, the compound comprises or consists essentially of one or more compounds of The composition comprises one or more compounds of formula (I) having a control AUC% of greater than 60%. In some embodiments, the composition comprises, or consists essentially of, 31 to 60 % of a control AUC% of one or more compounds of formula (I) In some embodiments, the composition has a control AUC% of 11 to 31%. Comprising or consisting essentially of one or more compounds of formula (I). In this embodiment, the control AUC% is calculated by comparing all components of the assay except for one or more compounds of formula (I). In some embodiments, the control AUC% is determined relative to a vehicle control containing and a vehicle control containing a reporter strain. is DMSO. The assay and controls are described in Example 26, and the AUC calculations are It is described in [000335].

[0129] In some embodiments, the composition comprises a compound of formula (I) that produces at least 10% inhibition In some embodiments, the compound comprises or consists essentially of one or more compounds of: The composition comprises one or more compounds of formula (I) that produce at least 20% inhibition. In some embodiments, the composition comprises at least 30 % inhibition of the IL-14 receptor agonist. In some embodiments, the composition comprises an inhibitor of formula (I) that produces at least 40% inhibition. In some embodiments, the compound comprises or consists essentially of one or more compounds of The composition comprises one or more compounds of formula (I) that cause at least 50% inhibition In some embodiments, the composition comprises at least 6 0% inhibition of the activity of the compound of formula (I) In some embodiments, the composition comprises a compound of formula (I) that causes at least 70% inhibition. In some embodiments, the compound comprises or consists essentially of one or more compounds of formula I). The compositions contain one or more compounds of formula (I) that cause at least 80% inhibition. In some embodiments, the composition comprises or consists essentially of at least 90% inhibition of the activity of one or more compounds of formula (I) In some embodiments, the composition comprises a compound of formula (I) that causes less than -10% inhibition. In some embodiments, the compound comprises or consists essentially of one or more compounds of The composition comprises one or more compounds of formula (I) that produce an inhibition of -10 to 10%. In some embodiments, the composition comprises 11 to 30 % inhibition of the IL-14 receptor agonist. In some embodiments, the composition comprises an inhibitor of formula (I) that produces 31 to 60% inhibition. In some embodiments, the compound comprises or consists essentially of one or more compounds of: The composition comprises one or more compounds of formula (I) that cause greater than 60% inhibition, or consists essentially of them.

[0130] In some embodiments, the subject is screened for the presence of a marker in a gut sample (e.g., a fecal sample) of the subject. By detecting the presence and / or level of aggregates, the present composition can be used to A negative control (e.g., a healthy control subject or subject not having an amyloid disorder) may be selected. The presence or level of intestinal aggregates larger than those in a stool sample from a known control subject indicates that the subject It may be indicated that the composition is in need.

[0131] Pharmaceutical Compositions, Formulations, Administration and Dosing In another aspect, a pharmaceutical composition comprising a compound described herein and a pharmaceutically acceptable carrier. The pharmaceutical compositions described herein inhibit amyloid formation. It is useful for

[0132] In certain embodiments, the pharmaceutical composition is formulated for delivery outside the systemic circulation of a subject. In certain embodiments, the pharmaceutical composition is formulated for delivery to the central nervous system of a subject. The composition may be formulated for enteral delivery and / or the composition may further be administered to the lower intestine or The composition may be formulated for controlled release in the colon or the rectum, and / or may be administered topically, orally, orally. The compositions may also be formulated for mucosal delivery. tablets, soft gels, spray-dried powders, polymer matrices, hydrogels, enteric-coated solids, crystalline solids, amorphous solids, glassy solids, coated particulates, liquids, sprays The mist may include a liquid, an aerosol, or a microcapsule.

[0133] In certain embodiments, the pharmaceutical composition is formulated for oral administration. In some cases, the pharmaceutical composition is formulated for administration by injection. Injections can be intravenous, subcutaneous, intramuscular, or intravenous. It may be intrameatal, intraperitoneal, intraspinal or intracranial.

[0134] Standard pharmaceutical and / or dietary supplement compounding techniques, e.g., Remington's The Science and Pra ctice of Pharmacy,21st Ed.Lippincott Wil The method described in Liams & Wilkins (2005) can be used. Accordingly, some embodiments provide for (a) a safe and therapeutically effective amount of any of the compounds described herein. or a pharmaceutically acceptable salt thereof, and (b) a pharmaceutically acceptable carrier. , a diluent, an excipient, or a combination thereof. include.

[0135] The pharmaceutical compositions comprise an effective amount of one or more compounds of the present invention. the desired biological and / or pharmacological effect of the compound, e.g., preventing or inhibiting the formation of amyloid aggregates. or inhibition, treatment or prevention of a neurological disorder or symptom of a neurological disorder, or treatment or prevention of a gastrointestinal disorder or symptom The compound (e.g., a compound described herein) is sufficient to achieve treatment or prevention. An "effective amount" or "effective dose" of a compound or a composition containing such a compound may be, for example, When delivered to a cell or organism according to the selected dosage form, route and / or schedule "Effective amount" refers to an amount sufficient to achieve a desired biological and / or pharmacological effect. and "therapeutically effective amount" may be used interchangeably. The absolute amount of a particular compound or composition that is effective may vary depending on the desired biological or pharmacological endpoint. These may vary depending on factors such as the agent being delivered, the target tissue, etc. Those skilled in the art will appreciate that various embodiments may be used. In certain embodiments, an "effective amount" may be administered to a subject in a single dose or by the use of multiple doses. will understand more.

[0136] "Administering" includes its conventional and usual meaning as understood by those skilled in the art in light of this disclosure. It refers to providing a subject with a pharmaceutical product, dietary supplement, or composition, This includes, but is not limited to, administration by a medical professional and self-administration. The compounds or pharmaceutically acceptable salts thereof may be administered by any of a variety of methods, including, but not limited to, oral, intraperitoneal, or intravenous routes. Any of the accepted modes of administration for drugs with similar efficacy, such as intracavitary or rectal Oral administration may be by way of administration of the composition that is the subject of a preferred embodiment. However, in some embodiments, the administration according to the methods of the present disclosure The composition is administered rectally, such as by enema or suppository. Administration of the compounds can be performed extracorporeally, for example, by apheresis or dialysis.

[0137] The term "drug" shall be used in its conventional and usual manner as understood by those skilled in the art in light of this disclosure. It means any substance, molecule, element, compound, entity, or combination thereof. This includes, for example, proteins, polypeptides, peptides or mimetics, organic small molecules, polysaccharides, polynucleotides, polymers, resins, organic or inorganic particulates, organic or inorganic These include, but are not limited to, natural products, synthetic compounds, young It may be a compound or a chemical compound, or a combination of two or more substances.

[0138] The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" is used in the context of this disclosure. has its customary and ordinary meaning as understood by those skilled in the art in light of the and any solvents, diluents, emulsifiers, binders, buffers, dispersion media, coatings, antimicrobial agents and and antifungal agents, isotonic and absorption delaying agents, etc., which are known to be useful in the preparation of pharmaceutical formulations. The present invention also includes any other such compounds known to those skilled in the art. The use of such media and agents is well known in the art. Except insofar as it is incompatible with the active ingredient, its use in therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions. Various commonly used adjuvants may also be included. Such compounds can be found in the literature, e.g., the Merck Index, Merck & Co. Any, Rahway, NJ. For further considerations, see, for example, Gilman et al. (Eds.) (1990), odman and Gilman's:The Pharmacological Basis of Therapeutics,8th Ed.,Pergamon P It is described in ress.

[0139] In accordance with the methods and compositions of some embodiments herein, a pharmaceutically acceptable carrier or Some examples of substances that can serve as components are lactose, glucose and sucrose. sugars such as sucrose; starches such as corn starch and potato starch; Cellulose such as sodium carboxymethylcellulose, ethyl cellulose, and methyl cellulose Tragacanth powder; malt; gelatin; talc; stearic acid and stearic acid Solid lubricants such as magnesium phosphate; calcium sulfate; peanut oil, cottonseed oil, sesame oil, Vegetable oils such as olive oil, corn oil and theobroma oil; propylene glycol, Polyols such as glycerin, sorbitol, mannitol and polyethylene glycol alginic acid; emulsifiers such as TWEENS; wetting agents such as sodium lauryl sulfate; coloring Flavoring agent; Tableting agent; Stabilizer; Antioxidant; Preservative; Pyrogen-free water; Isotonic sodium chloride water; and / or phosphate buffer, or any combination thereof.

[0140] Pharmaceutically acceptable compounds for use in conjunction with one or more of the compounds for administration described herein The choice of possible carrier may be determined by the way the compound is to be administered.

[0141] Additionally, the present disclosure provides various salts, esters, hydrates, prodrugs, and derivatives of the compounds described herein. This includes compositions containing isotopically substituted analogs such as hydroxybenzoates, fluorinated analogs, or deuterated forms.

[0142] The term "pharmaceutically acceptable salt" means, within the scope of sound medical judgment, a salt that is free from excessive toxicity, Suitable for use in contact with human and lower animal tissues without irritation, allergic reactions, etc. A pharmaceutically acceptable salt is a salt that is suitable for use in a pharmaceutical composition that has a reasonable benefit / risk ratio. It is well known in the art, for example, Berge et al., incorporated herein by reference. J.Pharmaceutical Sciences,1977,66,1-19 Pharmaceutically acceptable salts of the compounds of the present invention are described in detail in the following. Suitable salts include those derived from suitable inorganic and organic acids and bases. Examples of acceptable non-toxic acid addition salts include those from hydrochloric, hydrobromic, phosphoric, sulfuric and perchloric acids. Inorganic acids, or acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malo Salts of amino groups formed with organic acids such as carboxylic acids or ion exchange methods known in the art. Other pharmaceutically acceptable salts of amino groups formed by other known methods. Acceptable salts include adipate, alginate, ascorbate, and aspartate. Salt, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphor, camphor -sulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl Sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerol Phosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2 -hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalene Sulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, Pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrin acid salts, pivalates, propionates, stearates, succinates, sulfates, tartrates, Examples include thiocyanate, p-toluenesulfonate, undecanoate, and valerate. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium, Representative alkali metal salts or alkali metal salts include ammonium and N+(C1-4 alkyl)4- salts. Alkali earth metal salts include sodium, lithium, potassium, calcium, magnesium, Further pharmaceutically acceptable salts include, where appropriate, halides, Hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates and alkyl Non-toxic ammonium, quaternary ammonium salts formed using counterions such as arylsulfonates Examples include ammonium and amine cations.

[0143] "Solvate" has its customary and ordinary meaning as understood by those of skill in the art in light of this disclosure. It has the following properties: the interaction of the solvent with the active pharmaceutical ingredient (or API), metabolites, or salts Suitable solvates include pharmaceutically acceptable solvates, including hydrates. It is a solvate.

[0144] As used herein, "systemic circulation" is understood by those skilled in the art in light of this disclosure. It has its customary and ordinary meaning. It refers to circulation within the blood or circulatory system of a subject.

[0145] As used herein, "enteric coating" refers to a coating that is readily absorbed by the body of a subject, as would be understood by one of ordinary skill in the art in light of this disclosure. It has its customary and ordinary meaning as understood. by controlling the absorption of lactic acid, for example by providing resistance to stomach acid, or by inhibiting the growth of having a higher solubility at neutral or basic pH, allowing it to reach the desired portion of the gastrointestinal tract Coating or dissolving particles increases the likelihood that the particles will be protected from the solvent until the refers to pharmaceutical excipients that are placed around the particles. Typical enteric coatings include For example, Remington's The Science and Practice of Pharmacy,21st Ed.Lippincott Williams&Wi Exemplary enteric coatings include those described in L.K.K.N. (2005). Contains shellac, sodium alginate, zein, cellulose acetate trimellitate , methyl methacrylate-methacrylic acid copolymer, polyvinyl acetate phthalate, polylactic acid, Polylactic-co-glycolic acid, hypromellose acetate, hypromellose acetate succinate, Hydroxypropylmethylcellulose phthalate, cellulose acetate succinate, cellulose acetate phthalate Cellulose, methyl acrylate-methacrylic acid copolymer, polyvinyl acetate phthalate, Op adry®, and others known in the art of drug delivery and formulation. According to some embodiment methods and compositions, the compounds described herein may include, but are not limited to, The compositions comprising the described compounds further comprise an enteric coating.

[0146] The term "gut-selective" as used herein is understood by those of skill in the art in light of this disclosure. It has its customary and ordinary meaning as being released into the digestive tract of a subject and is preferably Refers to compositions or preparations that are not absorbed or, if absorption occurs, do not enter the systemic circulation. vinegar.

[0147] As used herein, the term "essentially enteric" is understood by those of skill in the art in light of the present disclosure. With respect to pharmaceutical preparations, the term "disintegration" has its customary and ordinary meaning as understood in the gastric environment. It refers to a composition that has the ability to prevent breakdown or release.

[0148] The compositions described herein for administration to a subject are preferably provided in unit dosage form. As used herein, "unit dosage form" is understood by those of skill in the art in light of this disclosure. has its customary and ordinary meaning. It is used in accordance with good medical practice to refers to a composition containing a quantity of a compound suitable for administration to an animal, provided that the preparation of a single or unit dosage form is This does not imply that the dosage form is to be administered once per day or once per course of treatment. A unit dosage form is one in which several unit dosage forms are administered throughout the day to complete the daily dose. According to the present disclosure, a unit dosage form may contain a single daily dose or divided doses to be administered. It may be given more or less frequently than once a day and may be given multiple times during the course of treatment. Such dosage forms may be administered orally, rectally, nasally and / or parenterally, as appropriate for their formulation. Administration can be by any mode, including orally. Although single doses are specifically contemplated, The compositions administered according to the methods described in may also be administered in the form of a continuous infusion or as an implantable injection. It can also be administered via an infusion pump.

[0149] The methods described herein may be used in conjunction with various routes of administration, for example, oral, nasal, rectal, or Any of a variety of suitable forms for the parenteral route of administration may be utilized. Depending on the route, a variety of pharmaceutically acceptable carriers well known in the art can be used. Pharmaceutically acceptable carriers include, for example, solid or liquid fillers, diluents, hydrogels, These include tropes, surfactants and encapsulating substances. Optional pharmaceutically active materials may be included as long as they do not interfere in quality. The amount of carrier used should be such that it provides a practical amount of material for administration per unit dose of the compound. The techniques and methods for making dosage forms useful in the methods described herein are The compositions are described in the following references: Modern Pharmaceutics, 4 th Ed.,Chapters 9 and 10(Banker&Rhodes,e ditors, 2002), Lieberman et al., Pharmaceut ical Dosage Forms: Tablets (1989) and Ansel ,Introduction to Pharmaceutical Dosage F orms 8th Edition (2004), all of which are referenced and is incorporated herein by reference.

[0150] Various oral dosage forms, including solid forms such as tablets, capsules, granules, and / or bulk powders The tablets may contain suitable binders, lubricants, diluents, disintegrants, coloring agents, flavoring agents, etc. Compressed, powdered, enteric coated tablets containing flavorings, flow inducers, and / or melting agents coated, sugar-coated, film-coated, or Further solid dosage forms include milled powders, spray dried powders, crystalline powders, and the like. and may be administered as a tablet. are administered as aerosols or airborne particles, e.g., for nasal or pulmonary delivery. The liquid oral dosage form may contain suitable solvents, preservatives, emulsifiers, suspending agents, diluents, sweeteners, Non-effervescent granules containing melting agents, coloring agents and / or flavoring agents, or any combination thereof Aqueous solutions, emulsions, suspensions, solutions and / or suspensions, and effervescent Further liquid dosage forms include effervescent preparations reconstituted from granules. Such dosage forms may include intranasal injection, nasal wash, lung wash, spray or aerosol. It may contain a liquid for delivery.

[0151] Preparation of unit dosage forms for oral administration according to the methods and compositions of some embodiments herein Pharmaceutically acceptable carriers suitable for formulation are well known in the art. Tablets are typically Conventional pharmaceutically compatible adjuvants include calcium carbonate, sodium carbonate, mannitol, inert diluents such as starch, gelatin and / or cellulose; binders such as sucrose; starch, alginic acid and / or croscarmellose Disintegrants: magnesium stearate, stearic acid, microcrystalline cellulose, carboxymethyl Tablets may also contain lubricants such as cellulose and / or talc. stabilizing agents, e.g., poloxamer, cremophor / Kolliphor® / Lutrol®, or methylcellulose, hydroxypropyl methylcellulose The diacid may include benzoyl benzoates, ... Glidants such as silicon dioxide can be used to improve the flow characteristics of the powder mixture. For example, FD&C dyes may be added for appearance. phosphorus, menthol, peppermint, and / or fruit flavors, or any of these Sweeteners and flavoring agents, such as any combination thereof, are useful adjuvants for chewable tablets. The formulation typically includes one or more solid diluents as disclosed above. The selection of the carrier component depends on the taste, It depends on secondary considerations such as cost and storage stability, which can be easily determined by one skilled in the art. can be done.

[0152] Oral (PO) compositions according to some embodiments of the methods and compositions herein also include liquid pharmaceutically acceptable salts suitable for the preparation of such compositions, including aqueous solutions, emulsions or suspensions. Suitable carriers are well known in the art. Syrups, elixirs, emulsions and / or Typical components of carriers for suspensions are ethanol, glycerol, propylene glycol, etc. Examples of suitable glycerols include ethanol, polyethylene glycol, liquid sucrose, sorbitol, and / or water. For suspensions, typical suspending agents are methylcellulose, carboxymethylcellulose, Sodium cellulose, AVICEL RC-591, tragacanth and / or alginate Typical wetting agents include sodium phosphate, and lecithin and / or polysorbate 80, and typical preservatives include methylparaben and / or sodium benzoate. The oral liquid composition may also include any of the above-disclosed The composition may contain one or more ingredients such as sweeteners, flavoring agents and / or coloring agents, as described in .

[0153] Such compositions also can be applied by conventional methods, typically with pH or time dependent coatings. The subject compounds may be coated with a coating, so that one or more of the subject compounds are coated as desired. The drug is released into the gastrointestinal tract either close to application or at various times to prolong the desired effect. Exemplary dosage forms for release in the gastrointestinal tract include cellulose acetate phthalate, polyacetate phthalate, and Polyvinyl, Hydroxypropylmethylcellulose Phthalate, Ethylcellulose, Eudra coatings, waxes, alginates and / or shellac, or other coatings known to those skilled in the art. or any combination thereof. In some embodiments, the compositions administered in accordance with the methods described herein can In some embodiments, the compound is formulated for release in the intestinal tract. The compositions administered are formulated for release in the lower gastrointestinal tract. The composition may be in the form of an enteric coated capsule, tablet, softgel, or essentially an enteric coated capsule. It is provided as a rule.

[0154] The actual unit dose of the composition according to the methods and compositions of some embodiments herein is In some embodiments, the amount of 1 kg of body weight of the subject in the formulation is dependent on the one or more compounds in the formulation. Doses in milligrams per gram range from 0.01 mg / kg to 0.05 mg / kg per day. g / kg, 0.04mg / kg per day to 0.1mg / kg per day, 0.09m g / kg to 0.15 mg / kg, 0.14 mg / kg to 0.2 mg / kg per day , 0.2mg / kg to 0.5mg / kg per day, 0.4mg / kg to 1mg / kg, 1mg / kg per day to 6mg / kg, 5mg / kg per day to 500 mg / kg or more per day, 10 mg / kg or less per day to 70 mg / kg or less per day 50 mg / kg to 80 mg / kg, 70 mg / kg to 120 mg / kg per day; 100mg / kg to 300mg / kg per day, or 250mg / kg per day In some embodiments, the dose may be 10 to 500 mg / kg per day. Less than 0mg / kg, Less than 500mg / kg, Less than 300mg / kg, Less than 200mg / kg More than, less than 150mg / kg, less than 100mg / kg, less than 50mg / kg, 40mg / kg Less than g, Less than 30mg / kg, Less than 25mg / kg, Less than 20mg / kg, 10mg / kg Less than 7.5mg / kg, less than 6mg / kg, less than 5mg / kg, less than 4mg / kg less than 3 mg / kg, less than 2.5 mg / kg, or less than 1 mg / kg, or The amount may be within a range defined by any two of the amounts. Actual unit doses are 5, 10, 25, 50, 75, 100, 150, or 200 mg / kg, or an amount within a range defined by any two of the foregoing amounts. Thus, for example, the dosage range for a 70 kg person would be 0.1 mg to 1 m g, 0.9mg to 2mg, 1.5mg to 5mg, 4mg to 10mg, 9mg to 2 0mg, 15mg to 50mg, 40mg to 75mg, 50mg to 100mg, 75 mg to 200mg, 100mg to 300mg, 200mg to 400mg, 350m g to 750mg, 500mg to 1g, 750mg to 2g, 1g to 5g, 2.5g to 6g, 4g to 10g, 8g to 20g, 15g to 35g, or 1g or less to 35g g or more, or within a range defined by any two of the preceding amounts. In embodiments, the actual unit dose is 6 g. In some embodiments, the actual unit dose is In some embodiments, the actual unit dose is 35 g. In embodiments, the actual unit dose is 1 g or less, but not 0. In some embodiments, the actual unit dose is 10 g or less, but not zero. The actual unit dose is 35 mg or less, but not zero.

[0155] As used herein, a "loading dose" refers to an initial dose of a compound that is higher than subsequent doses. .

[0156] As used herein, a "maintenance dose" refers to a subsequent dose that follows a loading dose and is Those skilled in the art will appreciate that the dosage form or mode of administration of the maintenance dose will be used for the loading dose. It is understood that the actual results may differ from those shown in any of the embodiments disclosed herein. In this context, the maintenance dose may be administered in accordance with any dosing schedule contemplated herein (e.g., monthly or Multiple times, once or more every two weeks, once or more times a week, once or more times a day may include administering the unit dosage form multiple times (including, but not limited to, multiple times). It is contemplated that a drug holiday period may be incorporated into the administration of the maintenance dose. A suitable withdrawal period may occur immediately after administration of the loading dose or at any time during maintenance dosing. As used herein, the period during which maintenance doses are administered is referred to as the "maintenance phase" of the treatment period. It can be called.

[0157] As used herein, "mode of administration" refers to the means by which one or more compounds are administered to a subject. As used herein, "mode of administration" refers to the dosage form (e.g., tablet, powder, solution, suspension, emulsion, etc.) and the mechanism by which the dosage form is administered to a subject (e.g., by injection, topically, e.g., by cream, lotion, or patch; orally via oral suspension, oral film, or mouth rinse). When used, "mode of administration" also refers to the dose, dosage, and administration rate at which a compound is administered to a subject. Includes a feeding schedule.

[0158] In some embodiments, the compositions administered according to the methods of the present disclosure are prepared as food, beverage, or The may be provided with or mixed into other ingestible products. In some embodiments, the beverage, food, or other ingestible item may be candy, applesauce, yogurt, or the like. Yogurt, soft pudding, gelatin foods, juice, milk, soy or nut drinks, may include one or more of a thickened beverage, or cheese, or any combination thereof Those skilled in the art will appreciate that the combination of compositions administered according to the methods of the present disclosure may be administered by ingestion of the compositions. It is easy to understand that it can be combined with any suitable food or beverage to make it easier to eat. You will understand.

[0159] In some embodiments, administration according to the methods and compositions of some embodiments herein The regimen includes administering a loading dose followed by a maintenance dose. The loading dose is 20g or less but not 0, 15g or less but not 0, 10 g or less but not 0, 6g or less but not 0, 4g or less but not 0 , 2g or less but not 0g, or 1g or less but not 0g, or any of the aforementioned amounts In some embodiments, the maintenance dose is an amount within a range defined by any two of: 20g or less but not 0, 10g or less but not 0, 6g or less but not 0 4g or less but not 0, 2g or less but not 0, 1g or less but Not 0, 500 mg or less but not 0, or 250 mg or less but not 0 or an amount within a range defined by any two of the foregoing amounts.

[0160] In some embodiments, according to the methods and compositions of some embodiments herein, The loading dose is administered over a daily or 24 hour period. The loading dose is administered in a single dose. In some embodiments, the loading dose is administered in multiple doses. In some embodiments, the loading dose is administered multiple times in a single day or 24 hour period. In some embodiments, the loading dose is administered over two days. In some embodiments, the loading dose is administered over a 3 day period. In some embodiments, the loading dose is administered over a 4 day period. In some embodiments, the loading dose is administered over a period of 5, 6, or 7 days. is administered for a period of 8 to 14 days or less. The loading dose is administered over 14 days.

[0161] The methods according to the present disclosure may be used to enhance the efficacy of any administered treatment, including those described herein. It is contemplated that the timing of administration of the composition may be varied or controlled. In some cases, compositions administered according to the methods of the present disclosure may be administered with food, e.g., at mealtime or mealtime. In some further embodiments, the present disclosure provides a method for administering a medicament for the treatment of a malaria parasite, such as fluticasone, fluticasone, or fluticasone-containing fluticasone, simultaneously with other ingestion of the medicament. The compositions administered according to the method of claim 1 may be administered immediately before or after a meal or other ingestion of food. In some further embodiments, the compositions administered according to the methods of the present disclosure The items are taken within 1-5 minutes, 3-10 minutes, 6-15 minutes before or after a meal or other intake of food. Administer within 10-20 minutes, within 15-30 minutes, within 20-45 minutes, or within 1 hour In some embodiments, the compositions administered according to the methods of the present disclosure , for example, 1 to 3 hours before or after, 2 to 5 hours before or after, 4 to 6 hours after, or a meal or other ingestion of food. 8 hours before or after, 6 to 12 hours before or after, 9 to 18 hours before or after, 12 to 24 hours before or after It can be administered without food, or more than 24 hours before or after.

[0162] As used herein, the "period of treatment" begins with the administration of the first dose and ends with the administration of the last dose. This refers to the time it takes for the dose to be administered and for the duration of the treatment to be completed, which is related to intestinal hyperpermeability or "leaky gut." The condition of the subject being treated therefor by a person skilled in the art of treating the neurological disorder or disorder to which the Such duration is determined with reference to the patient's physical condition and health status. or periodic, sporadic changes in amyloid levels known to those skilled in the art of treating neurological disorders. The determination may be made with reference to periodic or continuous monitoring.

[0163] As used herein, a "drug holiday" refers to a period of 24 hours or more during which Either no dose is administered to the elephants or a reduced dose is administered to the subjects. As used herein, a "reduced dose" refers to the total daily dose that is to be administered to a subject. Refers to a dose less than

[0164] According to the present disclosure, the administration schedule can be modified to achieve the desired therapeutic effect. In each embodiment disclosed herein, the change in the administration schedule can be implemented. The present invention may be repeated throughout the course of a treatment protocol. In this case, the first dose may be greater than, less than, or the same as the dose after the first dose. In each of the embodiments disclosed herein, the loading dose may be any of the disclosed dosages. The regimen may be preceded by a drug holiday period which may or may not follow the administration of a loading dose.

[0165] In some embodiments, the methods of the present disclosure comprise administering to a subject a subject one or more of the compositions provided herein. daily or less frequently than daily, for example, every 2 days, every 3 days, every 4 days, every 5 days, every 6 days every seven days, or every seven days, or within a range defined by any two of the aforementioned times. In some embodiments, the compositions described herein include administering the compositions over a period of time. is formulated for such administration.

[0166] According to the methods disclosed herein, treating or inhibiting a disorder involving amyloid formation includes administering to a subject a therapeutically effective amount of a compound selected from the group consisting of steroids, steroid hormone ... and steroid hormones. This can be achieved by adjusting the dosing schedule for administration of the composition, so that After experiencing an intermittent, partial, or complete reduction in medication over a period of time, the subject is given In some embodiments, the dose is administered daily for 1 to 30 days. In some embodiments, the drug-free period is 1 to 30 days. In some further embodiments, the compositions of the present disclosure are administered in the following manner: In some other embodiments, the drug may be completely cleared from the subject's body before administration of the drug. In some further embodiments, a dose less than the usual daily dose is administered throughout the period. It is possible that sub-therapeutically effective amounts of the administered composition remain in the subject during a drug holiday period. In some further embodiments, the administered composition is maintained at a therapeutic level in the affected tissue. In some embodiments, the composition is capable of remaining in the subject in a sufficient amount to sustain the intended effect. The subject is a patient with amyloidosis at any time after the onset of one or more of the aforementioned symptoms of a neurological disorder associated with amyloid formation. In some embodiments, the compositions according to the methods described herein are administered in one or more In some embodiments, the compound described herein is administered prior to the onset of symptoms of the above disorders. The compositions according to the methods described above are administered simultaneously with or after the onset of symptoms of one or more of the above disorders. It is given.

[0167] How to use The present disclosure provides methods for inhibiting, ameliorating, reducing the likelihood, delaying the onset, and treating amyloid disorders. The present invention provides a method for the treatment and / or prevention of bacterial infections, comprising: (1) treating bacterial infections on the surface of bacteria or in the proximal extracellular space; (2) bacterial amyloid aggregation in the gastrointestinal tract or olfactory system (including enteroendocrine cells and enteric neurons) ) the interaction of bacterial amyloid with α-synuclein in the gut, and / or (3) the gastrointestinal tract Aggregation of α-synuclein in enteroendocrine cells and enteric neurons This includes methods of inhibiting or disrupting the above.

[0168] According to the methods of the present disclosure, α-synuclein is derived from amyloid precursor protein (APP). Amyloid, medin, tau, apolipoprotein AI, atrial natriuretic factor, beta Amyloid, cystatin, IAPP (amylin), beta-2 microglobulin, Transthyretin, PrP, gelsolin, lysozyme, huntingtin, keratoepithelin (Keratoepithelin), calcitonin, prolactin, serum amyloid A , superoxide dismutase 1 (SOD1) and / or immunoglobulin light chain AL cyst Amyloids representative of a wider range of known host amyloid proteins, including one or more of The compositions and methods disclosed herein should be considered as a single If an amyloid protein (bacterial or human) promotes the aggregation of another amyloid protein, The compounds can be adapted by one skilled in the art to interfere with the aggregation of any amyloid protein.

[0169] Without being limited by theory, it is believed that the effects of the oxidative stress on the oxidative stress may be inhibited or prevented using the methods of the present disclosure. Representative disorders that exhibit amyloid formation and proteins involved in these disorders include: , including, but not limited to, those disclosed in Table 2. [Table 2]

[0170] The compositions and methods of the present invention also provide a method for treating intestinal dysbiosis, intestinal hyperpermeability, irritable bowel, and the like. IBS, inflammatory bowel disease (IBD), ulcerative colitis and / or Crohn's disease , can be used to treat amyloid-mediated disorders of the gastrointestinal tract. Symptoms include dysphagia, decreased intestinal motility, gastroparesis, constipation (including chronic constipation and chronic idiopathic constipation) ), small intestinal bacterial overgrowth (SIBO), diarrhea, abdominal pain and / or cramps, bloating, flatulence, saliva One that includes hypersalivation (excessive salivation), anorectal dysfunction, bowel coordination disorder, and nausea May be associated with the above symptoms.

[0171] As used herein, the term "intestinal dysbiosis" means, in light of this disclosure, has its conventional and ordinary meaning as understood by those skilled in the art, and includes the digestive tract or intestines, particularly the small intestine Dysbiosis refers to an imbalance and / or maladaptation of the bacterial flora or microflora within the body. changes in the composition of the intestinal or gastrointestinal microbiota in terms of the species / strains present and / or the presence It is characterized by a change in the relative abundance or proportion of species / strains that are involved in the host organism. The adverse effects on the host organism are due to microbiota-mediated electrolyte balance changes in the lance, biofilm formation, the integrity of the barrier formed by the intestinal epithelial lining, or , directly (e.g., as a toxicant or effector) or indirectly to the health of the host. the production of harmful metabolites (e.g., as precursors to toxins or effectors) by the microbiota Emission can result from:

[0172] As used herein, the term "intestinal permeability enhancement" refers to the ability of a subject to be stimulated by an intestinal permeability enhancing agent to enhance intestinal permeability. This has its customary and ordinary meaning as understood by the practitioner. This refers to an abnormally increased permeability of the barrier formed by the intestinal epithelial lining between the Permeability may be due to inflammation of the intestinal lining and / or failure of tight junctions between cells of the intestinal epithelium, which This allows material to pass from the lumen into the surrounding tissue, some of which may enter the peritoneal cavity and / or This leakage of substances from the digestive tract or intestinal lumen can enter the systemic circulation. This is sometimes called "leaky gut" or "leaky gut syndrome."

[0173] As used herein, the term "amyloid disorder" (a variation of this root term) forms) are associated with any or all of the disorders in Table 2, as well as amyloid-mediated disorders of the gastrointestinal tract Including, but not limited to:

[0174] As used herein, "mammalian amyloid or mammalian amyloid precursor" refers to a compound that is amyloid-containing compound. The term "tau," "beta-amyloid derived from amyloid precursor protein," "medin," Apolipoprotein AI, atrial natriuretic factor, beta-amyloid, cystatin, IAPP (amylin), beta-2 microglobulin, transthyretin, PrP, genomic DNA Lusoline, lysozyme, huntingtin, keratoepithelin in), calcitonin, prolactin, serum amyloid A, and / or immunoglobulin L The specific amino acid sequences disclosed herein include, but are not limited to, one or more of the amino acid sequences A, B, C, D, E, E, F ... In the methods and compositions, the microbial amyloid or microbial amyloid precursor is CsgA. Includes:

[0175] The terms "amyloid aggregate" and "aggregate of amyloid proteins" are used interchangeably. It is used.

[0176] Some embodiments inhibit, ameliorate, reduce the likelihood of, or prevent amyloid disorders. and methods for delaying the onset, treating, or preventing the onset of a disease, the methods comprising administering a composition described herein. The amyloid disorder includes administering to a subject in need thereof an α-synucleinopathic drug. Qi, Parkinson's disease, Lewy body dementia, incident Lewy body disease, Alzheimer's Lewy body variant of idiopathic encephalopathy, multiple system atrophy, and / or pure autonomic failure The amyloid disorder may be selected from the group consisting of any combination thereof. , intestinal hyperpermeability, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), ulcerative colitis or In some embodiments, the compound administered in this method may be selected from the group consisting of Crohn's disease. The composition may comprise, consist essentially of, or consist of any of the compounds in Table 1. In some embodiments, the composition administered in this method comprises a compound described herein. Some embodiments of the method include, consist essentially of, or consist of a compound. In some cases, the amyloid disorder involves intestinal amyloid aggregates. For example, the aggregates are caused by bacterial tumors. Proteins may include curli-associated proteins such as CsgA. In some embodiments, the method comprises determining the presence of such bacterial proteins in an intestinal sample of a subject. the presence or level of nucleic acids encoding microbial (e.g., bacterial) proteins; or levels of bacterial proteins (e.g., C) in a subject's intestinal sample, e.g., a fecal sample. Detecting the presence of levels of microorganisms that produce curli-related proteins such as sgA For example, proteins can be detected by ELISA, Western blot, lateral flow spectroscopy, and the like. It can be detected by immunoassays such as low-level assays and no-wash assays. For example, microorganisms that produce microbial proteins can be identified by nucleic acid analysis (qualitative or quantitative PCR, For example, microarray analysis or sequencing can be used to detect the Nucleic acids encoding physical proteins can be identified by qualitative or quantitative PCR, microarray analysis, etc. This can be detected by sequencing or branched DNA analysis. Presence of bacterial proteins or microorganisms in the intestine, or more bacterial proteins (if any) than in controls The level of the protein (or the microorganisms that produce the protein) determines whether the subject needs the composition. By way of example, suitable controls include bacterial proteins (or A subject negative for the genotype (microorganisms that produce genotype proteins), e.g., a healthy individual or a person in the intestine Individuals identified as not having bacterial proteins (or microorganisms that produce bacterial proteins) In some embodiments, the method includes the step of: the presence or level of curli (or curli-related proteins such as CsgA), or detects intestinal levels of microorganisms that produce intestinal curli-related proteins (e.g., CsgA) In some embodiments, the subject has an amyloid disorder and administers the composition. In some embodiments, the subject is identified as a member of a subpopulation of subjects in need thereof. The method further includes confirming the reduction or absence of intestinal amyloid aggregates after administration. In some embodiments, the method further comprises confirming a reduction or absence of intestinal amyloid protein after administration. The method further includes:

[0177] The compositions of the present disclosure, in some embodiments, inhibit alpha-synuclein aggregates (e.g., protozoa). The formation of amyloid fibrils, Lewy bodies, or other aggregates or other host amyloids occurs in the gastrointestinal tract. inhibits the aggregation of α-synuclein or other host amyloids at the point of initiation, thereby preventing can reduce microbial-induced amyloid aggregation, which may function as a mold or seed; It can also reduce α-synuclein in the digestive tract without crossing the blood-brain barrier. By targeting klein or other host amyloid aggregates, drugs can cross the blood-brain barrier. This eliminates the need for steroids, allowing for efficacy at lower doses and reducing systemic exposure to side effects Furthermore, it inhibits α-synuclein or other host amyloid aggregation, which it initiates. By targeting the disease at this time, it is possible to intervene early in the disease process and prevent motor symptoms or other neurological disorders. It becomes possible to prevent or inhibit disease progression before degenerative symptoms develop. In accordance with some embodiments of the compositions and methods described herein, alpha-synuclein aggregation in the gastrointestinal tract can be inhibited. By targeting, the gastrointestinal dysfunction is addressed and / or the gastrointestinal symptoms or behavior of the subject are improved. It can also improve bowel function, which can be seen in, for example, dysphagia, decreased intestinal motility, gastroparesis, and bowel obstruction. Constipation (including chronic constipation and chronic idiopathic constipation), small intestinal bacterial overgrowth (SIBO), diarrhea, abdominal pain and / or cramping, bloating, flatulence, nausea or any other symptoms of irritable bowel syndrome (IBS) , inflammatory bowel disease (IBD, e.g., ulcerative colitis and Crohn's disease), intestinal hyperpermeability, saliva Hypersecretion (excessive salivation), anorectal dysfunction, bowel dysfunction, or any of these The composition may include one or more of the following combinations:

[0178] Host amyloid aggregation in the brain as an approach to treat or inhibit neurodegenerative diseases In addition to targeting bacterial amyloid aggregation, targeting urinary tract infections (UTs) I) and other infectious diseases. In both cases, the present invention provides novel therapeutic approaches for the treatment of infectious diseases, such as The compounds tested are those that target the target of interest (e.g., brain for α-synuclein, UTI for It has been identified as having the ability to inhibit the amyloid aggregation process in tissues (urinary mucosa). do.

[0179] In some embodiments, the compositions and methods of the present disclosure target host alpha, such as alpha-synuclein. Inhibition of the interaction between amyloid and bacterial amyloids such as curli or adhesive pili In some embodiments, the present disclosure contemplates the use of the compounds described herein as therapeutic agents. The compositions and methods of the present invention target host amyloid in peripheral tissues such as the gastrointestinal tract or nasopharynx, but not in the brain. The compounds described herein can be used as inhibitors of amyloid aggregation and / or promoters of amyloid disaggregation. The compositions and methods of the present disclosure act locally in the gastrointestinal tract and are intended to be used in peripheral tissues. Modified derivatives of the compounds described herein that are essentially not absorbed into tissue, e.g., amyloid inhibitors Parenteral biological agents that retain their toxicity but do not cross the gastrointestinal epithelium or enter the primary circulation Further contemplated are derivatives available therefor.

[0180] In some embodiments, the compositions and methods of the present disclosure are directed to the lower small intestine, large intestine, and / or colon. The present invention contemplates a formulation that allows delivery of the composition to the site of action of the stomach or Enteric-coated tablets, capsules, and liquid gels are used to inhibit drug release in the upper GI tract. Alternatively, the composition may be contained in an inner enteric coated capsule or similar container. The capsule composition may contain a solid dosage form, which dissolves at or near the site of action. For example, preferably in the lower GI tract, more particularly in the lower small intestine. , EnTrinsic® intrinsic enteric capsules that dissolve in the large intestine or colon. In some embodiments, the composition is not absorbed and remains in the GI tract. do.

[0181] The compositions and methods of the present disclosure target one or more components of amyloidogenesis to inhibit gut-restrictive microorganisms. Molecular inhibitors are contemplated. Exemplary compounds of the invention include a polyphenol moiety. The compositions and methods of the present disclosure also include amylase inhibitors. Gut selection of a non-polyphenol class known to have the ability to inhibit eoloid formation The compositions and methods according to the present disclosure contemplate parenterally absorbable derivatives, such as those described above. Parenterally absorbable enteroselective derivatives or formulations of polyphenols or non-polyphenolic compounds are also included. Further plans.

[0182] As used herein, "subject" means any object that is not a target of interest and that has the same meaning as understood by those of skill in the art in light of this disclosure. and has its usual meaning. It is intended to describe neurological disorders or conditions associated with microbial-induced amyloidosis. Neurological disorders, such as Parkinson's disease, Lewy body disease, incidental Lewy body disease, and Alzheimer's disease Lewy body variant of Marr's disease, multiple system atrophy, pure autonomic failure, intestinal dysbiosis, Intestinal hyperpermeability, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), ulcerative colitis, Selected or specific for the diagnosis, treatment, inhibition, or amelioration of Rohn's disease, or any combination thereof. The term "animals" refers to animals such as, but not limited to, humans, dogs, cats, horses, donkeys, mules, and cattle. , domestic buffalo, camel, llama, alpaca, bison, yak, goat, sheep, pig, It refers to non-human mammals such as razor, deer, domestic antelope or non-human primates.

[0183] "Diagnosing" means any of the following: It has meaning when the subject has Parkinson's disease, dementia with Lewy bodies, incident Lewy body syndrome, Somatoform disorders, Lewy body variant of Alzheimer's disease, multiple system atrophy, pure autonomic failure, or or any combination thereof. May refer to the act or process of determining whether a person exhibits any symptoms or signs of a disorder. It also targets patients with intestinal dysbiosis, intestinal hyperpermeability, and irritable bowel syndrome (IBS). microbial-induced inflammatory bowel diseases such as inflammatory bowel disease (IBD), ulcerative colitis and / or Crohn's disease The act or process of determining whether a patient exhibits any symptoms or signs of gastrointestinal disorders related to steroids Diagnosing can also refer to the process of examining a subject's body or any of its tissues, fluids, components, It may further include determining whether the organ or part contains microbially-induced amyloid. Diagnosing means determining whether a subject's body, or any tissue, fluid, component, organ, or part thereof, Contains any factor that may affect the rate of aggregation or disaggregation of microbial-induced amyloid. The method may further include determining whether

[0184] "Subject suspected of having" means any of the conventional or non-standard substances that would be understood by one of ordinary skill in the art in light of this disclosure. It has its usual meaning: a subject who exhibits one or more clinical signs of a disease or condition. In certain embodiments, the disease or condition is Parkinson's disease, dementia with Lewy bodies, incident dementia, Lewy body disease, Lewy body variant of Alzheimer's disease, multiple system atrophy, pure autonomic nervous system In some embodiments, the present invention may include one or more of the following: These disorders include intestinal dysbiosis, intestinal hyperpermeability, irritable bowel syndrome (IBS), and inflammation. The disease may be selected from the group consisting of inflammatory bowel disease (IBD), ulcerative colitis or Crohn's disease.

[0185] "Subject in need thereof" means any of the conventional methods as would be understood by one of ordinary skill in the art in light of this disclosure. It has one common meaning: it requires the diagnosis of a disorder associated with amyloid formation. or a neurological disease or disorder associated with microbial-induced amyloid, such as Parkinson's disease Lewy body dementia, dementia with Lewy bodies, incidental Lewy body disease, Lewy body changes in Alzheimer's disease Treating or inhibiting atypical, multiple system atrophy, pure autonomic failure, or any combination thereof; In another embodiment, the present invention relates to a subject who has been selected or identified as being in need of improvement. Harmful effects include intestinal dysbiosis, increased intestinal permeability, irritable bowel syndrome (IBS), and inflammatory bowel disease ( IBD), ulcerative colitis or Crohn's disease.

[0186] "Bacteria" and "microorganism" (and the related terms "bacterial" and "microbial") are used herein. As used herein, the term "organisms" refers to organisms of the art that include bacteria, fungi, viruses, protists, archaea, etc. It is a recognized term in the field.

[0187] As used herein, "microbially-induced amyloid" refers to a compound that would be readily identified by one of skill in the art in light of the present disclosure. It has its customary and ordinary meaning as understood by those skilled in the art. It is a mammalian or microbial protein. amyloid fibrils produced by contact of proteins with one or more microbial proteins or The microbial protein aggregates refer to aggregates of one or more proteins of bacterial or fungal origin. However, the present disclosure is not limited to bacteriophages, viruses, bacteria, ancient Antibodies produced by interactions with proteins from bacteria, fungi, and other eukaryotes Contemplate myloid.

[0188] "Therapeutic effect" has its conventional and ordinary meaning as understood by those of skill in the art in light of this disclosure. It has the effect of alleviating to some extent one or more of the symptoms of the disease or disorder and curing the disease or disorder. "Cure" means the disappearance of symptoms of active disease. However, even after a cure is achieved, certain long-term or permanent effects of the disease may remain. (e.g. tissue damage).

[0189] "Improvement" shall have its conventional and ordinary meaning as understood by those of ordinary skill in the art in light of this disclosure. It refers to a decrease in the severity of at least one symptom of a condition or disease. In certain embodiments, improvement is a delay or slowing of the progression of one or more symptoms of the condition or disease. The severity of symptoms may be determined by subjective or objective measures known to those skilled in the art. It is possible.

[0190] "Regulate" has its conventional and ordinary meaning as understood by those of ordinary skill in the art in light of this disclosure. It refers to any factor in the body of a subject or any of its tissues, fluids, components, organs or It refers to a change in the presence, absolute level, relative level, function, or activity of a moiety. In certain embodiments, modulation refers to an increase in gene expression. In certain embodiments, modulation refers to an increase or decrease in the total serum level of a particular protein. In certain embodiments, modulation refers to an increase in the free serum levels of a particular protein or In certain embodiments, modulation refers to an increase or decrease in the aggregation state of a protein. In certain embodiments, the modulation involves increasing or decreasing the stability of amyloid fibrils. In certain embodiments, modulation refers to increasing the length, width, spacing, or density of amyloid fibrils. In certain embodiments, modulation refers to the modulation of specific non-protein factors, e.g. For example, it refers to an increase or decrease in the total serum level of a metabolite. In certain embodiments, the term "antibody" refers to an increase or decrease in the free serum levels of certain non-protein factors. The term refers to an increase or decrease in the total bioavailability of a particular protein. In some embodiments, modulation can be an increase or decrease in the total bioavailability of a particular non-protein factor. In certain embodiments, modulation refers to a change in the aggregation state of a protein. In some embodiments, modulation refers to a change in the rate or extent of aggregation or disaggregation of microbial-induced amyloid. vinegar.

[0191] In some compositions and methods of some embodiments according to the present disclosure, the subject is a In some embodiments, the patient is selected or identified to receive the composition described above. The subject is selected or identified as having high levels of Curli in the gastrointestinal tract. Such selection can be made by clinical or diagnostic evaluation. In this case, the subject is selected as having high levels of microbial-induced amyloid in the gastrointestinal tract. Such selection can also be made by clinical or diagnostic evaluation. In some embodiments, the subject has high levels of alpha-synuclein in the gastrointestinal tract. Similarly, such selection may be based on clinical or diagnostic evaluation. In some further embodiments, the subject has anosmia. , olfactory disturbance, bradykinesia, ataxia, tremor, muscle rigidity, posture and balance disorders, loss of motor activity aphasia, dysarthria or other changes in speech, changes in handwriting, orthostatic hypotension, memory impairment, swallowing problems, including abstinence, sleep disorders, cardiac arrhythmia, visual disturbances, depression and visual, auditory, olfactory or tactile hallucinations Psychiatric problems, dizziness, cognitive impairment, changes in dopamine levels, and serotonin levels changes, and / or changes in kynurenine levels, gastroparesis, anorectal dysfunction, and bowel dysfunction and (iii) a subject exhibiting one or more symptoms of a neurodegenerative disorder, such as a neurodegenerative disorder, a neurodegenerative disorder, or any combination thereof. In some embodiments, the subject is a neurologist in the field of diagnosis of neurological and amyloid disorders. The patient has been diagnosed with an amyloid disorder according to known methods. In embodiments, the subject is diagnosed with dementia with Lewy bodies, incident Lewy body disease, Alzheimer's disease, or Lewy body variant of leukemia, multiple system atrophy, pure autonomic failure, or any combination thereof In some embodiments, the subject has been diagnosed with or is at risk of having a combination of the above. The subject further exhibits gastrointestinal symptoms. In some further embodiments, the gastrointestinal symptoms include: Constipation, diarrhea, abdominal pain and / or cramps, bloating, flatulence, nausea or irritable bowel syndrome (IBS) Any other condition of inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, intestinal hyperpermeability or any other symptoms in any combination thereof.

[0192] In some embodiments, the compound or pharmaceutical composition is a compound or pharmaceutical composition for treating anosmia, dysosmia, bradykinesia, , ataxia, tremors, muscle rigidity, posture and balance disorders, loss of motility, dysarthria or other Changes in speech, changes in handwriting, orthostatic hypotension, memory loss, swallowing problems, incontinence, sleep disorders, cardiac arrhythmias psychiatric problems including palpitations, visual disturbances, depression and visual, auditory, olfactory or tactile hallucinations; cognitive impairment, changes in dopamine levels, changes in serotonin levels, changes in kynurenine levels before the appearance of neurological symptoms or conditions, such as changes in It is administered.

[0193] In the compositions and methods according to some embodiments of the present disclosure, a subject selected for treatment In some embodiments, the subject selected for treatment is: The subject may be between 17 and 30 years old. In some embodiments, the subject selected for treatment is: In some embodiments, the subject selected for treatment is: In some embodiments, the subject selected for treatment is: In some embodiments, the compositions and methods described herein may be used in combination with a steroid or anti-inflammatory drug. Subjects selected for treatment by the method may be over 69 years of age.

[0194] In compositions and methods according to some embodiments of the present disclosure, the compositions described herein administration of one or more of the following to treat amyloid or microbial Prevent the formation of induced amyloid or detoxify amyloid or microbial-induced amyloid. It has a promoting effect on aggregation and does not need to cross the blood-brain barrier, for example. In embodiments, administration of one or more of the compositions described herein is to the gastrointestinal tract, nasal cavity, olfactory bulb or inhibits further aggregation of amyloid or microbial-induced amyloid in enteric neural tissues In some embodiments, the effect is achieved without the need to cross the blood-brain barrier, for example. Administration of one or more of the compositions described herein may result in a steroid effect in the gastrointestinal tract, nasal cavity, olfactory bulb, or enteric nervous tissue. The effect of causing or promoting disaggregation of amyloid or microbial-induced amyloid in In some embodiments, the present invention provides a method for the production of steroid hormones, e.g., without the need to cross the blood-brain barrier. Administration of one or more of the compositions described herein may result in a decrease in the level of inflammatory bowel disease in the gastrointestinal tract, nasal cavity, olfactory bulb, or enteric nervous system tissue. The effect of causing or enhancing disaggregation of existing amyloid or microbial-induced amyloid In some embodiments, the present invention provides a method for the production of steroid hormones, e.g., without the need to cross the blood-brain barrier. Administration of one or more of the compositions described herein may be used to treat one or more neurological or neurodegenerative disorders. In some embodiments, the compounds described herein are effective in preventing the onset of the above symptoms. Administration of one or more of the compositions of the present invention may be used to treat one or more symptoms of one or more neurological or neurodegenerative disorders. In some embodiments, one of the compositions described herein provides an effect of improving the condition of the skin. The administration of the above has the effect of reversing one or more symptoms of one or more neurological or neurodegenerative disorders. In some embodiments, the one or more symptoms of the one or more neurological disorders include anosmia. anosmia, olfactory disturbance, bradykinesia, ataxia, tremor, muscle rigidity, posture and balance disorders, motility loss of speech, dysarthria or other changes in speech, changes in handwriting, orthostatic hypotension, memory problems, swallowing problems , incontinence, sleep disorders, cardiac arrhythmias, visual disturbances, depression and visual, auditory, olfactory or tactile hallucinations. Psychiatric problems, including dizziness, cognitive impairment, changes in dopamine levels, and serotonin levels changes in blood cholesterol and / or kynurenine levels, gastroparesis, anorectal dysfunction, bowel coordination In some embodiments, the present invention includes one or more of: Alternatively, the one or more neurological disorders may include an amyloid disorder. In some cases, one or more of the above neurological disorders may be dementia with Lewy bodies, incident Lewy body disease, or atopic dermatitis. Lewy body variant of Ilzheimer's disease, multiple system atrophy, and / or pure autonomic failure, or It may include one or more of any combination of these.

[0195] In another embodiment, the inhibitor of amyloid formation is administered systemically or locally to the gut of the central nervous system. For example, mammalian amyloid or mammalian amyloid precursor Effective inhibitors of the aggregation of somatic proteins are also useful for the amyloid disorders described herein (Table 2). Thus, in such embodiments, amyloid may be useful in the treatment of one or more of the following: The compositions containing the inhibitors of formation can be administered systemically (e.g., intravenously, subcutaneously, intramuscularly, intraperitoneally) or intraperitoneally. includes local administration (e.g., local injection near the vagus nerve, intraspinal injection, or intracranial injection) It can be formulated for parenteral administration. To be delivered to the CNS, the inhibitor must cross the blood-brain barrier. Therefore, in such an embodiment, the inhibitor is preferably a lipid It is a water-soluble molecule, or can be modified to increase lipid solubility, or can cross the blood-brain barrier. The compound may be co-administered with a compound that enhances perfusion (see, for example, WO 20140766 55, International Publication No. 2012159052, and International Publication No. 1992018529 (I want to be).

[0196] In some embodiments of the present disclosure, compositions and methods may be used to treat a variety of conditions, including: The levels of microbial amyloid and / or microbial-induced amyloid are monitored during the course of treatment. In some further embodiments, amyloid and / or microbial-induced The level of conductive amyloid is monitored before and / or after a course of treatment. In some embodiments, the level of alpha-synuclein in the subject's tissue, body fluid, or feces is measured over the course of treatment. In some embodiments, the level of alpha-synuclein is monitored at In some embodiments, amyloid is monitored before and / or after a course of treatment. Measurements of microbial-induced amyloid and / or alpha-synuclein were obtained from the subject. In some embodiments, amyloid, microbial-induced amyloid, Measurements of α-synuclein and / or α-synuclein are measured in tissue samples obtained from the subject. In some embodiments, the tissue sample is selected from the group consisting of intestinal epithelium, peritoneum, enteric nerve tissue, olfactory tissue, and the like. In some embodiments, the tissue includes the nasal lining, the sinus lining, the brain, and / or neural tissue. The tissue sample comprises cerebrospinal fluid or synovial fluid. In some embodiments, the tissue sample comprises blood , lymph or plasma.

[0197] Methods for identifying compounds Bacterial amyloid causes aggregation and amyloid formation of the eukaryotic protein α-synuclein Disclosed herein are methods for identifying compounds that alter the ability of a compound to promote neurodegenerative disease. Methods for screening for entities useful in treating or inhibiting neurodegenerative diseases, and for predicting the progression of neurodegenerative diseases Further disclosed herein are methods for screening for entities useful in the prevention or amelioration of neurological disorders. Methods for screening entities useful in treating or inhibiting gastrointestinal dysfunction associated with degenerative diseases Further disclosed herein are the molecular pathogenesis of mammalian amyloid diseases and bacterial amyloidosis. Methods for studying the molecular link between myloidogenesis and mammalian amyloidogenesis are disclosed. According to the methods of the present disclosure, the neurodegenerative disease and / or mammalian Physical amyloid diseases include Parkinson's disease (PD), dementia with Lewy bodies, multiple system atrophy, Multiple sclerosis (MS), frontotemporal dementia (FTD), REM sleep behavior disorder (RBD), α -Synucleinopathy, PD-related constipation, PD-related hypotension, Huntington's disease, Alexandria Dart's disease, amyotrophic lateral sclerosis (ALS), or Alzheimer's disease, and amyloid-related These may include other diseases that cause

[0198] The methods disclosed herein include (1) detecting bacterial amiami on the bacterial surface or in the proximal extracellular space; (2) in the gastrointestinal tract or olfactory system (including enteroendocrine cells and enteric neurons) Interaction of bacterial amyloid with α-synuclein, or (3) the gastrointestinal tract (enteroendocrine cells and and aggregation of α-synuclein in enteric neurons (including enterocytes). According to the methods of the present disclosure, alpha-synuclein can be detected in a broader range of in vitro assays. Representative of known mammalian amyloids or mammalian amyloid precursor proteins should be considered an amyloid protein, and the methods disclosed herein involve the use of one amyloid protein. Amyloid proteins (bacterial or human) cause aggregation of other amyloid proteins, It can be adapted by one skilled in the art to assess aggregation of any amyloid protein. Representative disorders exhibiting idiopathogenesis and proteins involved in these disorders are described in the present disclosure. The methods can be evaluated using methods including, but not limited to, those disclosed in Table 2. Thus, in some embodiments, the method comprises administering a plurality of concentrations of the compound in the presence of the compound. Microbial amyloid or microbial amyloid precursors were incubated with multiple concentrations of α-synuclein and and / or other mammalian amyloids or mammalian amyloid precursors, analyzing or measuring the formation or disaggregation of amyloid after the reaction; and and comparing the measurement to a control assay or measurement, said control being the absence or presence of said composition. The method disclosed herein includes analyzing or measuring the formation of amyloid after the above reaction under In certain methods and compositions, the microbial amyloid or microbial amyloid precursor is contains CsgA.

[0199] In some embodiments, the methods according to the present disclosure include: Alternatively, a microbial amyloid precursor (e.g., a composition containing CsgA) may be administered to mammalian cells at various concentrations. The composition is intended to be contacted with a physical amyloid or a mammalian amyloid precursor, tested for its ability to inhibit amyloid formation or enhance amyloid disaggregation. In some further embodiments, the compound or mixture is a microbial amyloid. or microbial amyloid precursors, mammalian amyloids or mammalian amyloid precursors and the above combination of test compositions are analyzed or tested for changes in the amount of amyloid present. In some further embodiments, microbial amyloid or microbial amyloid is measured. amyloid precursor, mammalian amyloid or mammalian amyloid precursor, and a test composition The rate and / or extent of amyloid formation in the combination is compared to a control lacking the composition. The rate of amyloid formation in the sample is compared to the rate of amyloid formation in the sample. In some further embodiments, the total amount of amyloid formation is measured. In some further embodiments, the temperature of the assay is changed, thereby The stability of newly formed amyloid fibrils was compared with those formed under native conditions. In some embodiments, the method comprises measuring the composition in a multi-well assay plate. In some further embodiments, the method is performed by placing the plate in a well. The method according to the present disclosure is carried out in the presence of a physical agitator. The physical agitators include glass, Teflon, or polymer beads. In some further embodiments, the polymer beads are made of polystyrene, polylactic acid, poly Polylactic-co-glycolic acid, polycarbonate, or polytetrafluoroethylene (Te In some embodiments, the beads used for agitation may include Freon® beads. In some embodiments, the diameter of the beads or objects is between 10 and 1000 μm. The beads or objects used for stirring have a longest dimension of 10 to 100 μm, 80 to 200, 180~300μm, 280~400μm, 380~500μm, 480~600μm, 580~700μm, 680~800μm, 780~900μm, or 880~1000 In some embodiments, the beads or objects used for agitation are at their finest. In some embodiments, the beads or objects used for agitation have a major dimension greater than 1 mm. In certain embodiments, the beads or objects are , 1~3mm, 1~5mm, 2~5mm, 3~5mm, 4~5mm, 5~6mm, 5~7 mm, 5~8mm, 5~9mm, 5~10mm, 2~10mm, 4~10mm, 6~10 In certain embodiments, the beads or objects are The longest dimension of these is 1mm, 2mm, 3mm, 4mm, 5mm, 6mm, 7mm, 8mm, 9mm mm, or 10 mm.

[0200] In some embodiments, the microbial amyloid or microbial amyloid precursor is an adhesion CsgA, the major protein component of curli, also known as sex pili, or its related In some embodiments, microbial amyloid or The microbial amyloid precursor is a precursor to the conversion of CsgA to its amyloid form or to a precursor derived from it. In some embodiments, the mammalian animal comprises CsgB, which is the core polypeptide that binds to the mammalian animal. Myloid or mammalian amyloid precursors include α-synuclein.

[0201] In some embodiments, microbial amyloid or microbial amyloid precursors (e.g., , CsgA) at various concentrations of mammalian amyloid or mammalian amyloid The contacting of the amyloid precursor with the composition is carried out in the presence of an indicator of amyloid formation. In some further embodiments, the indicator may include a fluorescent indicator, and the fluorescent intensity of the indicator The degree of amyloid accumulation varies in a manner that correlates with the amount of amyloid present in the sample. The fluorescence changes associated with the changes in the molecular environment associated with the incorporation of the label into the amyloid fibrils. In some further embodiments, the indicator is thioflavin T (T In some embodiments, the label attached to the amyloid precursor molecule may comprise: Emission by intermolecular fluorescence quenching or fluorescence resonance energy transfer correlated with the formation of amyloid fibrils. They may exhibit a change in the intensity or wavelength of light. Exemplary fluorescent labels are listed in the Molecular Probes Handbook ( Invitrogen, Inc., 2010), which describes a FRET pair, For its teachings on fluorescence quenching and fluorescent probes that can be conjugated to proteins Other exemplary fluorescent labels include, but are not limited to: However, green fluorescent protein (GFP), yellow fluorescent protein (YFP), AmCyan1, AsREd2, mBanana, mCherry, Dendra2, DsRed2, Ds Red-express, DsRed-monomer, DsRed, E2-Crimson, GFP-UV, blue fluorescent protein (BFP), HcRed1, mOrange, PAm Cherry, mPlum, mRaspberry, mStrawberry, tdTo mato, ZsGreen1, ZsYellow1, or AcGFP1, or any of them or other fluorescent proteins known in the art. In some further embodiments, the label attached to the mammalian amyloid precursor is This is different from the label attached to bacterial amyloid or bacterial amyloid precursor. In this form, bacterial amyloid or bacterial amyloid precursors are not labeled. In embodiments, mammalian amyloid, mammalian amyloid precursor, bacterial amyloid The amyloid precursor or bacterial amyloid comprises two or more labels. The indicator may be a colorimetric indicator, a spin label (e.g., 3H, 15N, or 13C), , metal ion binding compounds (e.g., porphyrins, chelators, polyhistidine, or other metal-binding polypeptides), enzymes, or amyloid-specific antibodies. In some embodiments, the development of amyloid fibrils is observed directly by light microscopy. In some embodiments, amyloid formation is observed by direct light transmission or by reflectance. In some embodiments, amyloid formation is observed by total internal reflection FTIR. In some embodiments, amyloid formation is observed by NMR, FTIR, SPIR or is observed by SPR spectroscopy. In some embodiments, the sample is visualized and / or confirmed by birefringence. In some embodiments, amyloid formation is monitored by Raman spectroscopy. In some embodiments, amyloid formation is observed by scattering of the intrinsic fluorescence of the sample. Changes in, for example, internal tryptophan, tyrosine, phenylalanine, histidine, and This is observed by monitoring the changes caused by arginine residues. In embodiments, amyloid formation is determined by binding of an amyloid-specific antibody, as described in the art. By known means, for example, fluorescent labels, colorimetric labels, spin labels, radioisotopes, and by conjugation of the antibodies to enzymes, fluorescent proteins, and metal-binding domains. or by other methods known to those skilled in the art for the detection or visualization of antibodies. According to the methods described herein, the antibody is It may include antibodies that have selective binding activity to either amyloid or amyloid precursors. .

[0202] In some embodiments, the methods of the present disclosure comprise: The dynamics of the fluorescence intensity of amyloid-specific dyes in the presence of bacterial amyloid precursors or aggregates. In some embodiments, the mammalian state may be monitored. The physical amyloid precursor is alpha-synuclein. The amyloid precursor or aggregate is CsgA. In some embodiments, the amyloid The do-specific dye is thioflavin T.

[0203] In some embodiments, the present disclosure provides kits for carrying out the methods described herein. In some embodiments, the kit comprises at least a mammalian amyloid or mammalian amyloid precursor, bacterial amyloid or bacterial amyloid precursor, and an indicator of amyloid formation as described herein, such indicator comprising a mammalian amyloid. mammalian amyloid precursors, bacterial amyloid precursors, and and one or more reaction vessels, which may or may not be conjugated to the above. The kit may include a multi-well plate. The kit may be used to carry out the methods described herein. It may further include instructions for administration.

[0204] The methods of the present disclosure involve the aggregation and / or disaggregation of amyloid, particularly microbially induced amyloid. Methods for screening candidate compounds to identify compounds that modulate the expression of some In some embodiments, the methods of the disclosure involve screening a library of candidate compounds. In some further embodiments, mammalian animals are cultured according to the methods disclosed herein. amyloid or mammalian amyloid precursor and bacterial amyloid precursor or bacterial amyloid The composition contacting the amyloid inhibits amyloid formation or destabilizes existing amyloid. One or more compounds or combinations thereof suspected in the art to stabilize or disaggregate In certain embodiments, mammalian animals are treated according to the methods disclosed herein. amyloid or mammalian amyloid precursor and bacterial amyloid precursor or bacterial amyloid The composition that is contacted with the id comprises a natural product or an extract from a natural product. In some embodiments, mammalian amyloid or mammalian amyloids can be prepared according to the methods disclosed herein. and a composition for contacting the bacterial amyloid precursor or bacterial amyloid, In some embodiments, the composition comprises: an herb, herbal extract, or botanical. It may include tissues or fluids obtained from animals, plants, or fungi. In some embodiments, the composition may be applied to the seeds, fruits, flowers, leaves, stems, cambium, or roots of a plant, or to any of its components. The composition may include tissues, fluids, or tissue or fluid extracts obtained from a combination of these. In some further embodiments, the composition is obtained by extracting feces, urine, blood, lymph, synovial fluid, or the like from the animal. cerebrospinal fluid, saliva, or tissue, fluid, or tissue or fluid extract obtained from any internal organ In some embodiments, the composition may comprise one or more bacteria or lysates. , extracts, conditioned culture media, freeze-dried bacteria, freeze-dried lysates, freeze-dried culture media thereof, or In some embodiments, the bacterium is a bacterium selected from the group consisting of bacterium, bacterium thuringiensis ... Bacteroides, Prevotella, Parabacterium Parabacteroides, Faecalibacterium ibacterium), Eubacterium, Roseburia ( Roseburia, Blautia, Coprococcus ccus and Bifidobacterium, or Any combination of one or more of the following may be included:

[0205] In some embodiments, the methods of the present disclosure are used to treat a subject with an amyloid disorder. The disclosed method can be used to diagnose or assess the risk of Parkinson's disease, Lewy body syndrome, and the like. Alzheimer's disease, incidental Lewy body disease, Lewy body variant of Alzheimer's disease, multiple system atrophy Treatment of one or more neurological disorders, including cerebrovascular disease, pure autonomic failure, or any combination thereof The above disorders include tremors, paralysis, dyskinesia, and the like. Neurological disorders such as diarrhoea, and / or constipation, diarrhea, abdominal pain and / or cramps, bloating, flatulence Intestinal, nausea or any other symptoms of irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), ulcers one or more of ulcerative colitis, Crohn's disease, intestinal hyperpermeability, or any combination thereof In addition to the physical symptoms known in the field of diagnosis and treatment of gastrointestinal symptoms such as those listed above, Clinical presentation of neurological disorders such as nication symptoms, stereotypic behaviors, sensorimotor problems, and / or anxiety-like behaviors These may include behavioral symptoms as known in the field of clinical diagnosis and treatment. Such clinical and / or diagnostic evaluations and determinations may be used to perform one or more of the methods provided in this disclosure. and / or identifying one or more subjects receiving one or more compounds described herein according to The methods of the present disclosure may, in some embodiments, be used to diagnose neurological disorders. and monitoring behavioral, physical, and / or gastrointestinal symptoms as known in the art of steroid therapy. In some embodiments, methods according to the present disclosure may include detecting changes in a subject's behavior. In some further embodiments, the method according to the present disclosure incorporates monitoring of the catalysis. Parkinson's disease, dementia with Lewy bodies, incident Lewy body disease, and Alzheimer's disease Related to Bee body variant, multiple system atrophy, pure autonomic failure, or any combination thereof Incorporate monitoring of subjects for behavioral symptoms known to be associated with In some further embodiments, the methods according to the present disclosure provide for the treatment of symptoms such as bradykinesia, ataxia, tremor, muscle rigidity, and the like. Shrinkage, posture and balance disorders, loss of motor activity, dysarthria or other changes in speech, changes in handwriting , orthostatic hypotension, memory impairment, swallowing disorders, incontinence, sleep disorders, cardiac arrhythmias, visual disturbances, depression and psychiatric problems, including visual, auditory, olfactory, or tactile hallucinations, dizziness, cognitive impairment, or Any combination of these, or amyloid disorders, especially α-synucleinopathies Any other condition known to those in the field of neurological diagnosis or treatment that is useful in diagnosis. In some further embodiments, the present disclosure incorporates monitoring of the subject. The method is aimed at treating bowel movements including gastroparesis, colonic motility, anorectal dysfunction, and bowel dyssynergia. Incorporate monitoring of subjects for activity. Again, such clinical outcomes and / or diagnostic evaluations and determinations according to the methods described herein. can identify and / or select one or more subjects for treatment. In some embodiments, the disclosed methods may include, in addition to the clinical monitoring described above, the use of the cell Monitoring levels of fungal amyloid, host-derived amyloid, and microbial-induced amyloid According to the methods of the present disclosure, the amyloid may be collected from the subject's digestive tract, feces, urine, The method of the present disclosure may be monitored in blood, saliva, cerebrospinal fluid, and / or synovial fluid. Monitoring of said amyloid in any tissue or body fluid obtainable from the subject during the course of treatment. The present invention contemplates the use of amyloid-inducing agents in a sample containing amyloid-inducing agents, thereby determining whether the sample contains a factor that promotes or inhibits amyloid formation. In some embodiments, the assays described herein determine whether amyloid forms the presence of factors that enhance or accelerate the development of the target tissue, body fluid, or other sample from which it is derived. Elephants may be considered at high risk for developing amyloid disorders. In this case, the subject is administered a drug or treatment to ameliorate or prevent the amyloid disorder. Again, such clinical and / or diagnostic evaluations and determinations may be used to: According to one or more of the methods provided in this disclosure, one or more of the compounds described herein may be One or more subjects can be identified and / or selected to receive the treatment.

[0206] According to the methods disclosed herein, treating or inhibiting a disorder involving amyloid formation includes administering to a subject a therapeutically effective amount of a compound selected from the group consisting of steroids, steroid hormone ... and steroid hormones. This can be achieved by adjusting the dosing schedule for administration of the composition, so that After experiencing an intermittent, partial, or complete reduction in medication over a period of time, the subject is given In some embodiments, the dose is administered daily for 1 to 30 days. In some embodiments, the drug-free period is 1 to 30 days. In some further embodiments, the compositions of the present disclosure are administered in the following manner: In some other embodiments, the drug may be completely cleared from the subject's body before administration of the drug. In some further embodiments, a dose less than the usual daily dose is administered throughout the period. It is possible that sub-therapeutically effective amounts of the administered composition remain in the subject during a drug holiday period. In some further embodiments, the administered composition is maintained at a therapeutic level in the affected tissue. In some embodiments, the composition is capable of remaining in the subject in a sufficient amount to sustain the intended effect. The subject is a patient with amyloidosis at any time after the onset of one or more of the aforementioned symptoms of a neurological disorder associated with amyloid formation. In some embodiments, the compositions according to the methods described herein are administered in one or more In some embodiments, the compound described herein is administered prior to the onset of symptoms of the above disorders. The compositions according to the methods described above are administered simultaneously with or after the onset of symptoms of one or more of the above disorders. It is given.

[0207] The following items are described in accordance with some embodiments of the present disclosure:

[0208] 1. Contacting amyloid or amyloid precursors with a composition comprising a compound described herein and / or inhibiting the formation of amyloid protein aggregates, comprising: .

[0209] 2. Contacting amyloid or amyloid precursors with a composition comprising a compound described herein A method for inhibiting the formation of amyloid protein aggregates, comprising:

[0210] 3. A method of disrupting the formation of amyloid protein aggregates in a subject, comprising: administering to said subject a composition comprising a compound described herein; and Optionally, prior to administering the composition, amylolytic activity may be assessed, such as by clinical or diagnostic evaluation. selecting said subject to benefit from a molecule that disrupts the formation of id aggregates; and / or teeth, Optionally, preventing or preventing the formation of amyloid aggregates in said subject after administration of said composition. comprising measuring the inhibition.

[0211] 4. Contacting amyloid or amyloid precursors with a composition comprising a compound described herein 20. A method for disrupting the formation of amyloid protein aggregates, comprising:

[0212] 5. Inhibit, improve, reduce the likelihood, delay the onset and treatment of amyloid disorders The present invention relates to a method for treating or preventing a disease, comprising administering a compound described herein or a pharmaceutical composition thereof to a patient in need thereof. administering to a subject in need thereof.

[0213] 6. Amyloid disorders include alpha-synucleinopathy, Parkinson's disease, and dementia with Lewy bodies. Alzheimer's disease, incidental Lewy body disease, Lewy body variant of Alzheimer's disease, multiple system atrophy, young adult or pure autonomic failure, or any combination thereof. 5. The method according to claim 5.

[0214] 7. The amyloid disorder is characterized by the presence of an intestinal amyloid protein or aggregate, such as CsgA. 7. The method according to any one of claims 5 to 6, comprising a protein or aggregate containing a bacterial protein. How to do it.

[0215] 8. Amyloid disorders are associated with intestinal dysbiosis, increased intestinal permeability, and irritable bowel syndrome (IBS) ), inflammatory bowel disease (IBD), ulcerative colitis, or Crohn's disease, any of items 5 to 7 1. The method according to claim 1.

[0216] 9. Bacterial proteins, microbial proteins such as CsgA, in the intestinal samples of the subjects or detecting the presence or level of a nucleic acid encoding a bacterial protein The method according to any one of items 5 to 8, further comprising:

[0217] 10. Intestinal samples, bacterial proteins or microorganisms that produce bacterial proteins are detected. If the presence of bacterial proteins or bacterial proteins is detected in the intestinal sample, If the level of the microorganism produced is greater than a predetermined level or a control, the subject is deemed to require the composition. 10. The method of claim 9, wherein the method is selected as essential.

[0218] 11. Further comprising confirming the reduction or absence of intestinal amyloid protein after administration. 2. The method according to any one of items 7 to 10.

[0219] 12. The method of any one of paragraphs 7 to 11, further comprising identifying the subject as having gastrointestinal symptoms. 10. The method according to any one of claims 1 to 9.

[0220] 14. Subjects with dysphagia, decreased intestinal motility, gastroparesis, constipation (chronic constipation and chronic idiopathic constipation) Constipation (including idiopathic constipation), small intestinal bacterial overgrowth (SIBO), diarrhea, abdominal pain and / or cramps, bloating Fullness, flatulence, excessive salivation (hypersalivation), anorectal dysfunction, bowel coordination disorder, and other adverse reactions 13. The method according to any one of claims 5 to 12, wherein the patient is suffering from a gastrointestinal symptom including one or more of the following: method.

[0221] 16. The method of claim 1, wherein the composition is formulated for enteral, oral, or intranasal delivery. 10. The method according to any one of claims 1 to 9.

[0222] 17. The composition is formulated for controlled release in the lower intestine or colon, 7. The method of any one of clauses 6.

[0223] 18. The composition is an enteric coated capsule, tablet, soft gel, spray-dried powder, or , polymer matrices, hydrogels, enteric coated solids, crystalline solids, amorphous solids, glassy solids, coated particles, liquids, sprayed liquids, aerosols or microspheres Item 18. The method according to any one of items 1 to 17, wherein the composition is a microcapsule.

[0224] 19. The amyloid protein is one or more mammalian proteins, e.g., α- Synuclein, tau, beta-amyloid derived from amyloid precursor protein, medin, amyloid Polypoprotein AI, atrial natriuretic factor, beta-amyloid, cystatin I APP (amylin), beta-2 microglobulin, transthyretin, PrP, gel Zolin, Lysozyme, Huntingtin, Keratoepithelin n), calcitonin, prolactin, serum amyloid A, SOD1 and / or immunoglobulins 19. The method according to any one of paragraphs 1 to 18, comprising any one or more of the following light chains: method.

[0225] 20. The amyloid protein is one or more bacterial or fungal proteins. 20. The method of any one of claims 1 to 19, comprising:

[0226] 21. The amyloid protein is a first to fourth amyloid protein, including a bacterial protein such as CsgA. 21. The method of any one of clauses 20.

[0227] 22. The amyloid protein is present in the gastrointestinal tract, dural sinuses, oral cavity, or nasal cavity. The method according to any one of items 1 to 21,

[0228] 23. The method according to any one of claims 1 to 22, wherein the amyloid protein is present in enteric nerve tissue or the olfactory bulb. 10. The method according to any one of the preceding claims.

[0229] 24. The method of any one of paragraphs 1 to 23, wherein the composition is administered daily.

[0230] 25. The method of any one of paragraphs 1 to 24, wherein the composition is administered multiple times per day. .

[0231] 26. The method of any one of paragraphs 1 to 25, wherein the composition is administered less frequently than daily. The method described.

[0232] 27. The composition is administered every 2, 3, 4, 5, 6, or 7 days. 27. The method of any one of items 1 to 24 or 26, wherein the method is administered to both the patient and the subject.

[0233] 28. During the administration process, enteric amyloid levels and / or amyloid protein 28. The method of any one of items 1 to 27, further comprising measuring or assessing:

[0234] 29. After the administration course, enteric amyloid levels and / or amyloid protein 29. The method of any one of items 1 to 28, further comprising measuring or evaluating.

[0235] 30. Measuring or evaluating changes in the nervous system, such as neurological symptoms or characteristics of a subject. 30. The method according to any one of items 1 to 29, comprising:

[0236] 31. The above subjects are under 18 years old, 18-30 years old, 30-50 years old, 50-60 years old, 60+ years old 31. The method of any one of paragraphs 1 to 30, wherein the patient is 70 years of age or over 70 years of age.

[0237] 32. Further measuring or evaluating changes in the gastrointestinal system, such as gastrointestinal symptoms or characteristics of a subject. 32. The method of any one of items 1 to 31, comprising:

[0238] 33. The method of claim 32, wherein the gastrointestinal symptoms include constipation.

[0239] 34. The subject has constipation, diarrhea, abdominal pain and / or cramps, bloating, flatulence, nausea, or irritability. any other symptoms of inflammatory bowel syndrome (IBS), inflammatory bowel disease (IBD), e.g., ulcerative colitis any other condition of the liver, kidney, liver cirrhosis, liver cirrhosis, liver cirrhosis, liver ulcers ... 34. The method of any one of clauses 1 to 33, wherein the patient is suffering from gastrointestinal symptoms including one or more of:

[0240] 35. The method of any of paragraphs 1 to 34, wherein the composition is administered after the onset of a neurological symptom or condition. The method according to any one of claims 1 to 4.

[0241] 36. The above neurological symptoms or conditions include anosmia, dysosmia, bradykinesia, ataxia, and tremor. , muscle rigidity, posture and balance disorders, loss of automatic movements, dysarthria or other changes in speech, handwriting changes in blood pressure, orthostatic hypotension, memory problems, swallowing problems, incontinence, sleep problems, cardiac arrhythmias, visual disturbances, cavities Depression and psychiatric problems including visual, auditory, olfactory or tactile hallucinations, dizziness, cognitive impairment , changes in dopamine levels, changes in serotonin levels, changes in kynurenine levels, and / or or any combination thereof.

[0242] 37. The method of any of paragraphs 1 to 36, wherein the composition is administered before the onset of a neurological symptom or condition. The method according to any one of claims 1 to 4.

[0243] 38. The method of any one of paragraphs 1 to 37, wherein the method is repeated.

[0244] 39. For a given administration, the composition is different from the previously administered composition, first to third 9. The method of any one of clauses 8.

[0245] 40. For a given administration, the dose administered is different from the previously administered dose, Item 39. The method according to any one of items 1 to 39.

[0246] 41. The composition contains caffeine, nicotine, theophylline, theobromine, xanthine 41. The method according to any one of claims 1 to 40, wherein the method is co-administered with a methylxanthine, a methylxanthine, or a derivative thereof. The method described in paragraph .

[0247] 42. Methods 1 to 4, further comprising administering an α-synuclein aggregation inhibitor to the subject. 2. The method according to any one of paragraphs 1 to 11.

[0248] 43. The subject is at risk of developing Parkinson's disease, as determined by clinical or diagnostic assessment. Any of paragraphs 1 to 42, which are identified or selected as having or already having a risk 1. The method according to claim 1.

[0249] 44. The subject is diagnosed, for example, by clinical or diagnostic assessment, with dementia with Lewy bodies, X-linked Lewy body disease, Lewy body variant of Alzheimer's disease, multiple system atrophy, pure autonomic nervous system as being at risk of developing or already having neurological disorders, or any combination thereof 44. The method of any one of paragraphs 1 to 43, wherein the antibody is identified or selected.

[0250] 45. Gastrointestinal symptoms are associated with Parkinson's disease or parkinsonism, as described in paragraph 14. How to do it.

[0251] 46. ​​Diagnosing an amyloid disorder by detecting the presence or level of enterobacterial amyloid protein. Item 45. The method according to any one of items 1 to 44, wherein the diagnosis can be made by measuring the level of the disease.

[0252] Additional options The following options are described in accordance with some embodiments of the present disclosure:

[0253] 1. A method for identifying a composition that affects the formation of microbially induced amyloid, comprising: (a) detecting multiple concentrations of microbial amylolytic enzymes in the presence of a composition comprising a compound described herein; Inject α-synuclein into cells containing α-synuclein or microbial amyloid precursors (e.g., CsgA) at multiple concentrations. contacting the (b) analyzing or measuring the formation of amyloid produced by the reaction described in (a); and (c) Comparing the analysis or measurement made under (b) with a control analysis or measurement. The control determines the formation of amyloid following the reaction described in (a) in the absence of the composition. The method includes analyzing or measuring, comparing.

[0254] 2. The microbial amyloid or microbial amyloid precursor comprises CsgA. The method described in 1.

[0255] 3. The method of option 1 or 2, further comprising stirring during (a).

[0256] 4. The contacting performed in (a) is performed in the presence of an indicator of amyloid formation. , the method described in options 1 to 3.

[0257] 5. The indicator is a fluorescent indicator, a spin-label indicator, an enzyme, an antibody, or a colorimetric indicator. The method described in option 4.

[0258] 6. The method of option 4, wherein the indicator is thioflavin T.

[0259] 7. The antibody has specificity for aggregated alpha-synuclein, and the antibody is optionally Optionally, a fluorescent label, an enzyme, a colorimetric label, a spin label, a metal ion binding moiety, a nucleic acid, a polysaccharide or The method of option 4, wherein the polypeptide is conjugated to the

[0260] 8. The CsgA and the α-synuclein are each separately labeled. 8. The method according to any one of 1 to 7.

[0261] 9. The formation of the above is intrinsic fluorescence, fluorescence of a dye or label, fluorescence resonance energy transfer, fluorescence Polarized light, Fluorescence polarization transfer, UV / Vis spectroscopy, Magnetic resonance, Raman scattering, Electron paramagnetic spin resonance by acoustic, optical, electron, scanning tunneling, or atomic force microscopy. The method according to any one of options 1 to 8, wherein the analysis or measurement is performed by

[0262] 10. The method of any of options 1-9, wherein the composition comprises a mixture of compounds.

[0263] 11. Any of options 1-10, wherein the composition comprises tissue, body fluid, or an extract thereof. The method described.

[0264] 12. The composition contains feces, urine, blood, cerebrospinal fluid, or saliva, or components thereof. A method according to any one of options 1 to 11.

[0265] 16. The composition comprises one or more bacteria, bacterial extracts, lysates, conditioned culture media, frozen culture media, or the like. Contains dried bacteria, lyophilized lysate, lyophilized culture medium, or any combination thereof. A method according to any of options 1 to 10.

[0266] 17. The method of claim 1, further comprising identifying or selecting a composition that alters amyloid formation. The method according to any one of claims 1 to 16.

[0267] 18. The method of claim 1, further comprising identifying or selecting a composition that reduces amyloid formation. The method according to any one of claims 1 to 17.

[0268] 19. In (b), the rate of amyloid formation is analyzed or measured. A method according to any one of the preceding claims.

[0269] 20. A method for producing microbially-induced amyloid, comprising: (a) detecting multiple concentrations of microbial amylolytic enzymes in the presence of a composition comprising a compound described herein; Inject α-synuclein into cells containing α-synuclein or microbial amyloid precursors (e.g., CsgA) at multiple concentrations. contacting the (b) providing conditions that allow the formation of new microbe-induced amyloid; and (c) analyzing or quantifying the microbe-induced amyloid formed in (b); method.

[0270] 21. The microbial amyloid or microbial amyloid precursor comprises CsgA, The method of claim 20.

[0271] 22. The method of any one of options 20 and 21, further comprising stirring during (a).

[0272] 23. (a) The contacting is carried out in the presence of an indicator of amyloid formation. A method according to any one of options 20 to 22.

[0273] 24. The indicator is a fluorescent indicator, a spin-label indicator, or a colorimetric indicator. 23. The method according to claim 23.

[0274] 25. The method of option 23 or 24, wherein the indicator is thioflavin T.

[0275] 26. A method for selecting a CsgA- and α-synuclein-containing antibody, wherein the CsgA and α-synuclein are separately labeled. The method according to any one of claims 20 to 25.

[0276] 27. The formation of the above is intrinsic fluorescence, fluorescence of a dye or label, fluorescence resonance energy transfer, fluorescence Light polarization, fluorescence polarization transfer, UV / Vis spectroscopy, magnetic resonance, Raman scattering, electron paramagnetic spin resonance, optical microscopy, electron microscopy, scanning tunneling microscopy, or atomic force microscopy 27. The method according to any one of options 20 to 26, wherein the method is analyzed or measured by

[0277] 28. The method of any of options 20-27, wherein the composition comprises a mixture of compounds. .

[0278] 29. Any of options 20-28, wherein the composition comprises tissue, body fluid, or an extract thereof. The method described below.

[0279] 30. The composition contains feces, urine, blood, cerebrospinal fluid, or saliva, or components thereof. A method according to any one of options 20 to 29.

[0280] 31. The composition comprises one or more bacteria, bacterial extracts, lysates, conditioned culture media, frozen culture media, or the like. Contains dried bacteria, lyophilized lysate, lyophilized culture medium, or any combination thereof. A method according to any of options 28 to 30.

[0281] 35. The method of claim 1, further comprising identifying or selecting a composition that reduces amyloid formation. The method according to any one of claims 20 to 34.

[0282] 36. In (c), the rate of amyloid formation is analyzed or quantified. 6. A method according to any one of 5.

[0283] 37. Contains microbial amyloid or microbial amyloid precursor and α-synuclein A kit comprising a microbial amyloid or microbial A kit in which amyloid precursors and alpha-synuclein are present.

[0284] 38. The microbial amyloid or microbial amyloid precursor comprises CsgA. The kit described in Item 37.

[0285] 39. A method of treating or inhibiting an amyloid disorder in a subject, comprising: (a) detecting a plurality of concentrations of microbial amyloid or microbial amyloid pre-inhibitory agent in the presence of the composition; contacting the precursor with multiple concentrations of alpha-synuclein; (b) analyzing or measuring new amyloid formation after the reaction described in (a); (c) By comparing the analysis or measurement performed under (b) with a control analysis or measurement. wherein the control is the form of amyloid after the reaction described in (a) in the absence of the composition. comparing, including analyzing or measuring the composition; and (d) the formation of amyloid in the presence of the composition is comparable to the formation of amyloid in the absence of the composition. When the formation of amyloid is increased compared to that of amyloid, the compound is suitable for inhibiting or treating the above amyloid disorder. administering to the subject an effective amount of the pharmaceutical composition.

[0286] 40. The microbial amyloid or microbial amyloid precursor comprises CsgA. The method described in Item 39.

[0287] 41. Any of options 39-40, wherein the composition comprises tissue, body fluid, or an extract thereof. The method described below.

[0288] 42. The composition contains feces, urine, blood, cerebrospinal fluid, or saliva, or components thereof. A method according to any one of options 39 to 41.

[0289] 43. Options 39-42, wherein the pharmaceutical composition comprises one or more probiotic bacteria. The method described below.

[0290] 44. The pharmaceutical composition contains Bacteroides, Prevotella Prevotella, Parabacteroides, Faecalibacterium, Eubacterium bacterium, Roseburia, Blautia ), Coprococcus and Bifidobacterium bacterium), or any combination thereof. The method according to options 39 to 43, which includes the above bacteria.

[0291] 45. The pharmaceutical composition is effective against B. fragilis, B. vulgata B. vulgatus and B. thetaiotaomicron omicron), or any combination thereof The method of any one of options 39 to 44, comprising bacteria.

[0292] 46. ​​In (b), the rate of amyloid formation is analyzed or quantified. 6. A method according to any one of 5.

[0293] 47. Identifying or selecting a subject as described above as a subject who would benefit from treatment or inhibition of an amyloid disorder. 47. The method of any one of options 39 to 46, further comprising selecting:

[0294] 48. The above subjects are diagnosed with Parkinson's disease, dementia with Lewy bodies, incident Lewy body disease, Lewy body variant of Alzheimer's disease, multiple system atrophy, pure autonomic failure, or any of these Subjects who are at risk for or exhibit symptoms of any combination of one or more of these conditions 48. The method of any one of options 39 to 47, further comprising identifying or selecting by [Example]

[0295] Example 1 A subject is administered one or more of the compounds (or compositions thereof) described herein periodically. For example, daily oral or rectal administration is recommended. The level of α-synuclein aggregation in tissues can be monitored by stool sampling or biopsy. Prevent bacterial amyloid (curli) formation and / or α-synuclein aggregation Continue treatment to monitor the patient for changes in GI function and motor symptoms. A subject in which administration of one or more compounds results in a reduction in the formation of microbial-induced amyloid in the gastrointestinal tract. Improvement of one or more GI symptoms, one or more motor symptoms, and / or one or more neurological symptoms Goodness is observed.

[0296] Example 2 Obtain or synthesize one or more of the compounds described herein and administer them at the site of action. Enteric-coated or colon-selective formulations are used to release substances in the stomach and bypass most of the small intestine. This allows the incorporation of this molecule into sites of curli production and / or α-synuclein aggregation. This allows for the delivery of the composition in a manner that minimizes absorption of the composition into the systemic circulation.

[0297] Example 3 One or more of the compounds described herein may be obtained or synthesized and administered to the lower small intestine or colon. Incorporate into a formulation for controlled release in the presence of steroids, which allows for lower and / or less frequent Controlled release in the lower small intestine or colon allows for administration of the drug with minimal side effects. This can be achieved by any of a variety of approaches known in the art, including enteric coating. Integral capsules, tablets, soft gels, internal enteric capsules, multi-layer formulations, coated with polymer materials Examples include coated microparticulate forms.

[0298] Example 4 The subject is administered a combination of two or more of the compounds described herein. Combining li inhibitors with α-synuclein aggregation inhibitors simultaneously inhibits aggregation at two critical points. Administration of one or more of the above compounds inhibits the formation of microbial-induced amyloid in the gastrointestinal tract. For subjects who experience a reduction in one or more GI symptoms, one or more motor symptoms, and / or one Improvement in the above neurological symptoms is observed or measured.

[0299] Example 5 WO 2018 / 213204, the entire contents of which are incorporated herein by reference. in curli-driven pathophysiology in mice using techniques described in We first investigated the role of functional amyloid formation in wild-type E. coli biofilm growth. The effect of epigallocatechin gallate (EGCG) on α-Syn amines in vitro was investigated. The effect of EGCG on rhodopsin formation was also investigated. Figure 1A shows the effect of EGCG at the indicated concentrations on rhodopsin formation. The optical density (OD) of wild-type E. coli biofilms grown after 4 days of static incubation was used. 1B is a graph showing crystal violet staining with data evaluated by With or without treatment with EGCG (50 μM), αSyn amyloid was significantly increased alone or was formed in the presence of CsgA (25:1 molar ratio) while thioflavin T fluorescence 1 is a graph showing measured in vitro αSyn aggregation.

[0300] Example 6 WO 2018 / 213204, the entire contents of which are incorporated herein by reference. Additional experiments using the techniques described in

[10] demonstrated that curli were monocolonized with sufficient bacteria. showed that αSyn-induced pathology and increased inflammatory responses in the brain. GF) wild-type (WT) or Thy1-αSyn (ASO) mice were cultured in wild-type, curli either curli-sufficient E. coli (WT) or curli-deficient E. coli (ΔcsgBAC) Figure 2A shows the expression of β-lactamase in whole brain lysates quantified by ELISA. Figure 2B is a graph showing total αSyn in the striatum by dot blot assay. 2C and 2D are graphs showing the quantification of insoluble αSyn fibrils from the striatum. Quantification of TNFα (Figure 2C) and IL-6 (Figure 2D) by ELISA is shown. Figure 2G shows thin brain sections from ASO mice stained for Iba1 (microglia). Staining and Quantifying Morphological Features of Microglia Residing in the Striatum, Generating 3D Cellular Reconstructions The results are shown below. n = 3 (Fig. 2A-2B), n = 6-7 (Fig. 2C, Fig. 2D), n = 4 (Figure 2E-2G) (from 20-40 cells for diameter or 5-10 cells for branching) (Averaged from seven cells). Points represent individuals, and bars represent the mean and standard error. Figure 2A-B. 2D is a one-way ANOVA with Tukey post hoc test, or Figures 2E and 2F are two-tailed t Data analyzed by one-way ANOVA with the *p≦0.05, **p≦0 .01, ***p≦0.001, ****p≦0.0001. Consistent with this effect of rli-sufficient bacteria, the relative abundance of csgA is associated with the development of human Parkinson's disease. It has been shown that csgA is increased in the gastrointestinal tract of patients with PD. Fecal samples (ENA accessions: PRJNA268515, PRJEB4927 and PICRUSt analysis of 16S RNA data available from Based on this analysis, the relative abundance of csgA was determined to be higher in the gastrointestinal tract of PD patients. Furthermore, the expression of wild-type (Fig. 2I) or Thy1-αSyn was significantly higher than that of wild-type (Fig. 2H). (ASO) (Figure 2J) Mice were injected with either human PD or matched controls (ENA accession number 1111111). When colonized with microorganisms derived from the 16s r PICRUSt imputation analysis of RNA sequences revealed a significant increase in the PD transplant microbiota compared to healthy controls In Figures 2H-2J, points represent individuals, and bars represent individuals. *p≦0.05. **p≦0.01. Thus, the presence of bacterial proteins such as csgA in the gastrointestinal tract Presence or elevated levels (compared to healthy controls) correlate with amyloid disorders such as PD. This can be seen.

[0301] Example 7 Further experiments demonstrate that intestinal curli promote progressive synuclein-dependent pathophysiology. Conventionally raised Thy1-αSyn (ASO) animals were given 30 μg of The adult CsgA hexamer (CsgA; N-QYGGNN-C) or the non-amyloidogenic peptide ( N-QYGGNA-C; N-QYGGNA-C) were injected into the intestine. Main steps: beam crossing (Figure 3A), pole lowering (Figure 3B), adhesive removal (Figure 3C) , hindlimb clasping score (Figure 3D), wire hang (Figure 3E), and fecal excretion assessment (70 In the study (Fig. 3F), the locomotor and muscular functions were examined over time at 0, 7, 21, and 70 days after injection. and GI function. Figure 3G is a graph showing a principal component analysis of the compiled motor scores of Figures 3A-3F. Figures 3H to 3I show the results of dot blot assays in the striatum (Figure 3H) and ventral Quantification of insoluble αSyn fibrils in the midbrain (Figure 3I). n = 8 (Figures 3A-G) ), n = 4 (Fig. 3H). Points represent individuals, and bars represent the mean and standard error. As shown above, two-way ANOVA with Sidak post hoc test for between-group comparisons The data in Figure 3H are analyzed using a two-tailed method. Analyzed by Whitney test. *p≦0.05, ** in Figures 3A to 3I. p≦0.01, ***p≦0.001, ****p≦0.0001. Exercise performance measured by PCA Force editing resulted in a symptomatic shift in mice injected with the CsgA peptide compared to controls demonstrating that the overall motor function of these animals is impaired (Figure 3G). Furthermore, the formation of αSyn fibrils in the midbrain of amyloidogenic CsgA-injected animals (Figure 3H) Increases were detected and correlated with central nervous system (CNS) pathology following direct administration of amyloid to the GI tract. Thus, the CsgA peptides capable of forming amyloids Intestinal exposure to αSyn is sufficient to exacerbate long-lasting motor impairment in αSyn-overexpressing mice minutes.

[0302] Thus, as used herein, the intestinal curli are defined as having a crossing time, a descent time, a removal time, and a posterior Increased paw score and decreased drop time and fecal pellets per mouse The increase in crossing time, descent time, and hind limb score, as well as fecal matter per mouse The reduction in pellets was statistically significant at the levels described (Figures 3B-3D and ). 3F). Thus, intestinal curli may mediate symptoms of amyloidosis in vivo. It is thought that this may be induced.

[0303] Example 8 Obtain a library of potential amyloidogenesis inhibitors. Such a library can be used to or can be synthesized de novo, e.g., by combinatorial synthesis. or by solid phase peptide synthesis using methods well known in the art. For example, solid phase peptide synthesis, combinatorial peptide synthesis, and peptide synthesis can be performed. Jensen, et al., herein incorporated by reference for teachings on generating libraries. KJetal.,eds,Peptide Synthesis and Appl. See ications (2nd ed.), 2913. In multi-well assay plates, each well can contain a different ratio of amyloid. Bacterial amyloidogenic bacteria such as E. coli CsgA contain amyloid activator and amyloid precursors. IgE-initiators were added at different concentrations in one dimension to induce the synthesis of host-derived amyloids such as α-synuclein. The idiotype-forming protein is added in different amounts in the other dimension. Add an indicator of amyloid formation, such as ThT, and a fixed amount of each test compound. Each tray is agitated to initiate amyloid formation and thioflavin fluorescence is monitored. Compounds that exhibit a shift in fluorescence generation rate over time are candidates for promoting or inhibiting amyloid formation. is identified as

[0304] Example 9 The suspected amyloid formation inhibitors were analyzed by inducing bacterial amyloidogenesis in the presence of thioflavin T (ThT). Amyloid activators and amyloid precursors were combined. The amyloid initiator, amyloid precursor, and thioflavin T were used as suspected inhibitors of amyloid formation. Combine in the absence of Thioflavin T. Monitor Thioflavin T fluorescence over time. a reduced rate of increase in Thioflavin T fluorescence in samples containing suspected inhibitors compared to and / or a decrease in the maximum level of Thioflavin T fluorescence indicates that the suspected amyloidogenic inhibitor This confirms that it actually functions to inhibit amyloid formation.

[0305] Example 10 A tissue, body fluid, feces, or intestinal content sample is collected from the subject. amyloid initiators, e.g., E. coli CsgA, host-derived amyloidogenic proteins, e.g., , α-synuclein and amyloid formation indicators, such as thioflavin T (ThT), Combine. Monitor Thioflavin T fluorescence. The fluorescence intensity was consistent with an increased rate of amyloid formation in the presence of the sample compared to the rate of amyloid formation. Increased light may help prevent alpha-synucleinopathies such as Parkinson's disease and / or dementia with Lewy bodies. This increased risk is further exacerbated by the α-amyloid-related Defined risks for the onset and / or progression of synucleinopathies or other neurodegenerative disorders The results are correlated with the results of conventional neurological examinations to calculate the score.

[0306] Example 11 In a non-binding black plastic 96-well plate, 50 μM to 100 μM α-Synuclein was dissolved in 0.01 M Tris buffer (pH 7.4) or 0.05 M phosphate buffer. In potassium buffer (pH 7.3) in the presence of 12 μM thioflavin T (prepared in water) Incubate purified Cs in 0.05 M potassium phosphate buffer (pH 7.3). gA monomer was added to each well at a molar ratio of 1:10, 1:25, 1:50, or 1:100. The inhibitor compound is added in an appropriate buffer solution based on solubility, e.g., 0.05 M phosphate buffer. Prepare in potassium buffer (pH 7.3) or DMSO. Add 150 µL of final PBS to the wells containing α-synuclein and CsgA. The concentration of each compound depends on the type of compound being screened. Generally, in initial screening, it is expected to be in the range of 1 μM to 200 μM. For details on the addition of such compounds, see the available small molecule libraries. It depends on the type of substance. The wells served as specificity controls or were tested in combination in the absence of potential inhibitors. Place one sterile glass or Teflon bead with a diameter of approximately 1-2 mm into each well. The plate is incubated at 37°C with continuous orbital shaking (approximately 100-250 rpm) for 1 hour. Incubate in an optical microplate reader. Excitation at 440 ± 10 nm and 490 Fluorescence was measured every 1-2 hours at ±10 nm emission. Measurements were taken over a 24-72 hour period. When α-synuclein amyloid is formed, the emission spectrum changes to a sigmoid curve. According to the results, maximum strength is reached in approximately 24-72 hours under these conditions. Thereafter, the emission intensity may decrease as the amyloid becomes insoluble and non-fluorescent.

[0307] Amyloid formation occurs in three stages (see, for example, FIG. 1B): 1) a lag phase where fluorescence intensity is low over the first approximately 0–24 h; (2) from approximately 2–48 h (3) the logarithmic phase, during which the fluorescence intensity increases logarithmically; (4) the logarithmic phase, during which the fluorescence intensity reaches a maximum and decreases for the remainder of the period; Either the insoluble α-synuclein remains unchanged or it precipitates from the solution. The maximum intensity is generally between 24 and 72 occurs over time.

[0308] Aggregation kinetics, measured by thioflavin fluorescence in the presence of compounds, was The kinetics can be normalized to that observed using CsgA and CsgA alone. Potential inhibitors may be used to extend the lag phase, decrease the rate of change in the log phase, It may act to reduce the maximum attainable intensity, or any combination of these.

[0309] Once initial candidates are identified, the dose response over a wide range of concentrations can be determined, and The specificity for CsgA:synuclein aggregates can be determined by and α-synuclein specificity can be determined individually. In some variations of the method, CsgA:synuclein aggregates are monitored up to the log phase. At this point, a potential inhibitory compound can be introduced. Inhibitors that can act during progression of the disease can be identified (see, e.g., Figure 1B). (I want to be).

[0310] Example 12 The assay was performed as described in Example 11, with a full concentration curve for each compound. A line is generated, which allows the EC50 of each compound to be accurately determined. Identify limitations of specific compounds (e.g., compounds that do not result in complete inhibition of aggregation) (to clarify).

[0311] Example 13 The assay was performed as described in Example 11 and was performed in a high-throughput manner. For example, instead of a full concentration curve for each compound, you might want to use 3 A point concentration curve is used to identify compounds with dose-response effects that have nonspecific and concentration-independent effects. Furthermore, for high-throughput screening, the assay The compounds are formatted into 96-well, 384-well, or 1536-well plates. Test at a single concentration (e.g., 100 μM) and a single time point (e.g., 24–72 hours). This allows the observer to differentiate potentially ineffective compounds from compounds that are ineffective or ineffective at the appropriate concentrations. It is possible to distinguish between suitable candidates.

[0312] Example 14 The assay was performed as described in Example 11, and the total time course for each compound was A time course curve is generated that indicates whether the compound inhibits over time in a linear or sigmoidal manner. It indicates whether inhibition is achieved and / or whether complete inhibition can be achieved with a given compound.

[0313] Example 15 The assay was performed as described in Example 11 and assayed for α-synuclein (or other Both the host amyloid protein and Curli (or other bacterial amyloids) are To assess mechanistic processes and compound activity in the more dynamic environment present in assays The observer then examines one or the other of the proteins in the presence of the agglutinated template. Evaluate the ability of compounds to inhibit protein aggregation, e.g., the bacterial amyloid component Csg A is known to promote and / or hasten α-synuclein aggregation. In this study, candidate compounds with α-synuclein aggregation inhibitory activity were identified, which showed that bacterial amyloid Thus, the formation of monomeric α-synuclein (or The assay was formatted to include both bacterial amyloid (other host amyloid) and aggregated bacterial amyloid. By using a 3D mouse model, drug candidates can be evaluated in a more physiologically relevant in vitro environment. This makes it possible.

[0314] Example 16 The assay was performed as described in Example 11, and the method was performed using Curli (bacteria). The method further includes combining an inhibitor of α-synuclein aggregation with an inhibitor of α-synuclein aggregation. This combination has the added advantage of simultaneously inhibiting aggregation at two critical points. This assay detects the monomeric forms of both α-synuclein and Curli (CsgA). Using this, measurements similar to those shown in FIG. 1B are obtained.

[0315] Example 17 The assay was performed as described in Example 11, and the method was performed using Curli (bacteria). Further, a combination of amyloid disaggregation promoters with α-synuclein disaggregation promoters is also contemplated. This combination has the added advantage of simultaneously deagglomerating at two critical points. This assay utilizes fully aggregated forms of both α-synuclein and Curli. Using this method, measurements similar to those shown in Figure 1B are obtained.

[0316] Example 18 The assay was performed as described in Example 11, and the method was performed using curli (Cs Combining an inhibitor of α-synuclein aggregation (gA, bacterial amyloid) with an agent promoting α-synuclein disaggregation This combination inhibits amyloid nucleation or development while also: This assay has the added advantage of providing disaggregation of an already initiated process. , fully aggregated form of α-synuclein and monomeric form of Curli (CsgA). Utilizing this, measurements similar to those shown in Figures 2A-3I are obtained.

[0317] Example 19 The assay was performed as described in Example 11, and the method was performed using Curli (bacteria). Furthermore, the combination of amyloid disaggregation promoters with α-synuclein aggregation inhibitors is This combination destroys pathogenic bacterial amyloid while simultaneously inhibiting alpha-synuclein. This assay has the added benefit of inhibiting α-synuclein monoclonal antibody aggregation. The mer and fully aggregated forms of Curli were utilized, as shown in Figures 2A to 3I. Similar measurements are obtained.

[0318] Example 20 Similar to any of Examples 16-24, except that a non-aggregatable CsgA mutant was included. Assays are also performed to assess the presence of structured (aggregated) CsgA in these processes. Dependent compounds show reduced efficacy in this version of the assay.

[0319] Example 21 Human α-synuclein was synthesized in E. coli BL21(DE3) by transfection with untagged human α-synuclein. The full-length gene of klein was expressed from a plasmid derived from pT7 or pET11a. Cells were induced at OD600 0.6 with 0.8 mM IPTG for 4 hours and then centrifuged. and lysed in a volume of lysis buffer (10 mM Trisodium Thr) equivalent to 1 / 10 of the culture volume. Suspend the cells in PBS (pH 8.0, 1 mM EDTA, and 1 mM PMSF) and boil for 20 min. Cell debris was pelleted by centrifugation and the clarified lysate was The lysate was treated with 136 μL / mL of 10% streptomycin sulfate, followed by 228 μL The lysate was centrifuged, the supernatant transferred to a new tube, and an equal volume of saturated HCl was added. Proteins were precipitated by the addition of ammonium sulfate (100%) and incubated on a rocking shaker. The mixture was incubated at 4°C for 1 hour. The protein was pelleted by centrifugation and Wash with 100 mM ammonium acetate in chilled ethanol and pellet by centrifugation. The cells were then lysed in 50 mM potassium phosphate buffer, washed twice with cold ethanol, dried overnight, and resuspended in 50 mM potassium phosphate buffer. (pH 7.3) or 10 mM Tris (pH 7.4) and resuspended in 10 mM Tris (pH 7.4). Immediately before use, purified α-synuclein was passed through a 0.2 μm nylon filter. The concentration of α-synuclein was determined by absorbance at 280 nm or BCA assay. was determined using

[0320] Example 22 The full-length recombinant CsgA monomer was synthesized by Zhou et al. (2012). al of Biological Chemistry 287(42) Briefly, csgA can be prepared by ligating it with a C-terminal 6xHis tag. After growth in rich medium, CsgA production was confirmed. The live cells were grown to an OD of approximately 0.9 with 0.5 mM IPTG. 600 Induced at 37°C for 1 hour. The bacteria are incubated overnight or for approximately 1-2 hours at room temperature on a rocking platform in 50 mM phosphate buffer. Dissolved in 8 M guanidine hydrochloride in potassium buffer (pH 7.3). 10,000× After centrifugation at 400 × g for 20 min, the supernatant was sonicated six times for 10 s on ice to remove the nickel-nitride. Incubation with ribotriacetic acid resin (Sigma) for 1 hour at room temperature followed by disposable The column was packed in 50 mM potassium phosphate. 50 mM phosphate buffer (pH 7.3) containing 12.5 mM imidazole The protein was washed with potassium buffer (pH 7.3). The monomeric CsgA was eluted with 50 mM potassium phosphate buffer (pH 7.3) containing 100 mM HCl. Combine the fractions containing the target protein and filter through a 30 kDa centrifugal filter to obtain The purified Csg was then placed in a Thermo Scientific HPLC reactor to remove dimers and other oligomers. A was passed through a pre-chilled desalting column (Zeba) to remove imidazole.

[0321] Alternatively, a synthetic hexapeptide of CsgA consisting of the following sequence: Nterm-QYGG NN-Cterm is commercially available from Bio-synthesis, Inc.

[0322] Example 23 Another alternative is Collinson et al. (1991). Journal o f Bacteriology.173(15) as described in the biophil Preformed CsgA antigens were isolated by preparing purified Curli extracts from the Wild-type E. coli was incubated at room temperature for 3 to 7 days in a Congo red dye bath. The cultures were grown on YESCA medium with or without added ATP. Scrape the cells into 0 mL of 10 mM Tris (pH 8). Cells can be sonicated or freeze-thawed. The cell lysate was dissolved in 0.1 mg of RNase A and 0.1 mg of DNase I. and MgCl2 added up to 1 mM, and incubated at 37°C for 20-30 minutes. Add lysozyme to 1 mg / mL and incubate at 37°C for an additional 20-40 minutes. Add SDS to 1% and incubate at 37°C for 20-40 minutes. The supernatant is collected by centrifugation at 12,000 × g for 15 minutes. The sample is collected in 1-10 mL volumes. Resuspend in Tris buffer, boil at 90°C for 15 min, and repeat the above process (R The sample was then washed with Tris buffer (digestion by Nase, DNase, lysozyme and SDS treatment). Wash twice with PBS, resuspend in Laemli buffer, boil, and run on an SDS-PAGE gel (4–2 0%). Electrophorese the samples at 20 mA for 5 hours. The insoluble material was collected, washed three times with water, washed twice with 95% ethanol, and dried. Resuspend the sample in 0.2 M glycine (pH 1.5) and boil for 10-15 minutes. The soluble material is recovered by centrifugation at 16 k×g for 10 minutes. The insoluble material is washed with water. The samples were washed five times and resuspended in PBS. Finally, the samples were sonicated for 1 hour using an electrode or a water bath. After digestion, protein content is determined by BCA or absorbance at 280 nm.

[0323] Such changes may affect the kinetics of aggregation, the concentration of compound required to inhibit aggregation, and the CsgA-mediated signal transduction. The ratio required to show nuclein aggregation, or any combination of the above, can be varied.

[0324] Example 24 In other iterations of the protein purification used in the assays of Examples 22 and 23, C sgA can be produced without the histidine tag or with another tag to regulate its export from the cell. The α-synuclein may contain a sequence that promotes its release. To facilitate purification by affinity for ATP, a histidine tag or other tag may be included. CsgA and α-synuclein can be engineered using alternatives well known to those skilled in the art. Alternative methods, such as ammonium sulfate precipitation using different concentrations of ammonium sulfate in a single step, or It can be purified using ammonium sulfate precipitation in multiple steps with increasing ammonium sulfate concentrations. Alternatively, the ammonium sulfate precipitation may be omitted. based on affinity, cation exchange, anion exchange, hydrophobic interaction, multiple methods or mixed methods Another resin or reagent was used to separate CsgA or α-synuclein from other proteins. Protein separation can be performed using methods well known to those skilled in the art. , batch purification, prepacked column, gravity flow, low pressure, high pressure, and high pressure liquid chromatography These methods can be used individually or in combination. CsgA or α-synuclein was purified by size exclusion chromatography or high-pressure liquid chromatography. Other types of tags can be identified based on size using methods well known to those skilled in the art, such as micrographs and micrographs. CsgA and α-synuclein can be isolated from the protein. For all or part of the process, different concentrations of guanidinium hydrochloride or equivalents may be used under denaturing conditions. It can be purified using other denaturing agents, such as urea, at various concentrations known to those skilled in the art. Alternatively, CsgA and α-synuclein can be purified under native conditions well known to those skilled in the art. CsgA is generally rapidly purified by processes using native conditions or non-denaturing buffers. His-tagged CsgA is purified to avoid aggregation during the purification process. use alternative methods known to those skilled in the art, such as the addition of chelating agents such as ethylenediaminetetraacetic acid. and can be eluted from immobilized metal affinity materials such as Ni-NTA. Immobilized metal affinity chromatography based on the affinity of histidine residues of proteins for metals In addition to or instead of chromatography, as known to those skilled in the art, e.g. histidine or another amino acid in a protein (e.g., cysteine ​​or tryptophan) Materials with interacting immobilized copper, zinc, cobalt, or nickel Another immobilized metal affinity chromatography or batch purification method using the material As known to those skilled in the art, other buffers may be used for Ni-NTA agarose. It can be used with a base such as tris(hydroxymethyl)aminomethane (" Tris”), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (“H EPES), and 3-(N-morpholino)propanesulfonic acid (MOPS). Optionally, sodium chloride, potassium chloride or other salts and Ni-NTA agar Various surfactants of various compositions and concentrations compatible with column chromatography or batch purification and those containing a reducing agent can be used.

[0325] Other concentrations of dimethyl sulfoxide may be used in this assay, but significantly higher Concentrations of dimethyl sulfoxide as low as or less than 1% inhibited the activity of CsgA and α-synuclein. Other concentrations of Thioflavin T may be used in the assay, which may affect aggregation kinetics. , which may affect the fluorescent signal and sensitivity of the assay. α-synuclein may be used, and such modifications may be used to enhance the activity of α-synuclein in the assay. Different concentrations of the compounds were tested in the assay. The dose response may be assessed. This may affect the aggregation kinetics of α-synuclein and CsgA depending on their concentrations. Additional reagents may be added to the assay. Shaking may be included in the assay at different intervals. This may affect the aggregation kinetics of CsgA and α-synuclein. Las beads may be omitted from assays containing α-synuclein or CsgA. Alternatively, beads of other sizes or compositions may be used, These changes may affect the aggregation kinetics of CsgA and α-synuclein. s, other buffers such as HEPES or MOPS, and other buffer concentrations may be used in the assay. These may affect the aggregation kinetics of CsgA and α-synuclein. A fluorescent probe with a sufficiently narrow bandwidth, such as 10 nm, that excites at 38 nM and emits at 495 nm. Any plate reader capable of optical reading can be used. Alternative microplates, such as black microplates, may be used for the assay. The container may be sealed with another coating that does not absorb ThT fluorescence, or the coating may be Fluorescence may be measured at a single endpoint or over various time intervals. Readings may be taken at multiple points, and the time intervals at which fluorescence is measured may be constant; The lag phase of Thioflavin T fluorescence, the fluorescence signal, The shape of the curve generated by the kinetic readout is used to determine the slope of the curve. Other metrics, such as examination of thioflavin T signaling over time, can be used to assess α-synuclein expression. The effect of the compounds on nuclein and CsgA expression can be examined.

[0326] Example 25 To evaluate the effect of compounds on α-synuclein aggregation, purified α-synuclein was A cell-free assay was performed using thioflavin and thioflavin T. In this assay, E. coli In BL21(DE3), the full-length untagged human α-synuclein gene was inserted. Human α-synuclein was expressed from plasmids derived from pT7 or pET11a. The cells were incubated at 0°C for 1 hour with 0.8 mM IPTG. 600 Induce with 0.6 for 4 hours and then centrifuge The cells were collected and lysed in a volume of lysis buffer (10 mM Tris (p Suspend the cells in 100% ethanol (H8.0), 1 mM EDTA, and 1 mM PMSF) and boil for 20 minutes. Cell debris was pelleted by centrifugation and the clarified lysate was collected by 1 Treatment with 36 μL / mL of 10% streptomycin sulfate followed by 228 μL / mL The lysate was centrifuged, the supernatant was transferred to a new tube, and an equal volume of saturated (100 Proteins were precipitated by the addition of 1% ammonium sulfate and incubated for 1 h at 4 °C on a rocking shaker. The protein was pelleted by centrifugation and diluted with an equal volume of cold ethanol. Wash with 100 mM ammonium acetate in ethanol, pellet by centrifugation, and cool. Wash twice with ethanol, dry overnight, and then resuspend in 50 mM potassium phosphate buffer (pH 7.3 ) or 10 mM Tris (pH 7.4) and loaded onto a 50 kDa cutoff column. Immediately before use, the purified α-synuclein was filtered through a 0.2 μm nylon filter. The assay consisted of one 1-2 mm glass bead, 20-40 μM thiazol-3, and 10 μM thiazol-3 in each well. Ofravin T, 1% DMSO, 50 μM α-synuclein, and 20-100 μM phosphate buffer. The experiment was carried out in a 96-well black microplate containing the mixture. The plate was sealed with sealing tape (Th The tube was sealed with a thermocouple (Fisher 232701) and then placed on a Tecan Nano F200 plate. Incubate at 37°C in a chromatograph and filter using a 439 nm filter with a bandwidth of 8 nm. Excitation at 438 nm and emission at 495 nm using a 490 nm filter with a 10 nm bandwidth During the assay, the plates were shaken continuously or shaken for 999 seconds every 18 minutes. Readings were taken every hour for up to 73 hours. The effect of compounds on nuclein aggregation was assessed by deconvoluting at 36, 47, 48 or 72 hours. The time before or approximately the time when ThT fluorescence plateaued in the data or vehicle control wells. The vehicle control was determined using data from the final assay time point closest to the time of administration. The assay contains all components of the assay except the compound and measures α-synuclein in the absence of inhibitors or enhancers. The fluorescence value of each compound with α-synuclein was first calculated by adjusted by subtracting the mean fluorescence of that compound in the absence of α-synuclein These adjusted values ​​were divided by the mean fluorescence at the same time points in vehicle control wells. The mean normalized fluorescence due to the compound was then normalized to the mean fluorescence observed in the untreated controls. The inhibition of α-synuclein aggregation by compounds was expressed as a percentage of the mean vehicle vs. The percentage of mean fluorescence due to the compound relative to illumination is calculated by subtracting it from 100%. A higher positive inhibition rate reflects greater inhibition of α-synuclein aggregation. On the other hand, the negative inhibition rate reflects the enhancement of α-synuclein aggregation.

[0327] In some embodiments, other variations in protein purification are used in the assay. Alternatively, α-synuclein can be purified by alternative methods well known to those skilled in the art, e.g., in a single step. It can be purified using ammonium sulfate precipitation with different concentrations of ammonium sulfate, or In some embodiments, the acid ammonium salt may be omitted, as is well known to those skilled in the art. - Separating synuclein from other proteins with a separate resin or device (e.g., protein (based on protein affinity, cation exchange, anion exchange, or hydrophobic interactions) Multiple or mixed modes may be used. In some embodiments, the purification may be carried out by batch purification, packed columns, gravity flow, or other methods using methods well known to those skilled in the art. Protein separation can be performed using low-pressure, high-pressure and high-pressure liquid chromatography. These methods can be used individually or in combination. The method is well known to those skilled in the art, such as size exclusion chromatography or high pressure liquid chromatography. It can be separated from other proteins by size using known methods. Alpha-synuclein is purified under various denaturing conditions during all or part of the assay purification process. guanidinium hydrochloride at various concentrations or other denaturing agents known to those skilled in the art, such as urea Alternatively, alpha-synuclein can be purified using methods such as, but not limited to, human alpha-synuclein. Purification can be performed using affinity tags such as stigma tag, BAP tag, or GST tag. As known to those skilled in the art, examples of suitable hydroxybenzoates include, but are not limited to, tris(hydroxybenzoates). (Dimethyl)aminomethane ("Tris"), 4-(2-hydroxyethyl)-1-piperane diethanesulfonic acid ("HEPES"), 3-(N-morpholino)propanesulfone Use of alternative buffers such as phosphate-buffered saline ("MOPS") or phosphate-buffered saline Optionally, sodium chloride, potassium chloride or other salts and protein refinement may be added. Various surfactants and reducing agents of varying compositions and concentrations compatible with the manufacturing process can be used. can.

[0328] Although other concentrations of dimethyl sulfoxide may be used in the assays described herein, Concentrations of dimethyl sulfoxide significantly higher or lower than 1% inhibited α-synuclein production. Different concentrations of Thioflavin T were used in the assay. Different concentrations of α- Such modifications may be used to identify alpha-synuclein in assays. Different concentrations of the compound may be tested in the assay, and the dose Response may be assessed. Sodium dodecyl sulfate, sodium chloride, dithiothreitol or containing detergents such as bovine serum albumin, depending on the concentration of α-synuclein. Additional reagents that may affect the kinetics of aggregation may be added to the assay. The shaking may be included in the assay at different intervals and may affect the aggregation kinetics of α-synuclein. Teflon beads of various sizes may be included in the assay, or other compositions may be used. beads may be used, and these changes affect the aggregation kinetics of α-synuclein. Other buffers such as Tris, phosphate buffered saline, HEPES, and MOPS, and Alternative buffer concentrations may be used in the assay, which may affect the aggregation kinetics of α-synuclein. It excites at 438 nM and emits at 495 nm, and a sufficient amount of Any plate reader capable of fluorescent readings with a narrow bandwidth can be used. Examples include Spectra Max M5, SpectraMax M2 or Sp ectraMax® i3X (Molecular Devices, California) (San Jose, Calif.) Black microfiber with a clear bottom. Black plates, solid white microplates, or white microplates with a clear bottom Other microplates may be used for the assay, such as 384-well or 1536-well plates. Different plate densities can be used, such as well formats. , including, but not limited to, non-bonding coatings or high-bonding coatings. They may be treated or they may be untreated. The sample may be sealed with a separate coating that does not absorb the radiation, or the coating may be removed before reading. Fluorescence can be measured at a single endpoint or at multiple points over various time intervals. The time intervals at which readings may be taken and fluorescence measured may be constant or may vary over the time of the assay. The results are shown in Tables 3 and 4. The higher the positive inhibition rate, the higher the α The negative inhibition rate reflects a greater inhibition of α-synuclein aggregation, whereas the negative inhibition rate reflects a greater inhibition of α-synuclein aggregation. Reflects enhanced klein aggregation.

[0329] Example 26 In vitro thioflavin T assay to investigate the effect of compounds on CsgA aggregation To evaluate the effect of compounds on the aggregation of E. coli CsgA, purified CsgA and thiol were used. A cell-free assay was performed using flavin T. In this assay, a histidine-tagged CsgA was expressed in E. coli NEB30 harboring the pET11d vector containing a C-terminal 6xHis tag. The 16slyD::kan cells were overexpressed in 0.5 mM slyD::kan after growth in rich medium. followed by the addition of IPTG (isopropyl beta-D-1-thiogalactopyranoside) Incubate at 37°C for 1-4 hours with shaking at 200-250 rpm. , an O.D. of approximately 1.0 600 CsgA production is induced by 50 mM potassium phosphate buffer. Resuspend in 8 M guanidine hydrochloride in buffer (pH 7.3) and rinse on ice for 20 seconds at intervals of 3 times. Lysates were lysed by sonication. The resulting lysates were incubated for 1 hour at room temperature on a platform rocker. After centrifugation at 10,000 × g for 20 minutes, the supernatant was placed on ice. The mixture was sonicated three times at 20-second intervals, and then coated with nickel-nitrilotriacetic acid resin (Sigma). ) in a 50 mL conical tube on a platform rocker for 1 hour at room temperature. The resin was washed and then diluted with 50 mM potassium phosphate buffer (pH 7.3). The resin was further washed with 45 mL of 8 M guanidine and pelleted five times with 12.5 mM guanidine. 8M guanidine hydrochloride in 50mM potassium phosphate buffer pH 7.3 containing midazole The purified CsgA protein was pelleted twice with 250 mM imidazole. 6-8 M guanidine hydrochloride in 50 mM potassium phosphate buffer (pH 7.3) The eluted protein was aliquoted at 150 μL per aliquot. The resulting mixture was then rapidly frozen in liquid nitrogen and stored at -80°C. The purified CsgA protein was thawed and passed through two pre-chilled desalting columns (Zeba). The midazole and guanidine were removed. The buffer-exchanged proteins were filtered through a 30 kDa centrifuge filter. The mixture was placed in a Turbo Unit (Amicon) and spun at 7500×g for 30 minutes to separate the dimers and Other oligomers were removed. Protein concentration was determined using NaCl as is well known to those skilled in the art. CsgA was assessed using noDrop technology (Thermo Scientific). Aggregation assays were performed using 20 μM to 40 μM thioflavin T in 50 mM potassium phosphate buffer. pH 7.3, 1% DMSO, 2 μM to 20 μM CsgA, and 3.125 μM 96-well black non-binding surface or white untreated microplates containing ~200 μM compound Vehicle control wells contained all components of the assay except the test compound, The plates were plated using a Tecan plate reader. A 439 nm filter with an 8 nm bandwidth was used in a Nano F200 plate reader. Excitation was performed at 438 nm using a 490 nm filter with a 10 nm bandwidth. Spec excitation at 438 nM and emission at 495 nm, or SpectraMaxM5, SpectraMaxM2 or SpectraMax (registered trademark) ) i3X (Molecular Devices, San Jose, CA) spectrophotometer During the assay, the plate was first shaken for 5 seconds, followed by Fluorescence readings were taken every 15 minutes for up to 1005 minutes after shaking for 3 seconds. The effect of compounds on CsgA aggregation in the assay was evaluated using data from 0 to 17 hours. The fluorescence value of each compound containing CsgA was first measured at 0 or 15 minutes. The mean fluorescence of the compound in the absence of CsgA or in the presence of CsgA and the compound. The data were adjusted by subtracting the mean fluorescence of the combined data. did.

[0330] 1. Divide the adjusted ThT fluorescence value by the mean fluorescence of the vehicle control wells containing CsgA. Normalized fluorescence from 6-6.5 hours or 8-9 hours with this compound. The median light was expressed as a percentage of the median fluorescence observed in the vehicle control over the same period. The percentage of inhibition of CsgA aggregation by a compound was expressed as a ratio of the compound to the vehicle control. The percentage of mean fluorescence due to α-glucan was calculated by subtracting it from 100%. The percent inhibition reflects a greater inhibition of CsgA aggregation, while the percent negative inhibition reflects a greater inhibition of Cs Reflects enhanced gA aggregation.

[0331] 2. GraphPad Prism (GraphPad Software, Sandy) Adjustments for each compound concentration and vehicle control were made using a 100% ethanol-based ... The measured ThT fluorescence values ​​were plotted as a progress curve over time. At 55, 510, 750, and 1005 minutes, the vehicle control and each test concentration were The area under the curve (AUC) for the compound was determined. Percentage of control AUC was calculated for each test at a given time point. The mean AUC for each compound at the test concentration was calculated by dividing the mean AUC for the vehicle control at the same time point. The percentage of lower AUC of the control represents the Cs reduction by the compound at a given concentration and time point. While the higher percent of control AUC reflects greater inhibition of gA aggregation, This reflects less inhibition of CsgA aggregation by the compound at a given concentration and time point.

[0332] 3. Adjusted ThT fluorescence values ​​for each compound concentration and vehicle control are plotted using GraphP The progress curves were plotted using ad Prism. The progress curves were calculated by the Hill slope (Y = B max *X^h / (T 1 / 2 GraphP for specific binding using ^h+X^h) The adPrism equation was used for fitting. The binding maximum, i.e., B max ThT The maximum specific binding is expressed as the fluorescence. The time to half-maximum binding (T 1 / 2 ) is half maximum B max is the time required to achieve fluorescence, and the higher the T 1 / 2 indicates that the compound is Delaying CsgA aggregation in the assay or extending the lag phase of CsgA aggregation in the assay When ThT molecules bind to a single site on CsgA without cooperativity, the Hill slope (h) is equal to 1.0. If the Hill slope is greater than 1.0, CsgA exhibits positive cooperativity. It has multiple binding sites for ThT with different affinities for CsgA. If there are multiple binding sites or if there is negative cooperativity, the Hill slope is less than 0. For each test concentration of the substance, a vehicle control B max % is the average B of the compound max A vehicle Control mean B max Divide by 100% and multiply by 100%. Vehicle control B max The compound inhibited CsgA aggregation more significantly in the assay. The values ​​obtained for each parameter are plotted using GraphPad Pris The equation [inhibitor] vs. response was replotted using a variable slope (four parameters) and the data IC 50 reported as.

[0333] In another embodiment, the protein purification used in the assay for CsgA is performed using histidine dinucleotides. They can be produced without a tag or with another tag, and they can be used to detect their export from the cell. CsgA protein induction can be observed in different cells at different times and temperatures. CsgA can be induced in cultures using various cell densities and concentrations of IPTG. Alternatively, alternative methods well known to those skilled in the art, such as adding different concentrations of ammonium sulfate in a single step, Ammonium sulfate precipitation using ammonium sulfate or multiple steps with increasing ammonium sulfate concentrations In some embodiments, the ammonium sulfate precipitation may be omitted. In some embodiments, the binding site may be selected from a variety of binding sites, including protein affinity, cation affinity, etc., as is well known to those skilled in the art. CsgA based on exchange, anion exchange, hydrophobic interaction, multiple methods or mixed methods Other resins or materials may be used to separate the protein from other proteins. Separations can be performed using methods well known to those skilled in the art, such as batch purification, packed columns, gravity flow, etc. These methods can be performed using low-pressure, high-pressure, and high-pressure liquid chromatography. The methods can be used individually or in combination. CsgA can be isolated by size exclusion chromatography. using methods well known to those skilled in the art, such as chromatography or high pressure liquid chromatography. CsgA can be separated from other proteins by size. For all or part of the manufacturing process, different concentrations of guanidinium hydrochloride or It can be purified using various concentrations of other denaturing agents, such as urea, well known to those skilled in the art. In some embodiments, CsgA can be expressed under natural conditions well known to those of skill in the art. CsgA is purified under conditions and isolated by a process using native conditions or non-denaturing buffers. Generally, purification is rapid to avoid aggregation during the purification process. His-tagged CsgA is purified at pH Alternative methods known to those skilled in the art include reducing the amount of ethylenediaminetetraacetic acid or adding a chelating agent such as ethylenediaminetetraacetic acid. The method can be used to elute from immobilized metal affinity materials such as Ni-NTA. Immobilized metal affinity based on the affinity of histidine residues in proteins for immobilized nickel In addition to, or instead of, chromatography, in some embodiments, As known to those skilled in the art, for example, histidine or other amino acids of proteins (e.g., Immobilized copper, zinc, and cobalt interact with cysteine ​​or tryptophan or immobilized metal affinity chromatography using immobilized nickel as a material. A feed or batch purification method can be used. In some embodiments, a separate buffer is used. The solution can be used with Ni-NTA agarose, e.g., Tris(hydroxymethyl) Tris)aminomethane ("Tris"), 4-(2-hydroxyethyl)-1-piperazine Ethanesulfonic acid ("HEPES") and 3-(N-morpholino)propanesulfone Acid ("MOPS"), optionally with sodium chloride, potassium chloride or other salts; and a composition compatible with Ni-NTA agarose chromatography or batch purification; Various concentrations of detergents and reducing agents can be used in this assay. In this case, other concentrations of dimethyl sulfoxide may be used, but not significantly higher or 1 percent Concentrations of dimethyl sulfoxide below 100 mg / ml may affect the aggregation kinetics of CsgA. Different concentrations of Thioflavin T may be used in the assay, which may increase the fluorescence signal of the assay. Other concentrations of CsgA may be used, and such variations may affect the sensitivity and robustness of the assay. Further addition of compounds may affect the aggregation kinetics of CsgA in the assay. May be tested in assays and dose response assessed. Sodium dodecyl sulfate, Na of CsgA, including detergents such as Cl, dithiothreitol, or bovine serum albumin. Adding additional reagents to the assay that may affect their aggregation kinetics depending on their concentration Agitation by shaking may be included in the assay at different intervals to enhance aggregation of CsgA. The 2 mm glass beads or Teflon beads containing CsgA may affect the aggregation kinetics. Beads of other sizes or compositions may be included in the assay, and these Changes in the concentration of Tris, HEPES, and MOPS can affect the aggregation kinetics of CsgA. Any other buffer and other buffer concentration may be used in the assay, including CsgA It excites at 438 nM and emits at 495 nm. Use any plate reader capable of fluorescence reading with a sufficiently narrow bandwidth, such as a Black microplates with clear bottoms, solid white microplates Alternatively, use a different microplate for the assay, such as a white microplate with a clear bottom. Different plate densities, such as 384-well or 1536-well formats, may be used. Plates may also be used with, but not limited to, non-bonded coatings. They may also be treated with a coating such as a high-bond coating. The plate may be sealed with another coating that does not absorb ThT fluorescence. Alternatively, the coating may be removed prior to reading. Fluorescence may be measured at a single endpoint, or The time interval at which fluorescence is measured may be read at multiple points over various time intervals. may be constant or may vary over the time course of the assay. The results are shown in Table 3 and shown in Table 4. Lower Control B max Percent or lower control AUC percent , reflecting greater inhibition of CsgA aggregation, while the higher control B max percent A lower or higher percent control AUC reflects less inhibition of CsgA aggregation. do.

[0334] Example 26A In vitro testing to investigate the effects of compounds on CsgA-seeded α-synuclein aggregation Effect of compounds on aggregation of Escherichia coli CsgA-seeded α-synuclein in the Ofravin T assay Using purified alpha-synuclein, purified CsgA, and thioflavin T to assess A cell-free assay was performed in which histidine-tagged CsgA was introduced into the C-terminus of the A sequence of six histidine residues is added, and it contains the Sec signal sequence (amino acids 1 to 22). E. coli NEB3016slyD carrying the pET11d vector containing missing csgA CsgA was overexpressed in ::kan cells. To induce overexpression of CsgA, Optical density (OD 600 ) was added to the culture at 0.8 to 1 mM IPTG for induction. The cells were incubated at 37°C for 1 hour and then harvested by centrifugation. Below, 8 M guanidine hydrochloride in 50 mM potassium phosphate buffer (pH 7.3) was used. Dissolve CsgA for 1-2 hours at room temperature on a rocking platform or overnight at 4 °C. Batch purification using Nickel-NTA agarose and low pressure and disposable polypropylene columns Immobilized metal affinity chromatography in combination with gravity flow through the column. Therefore, under low pressure applied manually by applying a syringe plunger to the column, Washing with 50 mM potassium phosphate buffer (pH 7.3) followed by 50 mM Washing with 12.5 mM imidazole in potassium phosphate buffer (pH 7.3) and 125 mM imidazole in 50 mM potassium phosphate buffer (pH 7.3) The product was purified via immobilized metal affinity chromatography, including elution with imidazoline. The buffer containing ethanol was freshly prepared before protein purification. The solution was passed through a 1000 kJ / mL molecular weight cutoff filter and then through a desalting column. All steps in the purification of CsgA in the absence of salt were carried out in rapid succession with as little delay as possible. CsgA was measured by BCA assay or by 280°C using a Nanodrop spectrophotometer. Human α-synuclein was quantified using absorbance at 100 nm. Among them, pT7 or pET1, into which the full-length untagged human α-synuclein gene was inserted, Expression was from a plasmid derived from 1a. Cells were grown to OD with 0.8 mM IPTG. 60 The cells were induced at 0.6 for 4 hours. The cells were lysed by boiling for 20 minutes and the cell debris was removed. The cells were pelleted by centrifugation and the clarified lysate was added to 136 μL / mL of 10% sulfuric acid suspension. Treat with streptomycin, add 228 μL / mL of glacial acetic acid, and then centrifuge the lysate. The supernatant was transferred to a new tube and purified by adding an equal volume of saturated (100%) ammonium sulfate. The protein was precipitated. The ammonium sulfate pellet was dissolved in an equal volume of 100 mM ethanol. Wash with ammonium acetate, pellet by centrifugation, and wash twice with ethanol. Dry overnight and resuspend in 10 mM Tris (pH 7.4) with a 50 kDa cutoff. Immediately before use, the purified α-synuclein was filtered through a 0.2 μm nylon filter. The assay consisted of one 1-2 mm glass bead per well, 20-4 0 μM Thioflavin T, 1% DMSO, 2 μM CsgA, 50-60 μM α-CsgA Nuclein, 100 mM sodium chloride, 9.3 mM potassium phosphate (pH 7.3) The experiments were carried out in a 96-well black microplate containing 50 μM of compound. The plate was sealed with sealing tape (ThermoFisher 232701). The plates were then analyzed in a Tecan Nano F200 plate reader at 37°C with continuous shaking. Excitation was performed at 438 nm using a 439 nm filter with a bandwidth of 8 nm, and a 4 Emitting at 495 nm with a 90 nm filter, or using a SpectraMax M5 or Excitation was at 438 nM in SpectraMax® i3X, and emission was at 495 nm. The assay was incubated at 4°C for 1 hour and readings were taken every hour for up to 73 hours. The effect of compounds on CsgA-seeded α-synuclein aggregation in mice was assessed at 16 to 52 hours. Data from within 100 time points were used to investigate the effect of each compound with CsgA and α-synuclein. The fluorescence value of a compound was first calculated by measuring the average fluorescence intensity of that compound in the absence of CsgA and α-synuclein. These adjusted values ​​were used for all assays except for compound. The positive value is calculated by dividing by the mean fluorescence at the same time point in the component-containing vehicle control wells. Normalized and mean or median normalized fluorescence due to compound observed in vehicle control at the same time point The fluorescence was expressed as a percentage of the mean or median fluorescence obtained. Inhibition of synuclein aggregation was calculated by subtracting the mean percent or median fluorescence at 100% compound. A higher positive inhibition rate was obtained for CsgA-seeded α-synuclein aggregation. While the negative inhibition rate reflects a greater inhibition of CsgA-seeded α-synuclein Reflects increased aggregation.

[0335] [Table 3] TIFF2026016368000062.tif246170TIFF2026016368000063.tif254170TIFF2026016368000064.tif254170TIFF2026016368000065.tif254170

[0336] Effects of compounds on CsgA aggregation, CsgA expression, and αSyn aggregation, i.e., white ThT uptake with 2 μM CsgA, 100 μM and 25 μM compounds in a color plate. Mean CsgA maximal aggregation (B) as a percentage of DMSO control in s max ) , median Abs IC in CsgA reporter assay 50 , and 50 μM αSyn and αSy compared to DMSO control in ThT assays using 100 μM compounds The mean inhibition rate of n aggregation was investigated.

[0337] [Table 4] TIFF2026016368000067.tif254170TIFF2026016368000068.tif254170TIFF2026016368000069.tif25417 0TIFF2026016368000070.tif254170TIFF2026016368000071.tif229170TIFF2026016368000072.tif81170

[0338] The % inhibition ranges presented are defined as follows: (--) is less than -10 (-) represents -10 to 10, (+) represents 11 to 30, and (++) represents 31. represents 60, and (+++) represents greater than 60.

[0339] Control B shown max The ranges for % and control AUC% are defined as follows: , (++++) represents 10 or less, (+++) represents 11 to 30, and (++) represents 31 or The numbers represent 1 to 60, and (+) represents greater than 60.

[0340] Presented reporter assay IC 50The range of (μM) is defined as follows: That is, (****) indicates less than 1.3, (***) indicates 1.3 to 4.5, and (** ) represents 4.6 to 10, and (*) represents greater than 10.

[0341] Effects of compounds on CsgA aggregation, CsgA expression, and αSyn aggregation, i.e., black ThT assay with 10 μM CsgA, 100 μM and 25 μM compounds in a color plate. Mean AUC as a percentage of DMSO control in the CsgA reporter assay Median Abs IC in assay 50 , and 50 μM αSyn and 100 μM compound Mean percent inhibition of αSyn aggregation compared to DMSO control in the ThT assay using the compounds I looked into it.

[0342] Thioflavin T assay results for compound effects on aggregation. Activity range of compounds is shown in Table 3. and are defined in Table 4. The compounds tested showed thiol activity in the inhibition of aggregation of α-synuclein and CsgA. The compounds showed a range of activity in the Rabin T assay. Inhibition of the various types of aggregation described in Tables 3 and 4 was achieved by the use of compounds of the invention (e.g., those shown in Table 1). compounds) inhibit α-synuclein aggregation, α-synuclein synthesis by CsgA or other microbial amyloids Seeding of α-synuclein aggregation and the role of microbial ami These compounds may be useful in preventing the formation of Parkinson's disease. These findings suggest that these compounds may be useful in preventing or treating cerebrospinal fluid disease and other alpha-synucleinopathies. Oral administration of these compounds reduces the abundance of amyloid-producing microorganisms. It is possible to achieve relatively high concentrations in the gastrointestinal tract, and the compound is These compounds may inhibit the dissemination of α-synuclein. Inhibition of aggregation may be beneficial on its own or may be beneficial in preventing microbial amyloid-seeding α-synuclein. This may have a synergistic effect with the inhibition of α-synuclein aggregation from the enteric nervous system to the central nervous system. Braak's hypothesis of prion-like transmission of “Exploring Braak's Hypothesis of Parkins on's Disease,”Front.Neurol.,13 February 2017), these inhibitory effects were associated with the inhibition of α-glucan in both the enteric and central nervous systems. -synuclein aggregates and may be useful in preventing their propagation, and further, orally administered compounds The substance is the continuous seeding of α-synuclein aggregates by microbial amyloid or α-synuclein. and removing α-synuclein aggregates if the subject mitigates the independent formation of α-synuclein aggregates. The process can have a greater net effect (i.e., This process can keep up with the formation of aggregates, thereby The product is intended to prevent or treat Parkinson's disease and other microbial amyloid-disseminated alpha-synucleinopathies. Without being limited by theory, oral administration may be effective in treating gastrointestinal It has been identified that the drug may restore gastrointestinal function in patients with gastrointestinal dysfunction. or preventing or delaying further loss of gastrointestinal function; and / or patients with alpha-synucleinopathy or those who have developed alpha-synucleinopathy For subjects at risk of developing dysphagia, reduced intestinal motility, gastrointestinal paresis, constipation (chronic Constipation (including chronic constipation and chronic idiopathic constipation), small intestinal bacterial overgrowth (SIBO), diarrhea, abdominal pain, and / or cramping, bloating, flatulence, nausea or any other symptoms of irritable bowel syndrome (IBS), Inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, intestinal hyperpermeability, or any of these may provide particular benefit in improving one or more symptoms of a combination of .

[0343] As shown in Tables 3 and 4, some compounds showed one or two titers in the assay. While the compounds showed inhibition of aggregation of only one type, other compounds were inactive or showed inhibition of aggregation of more than one type. Some compounds also appeared to enhance the aggregation of CsgA-seeded alpha-Sy n aggregation.

[0344] Example 27 In vitro live-cell fluorescent reporter assay to examine the effects of compounds on CsgA transcription To evaluate the effect of compounds on CsgA gene expression, the csgBAC promoter was used. Manipulation of Escherichia coli UTI89 with a fluorescent CsgA gene expression reporter driven by a chromatin A viable cell assay was performed using the constructed strain. Small-molecule inhibitors target Escheri chia coli amyloid biogenesis and biofilm formation, ´´Nat.Chem.Bio., 25 Oct 2009 In a reporter strain, a vector encoding green fluorescent protein (GFP) was used as described. The gene is inserted into the csgDE promoter, which contains the csgBAC promoter and other transcriptional regulatory elements. The complete intergenic region between F and csgBAC was placed immediately downstream of the E. coli UTI89 staining A single copy was inserted into the chromosome. With this construct, GFP expression in the reporter strain was regulated by Cs The reporter strain was plated on YESCA agar plates (1 g / L of yeast Extract, 10g / L Casamino Acids, 20g / L Bacto Agar , and grow at approximately 27°C for 2 days or until single colonies are large enough to pick. A single colony of the reporter strain was used to grow the 5 mL of YESCA medium (1 g / L yeast The inoculation was carried out on a medium containing 10 g / L of the parent extract (cassamino acids) and grown overnight at approximately 27°C. The seed material was diluted 1:10 in YESCA medium and transferred to a SpectraMax™ M 2 or M5e (Molecular Devices, San Jose, CA) The optical density (OD) at 600 nm was measured using a spectrophotometer. 600 The culture was then further Dilute to a final OD of 0.003 600 This is achieved by OD 600 Confirmed by measuring The compounds were diluted in dimethyl sulfoxide (DMSO) to final test concentrations of 0.14 to 100 μM. Dilute to a concentration of 1.5 μL per well in a black, clear-bottom, tissue-culture-treated 96-well plate. All assays, including vehicle controls and positive controls for inhibition, were plated on a single plate. Assay wells contained a final 1% DMSO. Vehicle control wells contained no To assess reporter strain expression in the assay in the absence of the compound, DMSO and Positive controls for inhibition consisted of Csg in WT E. coli. Compound 020, which was previously observed by qRT-PCR to inhibit A expression, and a final concentration of 10 g / L of salt that largely or completely inhibits CsgA expression in vitro. Wells containing OD in YESCA medium were included. 600 0.003 of 148.5 μL Culture medium was added to all wells to a final volume of 150 μL. The mixture was incubated at 7°C for 20 hours, and then the GFP fluorescent signal in each well was measured at 485n The SpectraM The OD of each well was measured using a x M2 or M5e plate reader. 600 Spect The measurements were performed using a ramax M2 or M5e plate reader. Since CsgA expression was completely inhibited under the growth conditions of YE containing 10 g / L NaCl, Background fluorescence was calculated by averaging the values ​​from wells containing bacteria during SCA. After background fluorescence was subtracted from all wells, the fluorescence signal of each well was calculated. The normalized dose-response curves were plotted against the mean of the vehicle control wells. Compound inhibition of CsgA transcription was plotted using GraphPad Prism. Software version 8.1.1 (GraphPad Software, San Diego) (Gibberish, CA) to curve fit IC 50 , bottom, top, span( The results were analyzed by assessing the top-bottom and Hill slope. An additional metric used was the concentration at which the curve reached 50% of the expression level of the vehicle control. This is the X value, i.e., the fluorescence of the reporter strain, in the formula X

[50] , or Y=50. The compound concentration calculated to reach 50% of vehicle control fluorescence was calculated according to GraphPad. This was described as a custom analysis in Prism. Apart from the fluorescence data, OD 600 value The values ​​were normalized to the mean of the vehicle control wells and dose-response curves were plotted for each compound. More than 10% OD 600 The decrease in β indicates that the compound has some toxicity to bacterial growth. or have been shown to have adverse effects.

[0345] In some embodiments, other versions of live cell reporter assays for CsgA expression are In this case, a plasmid-encoded reporter construct can be used. The E. coli intergenic region containing the AC promoter and all transcriptional regulatory elements is located immediately behind The same reporter, consisting of the GFP gene, is driven by a constitutive promoter. The plasmid may contain a selectable marker such as a resistance gene. The plasmid-borne reporter may be expres...

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof: 【Chemistry 1】 A １ is -C(R ７ ) (R ８ ) or —CO—, and X is —SO ２ -, CO or -C( R ９ ) (R １０ ) if A １ HA-N(R ７ ) - and A ２ is absent or -C(R ５ ) (R ６ ) and L １ is a bond, (-CH ２ -) ｍ , -CF ２ -, -(C=O)- or -C(R ９ ) (R １０ ) - and L ２ is a bond, (-CH ２ -) ｍ , -CF ２ -, -(C=O)- or -C(R ９ ) (R １０ ) - and EO、-N(R １１ )-、-N(R １４ )-、-O-、-C-、-S-、-S(=O ) -, -SO ２ -, -CF ２ -, -C(R ９ ) (R １０ ) - and Y is O or S; Z is =C(R １３ )-, ═N-, or -N(R １１ ) - and R １ is substituted or unsubstituted phenyl, substituted or unsubstituted heterocyclyl, R ２ is a substituted or unsubstituted naphthyl, a substituted or unsubstituted phenyl, a substituted or unsubstituted Substituted quinolinyl, substituted or unsubstituted isoquinolinyl, or substituted or unsubstituted heteroquinolinyl It is a krill, R ３ and R ４ is independently absent, or is -H, substituted or -, where valence permits. unsubstituted C １ ~C １０ Alkyl, acyl, —CO ２ R ７ , -CON(R ７ ) (R ８ ), -P =O(OH) ２ or -SO ２ (OH), R ５ and R ６ are independently absent or -H and C, as far as valence permits. １ ~C １ ０ alkyl, or in combination with each other to form a spirocarbocyclic ring or spiro(heterocyclic ring). (hetero) carbocyclic rings, R ７ and R ８ is independently absent, or is -H, substituted or -, where valence permits. unsubstituted C １ ~C １０ Alkyl, -(CH ２ ) ｍ -aryl, -(CH ２ ) ｍ - Hetero ant , -(CH ２ ) ｍ -substituted or unsubstituted cycloalkyl, -R １４ Selected from combine to form a spiropentanyl ring, R ９ and R １０ is independently, at each occurrence, —H, —Cl, —Br, —F, —CF ３ , C １ ~C １０ is alkyl, R １１ and R １２ is independently, at each occurrence, —H, acyl, sulfonyl, substituted or unsubstituted; or unsubstituted C １ ~C １０ Alkyl, C ３ ~C ６ Cycloalkyl, C ３～６ Heterocyclyl , substituted or unsubstituted benzyl, -(CH ２ ) ｏ -(substituted or unsubstituted aryl) or - (CH ２ ) ｏ -(substituted or unsubstituted heteroaryl), R １３ は、-H、-OH、-OR １１ 、-Cl、-Br、-F、-CN、-CF ３ 、- CH ２ F, -CHF ２ , substituted or unsubstituted C １ ~C １０ Alkyl, C １ ~C １０ Alkenyl , C ３ ~C ６ Cycloalkyl, substituted or unsubstituted C ３ ~C ６ Heterocyclyl, acyl, -C O ２ R ７ 、-(CH ２ ) ｍ CO ２ N(R １１ )(R １２ )、-CON(R ７ )(R ８ )、 -(CH) ２ ) ｍ OH, -(CH) ２ ) ｍ CO ２ H, -(CH) ２ ) ｍ NH ２ 、-(CH ２ ) ｍ N(R １１ )(R １２ )、-N(R １１ )(R １２ )、-NR １１ (C=O)(CH ２ ) ｍ CH ３ , -NR １１ (C=O)R １２ and NR １２ (SO ２ ) (CH ２ ) ｍ CH ３ from Selected, R １４ is -H, C １ ~C １０ Alkyl, C １ ~C １０ Alkenyl, C １ ~C １０ (Mo mono- or polyhydroxylated alkyl, -(CH ２ ) ｏ -R １５ , -(CH ２ CH ２ O) ｏ -R １５ 、-(CH ２ ) ｍ -CO ２ H, -(CH) ２ ) ｍ -NH ２ 、-(CH ２ ) ｍ --- CO)NR １６ R １７ or a protecting group, R １５ is -CON(R １１ ) (R １２ ), -N(R １１ ) (R １２ ), acyl, -C O ２ R ７ , substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cyclohexyl chloroalkyl, substituted or unsubstituted heterocyclyl; R １６ and R １７ are independently —H or —CH ３ and m is, independently in each occurrence, 0 to 10; n is independently 1 to 5 in each occurrence; o is independently, at each occurrence, 1 to 20; 【Chemistry 2】 represents a single bond or a double bond, However, the compound of formula (I) 【Transformation 3】 or a pharmaceutically acceptable salt thereof.

2. X is -SO ２ The compound of claim 1, wherein

3. A １ Ga-NR １４ The compound of claim 1, wherein

4. A １ -C(R ７ ) (R ８ 2. The compound of claim 1, wherein

5. A ２ -C(R ５ ) (R ６ 2. The compound of claim 1, wherein

6. A ２ The compound of claim 1 , wherein

7. L １ The compound of claim 1 , wherein is a bond.

8. L ２ -C(R ９ ) (R １０ 2. The compound of claim 1, wherein

9. The compound of claim 1 , wherein Y is O.

10. The compound of claim 1 wherein Y is S.

11. Z = C(R １３ 2. The compound of claim 1, wherein

12. Z is =N- or -N(R １１ 2. The compound of claim 1, wherein

13. Z is -N(R １１ 2. The compound of claim 1, wherein

14. R １ The compound of claim 1 , wherein is trifluoromethylphenyl.

15. R ２ The compound of claim 1 , wherein is unsubstituted naphthyl.

16. R ３ is -H, and R ４ Ga-CO ２ R ７ 2. The compound of claim 1, wherein:

17. Formula (II) or Formula (III): 【Chemistry 4】 2. The compound of claim 1 having the structure:

18. Formula (IIa), Formula (IIb) or Formula (IIc): 【Transformation 5】 having the structure During the ceremony, L １ is a bond, (-CH ２ -) ｍ , -CF ２ - or -(C═O)-, L ２ is a bond, (-CH ２ -) ｍ , -CF ２ - or -(C═O)-, m is, independently in each occurrence, 0 to 10; R １ is substituted or unsubstituted phenyl, substituted or unsubstituted heterocyclyl, R ２ is a substituted or unsubstituted naphthyl or a substituted or unsubstituted heterocyclyl 、 R ３ は、-CO ２ H、 【Transformation 6】 and R ５ and R ６ are H, respectively, R ７ is a substituted or unsubstituted C １ ~C １０ Alkyl, or substituted or unsubstituted cycloalkenyl It is Lukil, R １３ is -H, -Cl, -Br, -F, -CN, -CF ３ , -CH ２ F, -CHF ２ , substituted or unsubstituted C １ ~C １０ Alkyl, —NH ２ , -CONH ２ , -(CH ２ )-N( CH ３ ) ２ , -NH(cyclopentyl), -NH(benzyl), -NH(tetrahydropyridine) Ran), -NH-(CH ２ ) (cyclopentyl) and —O—(CH ２ ) ２ -From phenyl 18. The compound of claim 17, wherein

19. Compound 001, Compound 002, Compound 005, Compound 006, Compound 015, Compound 0 15a, compound 015b, compound 016, compound 016a, compound 016b, compound 01 7, Compound 018, Compound 021, Compound 021a, Compound 021b, Compound 024, Chemical Compound 025, Compound 025a, Compound 025b, Compound 026a, Compound 026b, Compound compound 028, compound 029, compound 030, compound 031, compound 032, compound 033, Compound 034, Compound 035, Compound 036, Compound 037, Compound 038, Compound 03 9, Compound 040, Compound 041, Compound 042, Compound 043, Compound 044, Compound 045, Compound 046, Compound 047, Compound 048, Compound 049, Compound 052, Chemical Compound 056, Compound 058, Compound 065a, Compound 065b, Compound 066, Compound 0 67, compound 068, compound 070, compound 071, compound 072, compound 074, compound compound 076, compound 077, compound 078, compound 079, compound 082, compound 084, Compound 085, Compound 086, Compound 087, Compound 088, Compound 089, Compound 09 0, compound 091, compound 092, compound 093, compound 094, compound 095, compound 096, Compound 097, Compound 098, Compound 099, Compound 100, Compound 101, Chemical Compound 102, Compound 105, Compound 106, Compound 107, Compound 108, Compound 109 , Compound 116, Compound 117, Compound 120, Compound 121, Compound 122, Compound 1 23, compound 124, compound 125, compound 126, compound 127, compound 128, compound compound 131, compound 132, compound 137, compound 138, compound 141, compound 142, Compound 143, Compound 144, Compound 145, Compound 146, Compound 147, Compound 14 8, Compound 149, Compound 150, Compound 151, Compound 152, Compound 153, and 18. The compound of claim 17, selected from the pharmaceutically acceptable salts thereof:

20. Compound 112, Compound 113, Compound 114, Compound 115, Compound 139, Compound 1 54, compound 156, compound 157, compound 158, compound 159, compound 160, compound compound 172, compound 173, compound 175, compound 176, compound 177, compound 183, Compound 184, Compound 186, and pharmaceutically acceptable salts thereof. Item 18. The compound according to item 17.

21. Formula (IV), Formula (V), Formula (VI), Formula (VII), Formula (VIII), Formula (IX), or is the formula (X): 【Transformation 7】 2. The compound of claim 1 having the structure:

22. Formula (IXa), Formula (IXb) or Formula (IXc): 【Transformation 8】 having the structure During the ceremony, L １ is a bond, (-CH ２ -) ｍ , -CF ２ - or -(C═O)-, L ２ is a bond, (-CH ２ -) ｍ , -CF ２ - or -(C═O)-, m is, independently in each occurrence, 0 to 10; o is, independently at each occurrence, 1 to 20; R １ is substituted or unsubstituted phenyl, substituted or unsubstituted heterocyclyl, R ２ represents substituted or unsubstituted naphthyl, substituted or unsubstituted quinolinyl, substituted or unsubstituted isoquinolinyl, substituted phenyl, or substituted or unsubstituted heterocyclyl; R ３ は、-CO ２ H、 【Chemistry 9】 and R １３ is -H, -Cl, -Br, -F, -CN, -CF ３ , -CH ２ F, -CHF ２ , -(CH ２ ) ｍ -NMe ２ , -CONH ２ , -CO ２ H, and substituted or unsubstituted C １ ~C １０ alkyl, R １４ is C １ ~C １０ Alkyl, -(CH ２ ) ｏ -(unsubstituted cycloalkyl), -( CH ２ ) ｏ -(substituted or unsubstituted phenyl), -(CH ２ ) ｏ -naphthyl or -(CH ２ ) ｏ -biaryl.

23. Compound 004, Compound 004a, Compound 004b, Compound 014, Compound 014a, Compound 014b, Compound 019, Compound 022, Compound 027a, Compound 027b, Compound 050, Compound 053, Compound 054, Compound 055, Compound 057, Compound 059, Chemical Compound 060, Compound 061, Compound 062, Compound 063, Compound 064, Compound 069 , compound 073, compound 075, compound 080, compound 081, compound 083, compound 1 03, compound 104, compound 110, compound 111, compound 112, compound 113, compound compound 114, compound 115, compound 118, compound 129, compound 130, compound 134, Compound 135, Compound 136, Compound 140, and pharmaceutically acceptable salts thereof.

22. The compound of claim 21 , wherein

24. Compound 003, Compound 023, Compound 051, Compound 119, Compound 133, Compound 1 51, Compound 155, Compound 162, Compound 163, Compound 164, Compound 165, Compound compound 166, compound 167, compound 168, compound 169, compound 170, compound 171, Compound 174, Compound 178, Compound 179, Compound 180, Compound 181, Compound 18 2, Compound 187, Compound 189, Compound 190, Compound 191, Compound 192, Compound 194, Compound 195, Compound 196, Compound 197, Compound 198, Compound 199, Chemical Compound 200, Compound 201, Compound 202, Compound 203, Compound 204, Compound 205 , compound 206, compound 207, compound 208, compound 209, compound 210, compound 2 11, Compound 212, Compound 213, Compound 214, Compound 215, Compound 216, Compound compound 217, compound 218, compound 219, compound 220, compound 221, compound 222, Compound 223, Compound 224, Compound 225, Compound 226, Compound 227, Compound 22 8, Compound 229, Compound 230, Compound 231, Compound 232, Compound 233, Compound 234, Compound 235, Compound 236, Compound 237, Compound 238, Compound 239, Chemical Compound 240, and pharmaceutically acceptable salts thereof. Compound.

25. A compound according to any one of claims 1 to 24 or a pharmaceutically acceptable salt thereof and a pharmaceutical A pharmaceutical composition comprising a physiologically acceptable carrier.

26. 26. The medicament of claim 25, wherein the compound or the pharmaceutical composition is formulated for oral administration. composition.

27. 26. The compound or pharmaceutical composition of claim 25, wherein the compound or pharmaceutical composition is formulated for administration by injection. A pharmaceutical composition comprising:

28. 28. The method of claim 27, wherein the injection is intravenous, subcutaneous, intramuscular, intraperitoneal, intraspinal, or intracranial. The pharmaceutical composition described.

29. 26. The pharmaceutical composition of claim 25, formulated for enteral delivery.

30. 26. The pharmaceutical composition of claim 25, formulated for controlled release in the lower intestinal tract or colon of a subject. Finished product.

31. 31. The pharmaceutical composition of claim 30, further comprising a formulation for enteral delivery.

32. 26. The pharmaceutical composition of claim 25, formulated for delivery to the central nervous system of a subject.

33. 26. The pharmaceutical composition of claim 25, formulated for delivery outside the systemic circulation of a subject.

34. 34. The pharmaceutical composition of claim 33, formulated for topical delivery.

35. 34. The pharmaceutical composition of claim 33, formulated for mucosal delivery.

36. 32. The pharmaceutical composition of claim 31, formulated for delivery to the oral cavity.

37. Methods for inhibiting amyloid formation in a subject in need thereof The compound according to any one of claims 1 to 24 or a pharmaceutically acceptable salt thereof. or the pharmaceutical composition according to any one of claims 25 to 36 to the subject. and

38. Preventing or treating an amyloid disorder in a subject in need thereof. A method for treating a patient suffering from atopic dermatitis, comprising administering to said patient a compound according to any one of claims 1 to 24 or a pharmaceutical composition thereof. or the pharmaceutical composition according to any one of claims 25 to 36. Administering

39. 39. The method of claim 38, wherein the amyloid disorder is a neurological disorder.

40. The amyloid disorder is Parkinson's disease (PD), dementia with Lewy bodies, or multiple system atrophy. , multiple sclerosis (MS), frontotemporal dementia (FTD), REM sleep behavior disorder (RBD), α-synucleinopathy, PD-related constipation, PD-related hypotension, Huntington's disease, Alexa 38. The method according to claim 38, wherein the disease is von Bender's disease, amyotrophic lateral sclerosis (ALS), or Alzheimer's disease. The method described.

41. The amyloid disorder is intestinal dysbiosis, intestinal hyperpermeability, irritable bowel syndrome (IB), S), inflammatory bowel disease (IBD), ulcerative colitis or Crohn's disease. How to do it.

42. The subject is a patient suffering from dysphagia, decreased intestinal motility, gastroparesis, constipation (chronic constipation and chronic idiopathic constipation), (including constipation), small intestinal bacterial overgrowth (SIBO), diarrhea, abdominal pain and / or cramps, bloating , flatulence, hypersalivation (excessive salivation), anorectal dysfunction, bowel coordination disorder, and nausea 39. The method of claim 38, wherein the patient is suffering from a gastrointestinal condition comprising one or more of:

43. The gastrointestinal symptoms are caused by alpha-synucleinopathy, Parkinson's disease or parkinsonism.

43. The method of claim 42, wherein the method is associated with a system.

44. The amyloid disorder may be diagnosed by detecting the presence or level of gut bacterial amyloid protein. The method according to any one of claims 38 to 43, wherein the diagnosis can be made by:

45. 45. The method of claim 44, wherein the protein comprises a bacterial CsgA protein.

46. A bacterial protein, such as CsgA, or a bacterial protein-producing bacterial protein, in an intestinal sample of the subject.

46. ​​The method of claim 38, further comprising detecting the presence or level of a viable microorganism.

1. The method according to claim 1.

47. In the intestinal sample, the bacterial protein, the transcript mRNA of the bacterial protein, or if the presence of said microorganism producing said bacterial protein is detected, or if said intestinal sample is wherein the level of the bacterial protein or the microorganism producing the bacterial protein is If the level is greater than a predetermined level or a control, the subject is considered to be in need of the prevention or treatment.

47. The method of claim 46, wherein

48. 48. The method of claim 47, wherein said detecting is by qPCR or RT-qPCR.

49. and further comprising confirming the reduction or absence of intestinal amyloid protein after administration. Item 49. The method according to any one of Items 43 to 48.

50. 49. The method of claim 43, further comprising identifying the subject as having a gastrointestinal symptom.

10. The method according to any one of the preceding claims.

51. The compound or precursor of amyloid according to any one of claims 1 to 24 and a pharmaceutically acceptable salt thereof. Methods for preventing and / or inhibiting the formation of

52. In a subject in need of preventing the formation of amyloid protein aggregates, 1. A method for disrupting protein aggregate formation, comprising: A compound according to any one of claims 1 to 24 or a pharmaceutically acceptable salt thereof, or comprising administering to said subject a pharmaceutical composition according to any one of claims 25 to 33. ,method.

53. Before administering the compound or the pharmaceutical composition, the formation of amyloid protein aggregates is inhibited. selecting subjects to benefit from compounds that interfere with the and measuring the inhibition of the formation of aggregates of the amyloid protein in the presence of the α-amyloid protein.

52. The method according to claim 52.

54. 54. The method of claim 53, wherein said selecting is by clinical or diagnostic evaluation. 。

55. 54. The amyloid protein of claim 51, wherein the amyloid protein comprises one or more mammalian proteins. The method according to any one of the preceding claims.

56. The mammalian protein is derived from α-synuclein, tau, amyloid precursor protein Beta-amyloid, medin, apolipoprotein AI, atrial natriuretic factor, Beta-amyloid, cystatin, IAPP (amylin), beta-2 microglobulin, Transthyretin, PrP, gelsolin, lysozyme, huntingtin, keratoepithelial Keratoepithelin, calcitonin, prolactin, serum amyloid A, SOD1 and immunoglobulin light chain AL.

57. the amyloid protein comprises one or more bacterial or fungal proteins; 57. The method of any one of claims 51 to 56.

58. 58. The method of claim 57, wherein the bacterial protein is CsgA or a CsgA homologue. 。

59. The CsgA homologue is Citrobacter farmeri farmeri), Salmonella enterica ), Enterobacter cloacae, Bacillus velezensis, Pseudomonas Pseudomonas stutzeri, Burkholderia cepa Burkholderia cepacia, Hafnia albaii one or more selected from Pseudomonas alvei and Pseudomonas reinekei 59. The method of claim 58, wherein the organism is produced by the above organism.

60. The amyloid protein is present in the gastrointestinal tract, the dural sinuses, the oral cavity, or the nasal cavity.

60. The method of any one of claims 51 to 59.

61. 61. The method of claim 51, wherein the amyloid protein is present in enteric nerve tissue or the olfactory bulb.

10. The method according to any one of claims 1 to 9.

62. Measure enteric amyloid levels and / or amyloid protein during the administration process.

62. The method of any one of claims 52 to 61, further comprising:

63. After the administration course, the level of enteric amyloid and / or amyloid protein is measured or 63. The method of any one of claims 52 to 62, further comprising evaluating

64. and further measuring or assessing changes in the nervous system, such as neurological symptoms or behavior of the subject.

64. The method of any one of claims 52 to 63, comprising:

65. and measuring or assessing changes in the gastrointestinal system, such as gastrointestinal symptoms or behavior, of said subject. The method of any one of claims 52 to 64.

66. 66. The method of claim 65, wherein the gastrointestinal symptom comprises constipation.

67. wherein the compound or the pharmaceutical composition is administered before the onset of a neurological symptom or condition.

67. The method of any one of claims 52 to 66.

68. The neurological symptom or condition is anosmia, dysosmia, bradykinesia, ataxia, tremor, muscle rigidity Shrinkage, posture and balance disorders, loss of motor activity, dysarthria or other changes in speech, changes in handwriting , orthostatic hypotension, memory impairment, swallowing disorders, incontinence, sleep disorders, cardiac arrhythmias, visual disturbances, depression and Psychiatric problems including visual, auditory, olfactory or tactile hallucinations, dizziness, cognitive impairment, changes in vasopressin levels, changes in serotonin levels, changes in kynurenine levels, and / or 68. The method of claim 67, comprising one or more of any combination of these.

69. The compound or the pharmaceutical composition is selected from the group consisting of caffeine, nicotine, theophylline, theobroma, 52. The method of claim 52, wherein the compound is co-administered with a benzodiazepine, a xanthine, a methylxanthine, or a derivative thereof.

69. The method of any one of claims 1 to 68.

70. Claims 52 to 56 further comprising administering to the subject an α-synuclein aggregation inhibitor.

10. The method according to any one of claims 9 to 10.

71. The subject is at risk of developing Parkinson's disease, such as by clinical or diagnostic assessment. Any of claims 52 to 70, wherein the subject is identified or selected as having or already having 10. The method according to claim 1.

72. The subject is diagnosed, e.g., by clinical or diagnostic assessment, with dementia with Lewy bodies, concomitant Lewy bodies, -body disease, Lewy body variant of Alzheimer's disease, multiple system atrophy, pure autonomic failure identified as at risk of developing or already having, or any combination thereof The method of any one of claims 52 to 71, wherein the selected

73. A method for preventing or treating an inflammatory disorder in a subject in need thereof A method for treating a rheumatoid arthritis, comprising administering to a patient a compound according to any one of claims 1 to 24 or a pharmaceutically acceptable salt thereof. or a pharmaceutical composition according to any one of claims 25 to 33 is administered to the subject. A method comprising:

74. The disorder may be bacterial sepsis, autoimmune disease, lupus erythematosus, ischemia-reperfusion injury, or cerebral 74. The inflammation of claim 73, selected from stroke, metabolic disease, obesity-related metabolic inflammation, gout, and cancer. Idiopathic disorder.

75. 75. The inflammatory disorder of claim 74, wherein the disorder is lupus erythematosus.

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