Relatively insufficient-supply amyloid beta aggregation inhibitors for improving detoxification effects

Abstract

Provided herein is a method of treating or preventing protein misfolding and deposition diseases in a subject by administering to the subject a low dose of Compound (1). Also provided herein is a method of reversing or preventing the toxic effect of the misfolded and aggregated protein amyloid beta by using a relatively insufficient supply (reverse stoichiometric) compound (1) to achieve a stronger detoxification effect. The invention further provides the dosage and the application mode of the composition.

Description

Background Art

[0001] Most protein molecules must fold into a defined three-dimensional structure to acquire functional activity. However, protein chains can adopt multiple conformational states, and their biologically active conformations are often only marginally stable. Proteins that are in a conformational stage that does not support their physiological function are said to be misfolded. Aggregation and accumulation of misfolded proteins may lead to disease, a phenomenon that increases dramatically with age. A mechanistic explanation for this association is that as humans age (or as a result of mutations), the delicate balance of protein synthesis, folding, and degradation may be disturbed, leading to the generation and accumulation of misfolded proteins, forming aggregates and inducing pathology. Protein aggregation diseases are not limited to the central nervous system; they may also occur in peripheral tissues.

[0002] One of the most common protein misfolding diseases is Alzheimer's disease, which is associated with the misfolding of the protein amyloid β. Misfolded amyloid β shows a tendency to aggregate and form oligomeric species, which are known to be highly toxic to neuronal cells, leading to neurodegeneration.

[0003] This amyloid-β-related pathology is not limited to different forms of Alzheimer's disease, it is also present in type II diabetes, serum amyloid A disease (SAA amyloidosis), hereditary Iceland syndrome, multiple myeloma, medullary carcinoma, aortic amyloidosis, cardiac amyloidosis, insulin injection amyloidosis, prion systemic amyloidosis, chronic inflammatory amyloidosis, senile systemic amyloidosis, pituitary amyloidosis, hereditary renal amyloidosis, familial British dementia, Finnish hereditary amyloidosis, familial non-neurological amyloidosis, amyloid-β-related eye diseases, especially glaucoma and age-related macular degeneration, etc.

[0004] The formation of pathological aggregates of misfolded proteins has been identified as a promising target for new therapeutic approaches that have the potential to slow down or even stop the progression of neurodegenerative processes. Just recently, especially soluble toxic amyloid-β oligomers have been shown to be a promising drug target for Alzheimer's disease, with the first new treatments being successfully developed in clinical trials and two recently approved Alzheimer's drugs (aducanumab and lecanemab) being approved for treatment.

[0005] Despite these first successful new drug developments using antibody-based infusions to target toxic amyloid-β oligomers and fibrils, their beneficial effects on Alzheimer's disease progression remain rather limited, and there remains a great need for more effective, better tolerated, easier to administer, and cheaper to produce drugs. Summary of the invention

[0006] In some aspects, disclosed herein is a method of treating or preventing a protein misfolding and deposition disease in a subject in need thereof, the method comprising administering to the subject a low dose of compound (1).

[0007]

[0008] Compound (1),

[0009] Thereby treating or preventing a protein misfolding and deposition disease in said subject.

[0010] In some related aspects, the low dose administration results in a concentration at the site of action in the brain of less than 50 nM. In some related aspects, the low dose administration results in a concentration at the site of action of 10 nM. In some related aspects, the low dose administration results in a concentration at the site of action of 3 nM. In some related aspects, the low dose administration results in a concentration at the site of action of 1 nM.

[0011] In some related aspects, the protein misfolding and deposition disease is selected from a disease associated with amyloid beta pathology, i.e., Alzheimer's disease (AD), early-onset Alzheimer's disease, late-onset Alzheimer's disease, presymptomatic Alzheimer's disease, type II diabetes, serum amyloid A disease (SAA amyloidosis), hereditary Iceland syndrome, multiple myeloma, medullary carcinoma, aortic amyloidosis, cardiac amyloidosis, insulin injection amyloidosis, prion systemic amyloidosis, chronic inflammatory amyloidosis, senile systemic amyloidosis, pituitary amyloidosis, hereditary renal amyloidosis and familial non-neurological amyloidosis.

[0012] In some related aspects, protein misfolding and deposition diseases include amyloid-related diseases. On the one hand, amyloid-related diseases include Alzheimer's disease (AD), early-onset Alzheimer's disease, late-onset Alzheimer's disease, presymptomatic Alzheimer's disease or any combination thereof. On the one hand, amyloid-related diseases are Alzheimer's disease (AD).

[0013] In some related aspects, the protein misfolding and deposition disease is selected from, but not limited to, Parkinson's disease and related synucleinopathies, Huntington's disease, prion diseases, mad cow disease, Creutzfeldt-Jakob disease, spongiform encephalopathies, frontotemporal dementia spectrum disorders, amyotrophic lateral sclerosis, familial British and Danish dementia, Finnish hereditary amyloidosis, familial non-neural amyloidosis, and retinitis pigmentosa.

[0014] In some related aspects, administration of compound (1) is systemic, using oral administration, rectal administration, transmucosal administration, intranasal administration, intramuscular administration, subcutaneous administration, transdermal administration, intrathecal administration, direct intracerebroventricular administration, intravenous administration, intraperitoneal administration, or intranasal instillation.

[0015] In some related aspects, the administration is oral administration.

[0016] In some further related aspects, the oral administration comprises a dose of compound (1) in the range of 0.03-0.3 mg / kg.

[0017] In some further related aspects, the intravenous administration comprises a dose of compound (1) in the range of 0.01-.1 mg / kg.

[0018] In some other aspects, amyloid protein related diseases are selected from neurodegenerative retinal diseases, particularly glaucoma or age-related macular degeneration (AMD). In some aspects, glaucoma is selected from primary angle-closure glaucoma, open-angle glaucoma (primary and secondary), wide-angle glaucoma, steroid-induced glaucoma, traumatic glaucoma, secondary angle-closure glaucoma and neovascular glaucoma. In some aspects, age-related macular degeneration (AMD) is selected from dry type and wet type AMD, also includes the geographic atrophy (GA) secondary to AMD.

[0019] In some further aspects, the amyloid-related disease is selected from diabetic retinopathy. In some further aspects, the diabetic retinopathy is non-proliferative diabetic retinopathy, proliferative diabetic retinopathy, or a combination thereof.

[0020] In some aspects, the administration is ocular administration.

[0021] In some aspects, ocular administration is by eye drops, eye creams, eye ointments, eye sprays, intraocular depot formulations, by injection including intraocular injections, intraocular or periocular injections including intraocular fluids, depot formulations, solid and semisolid intraocular vehicles and matrices, or ophthalmic devices such as contact lenses and membranes.

[0022] In some aspects, topical ocular administration comprises eye drops containing less than 1 mg of compound (1) per drop. In some further aspects, ocular administration comprises a dose of less than 3 mg of compound (1) per drop.

[0023] In some aspects, the method comprises administering to the subject an initial loading dose of compound (1), and further administering a plurality of subsequent maintenance doses of the compound. In some aspects, the subsequent maintenance doses are lower doses of compound (1).

[0024] In some aspects, the compound is administered to the subject daily for a first period of at least one day, followed by a second period of at least one week in which the compound is not administered, followed by a repetition of the at least one day first period in which the subject is administered the compound every day.

[0025] In some aspects, compound (1) is the active ingredient of a pharmaceutical composition, which further comprises a physiologically acceptable carrier. In some other aspects, the pharmaceutical composition is suitable for oral administration. In some other aspects, the pharmaceutical composition is suitable for ocular administration.

[0026] In some aspects, disclosed herein is a method of inhibiting amyloid βAβ toxicity in a subject, the method comprising administering a low dose of compound (1)

[0027]

[0028] Compound (1),

[0029] thereby inhibiting Aβ in the subject 1-42 toxicity.

[0030] In some aspects, disclosed herein is a method for reversing or preventing the toxic effects of misfolded and aggregated protein amyloid β, i.e., toxic Aβ oligomers, by using a relatively insufficient supply (reverse stoichiometric ratio) of compound (1) to achieve a stronger detoxification effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] The subject matter which is regarded as the invention is particularly pointed out and distinctly claimed in the concluding portion of the specification. However, the invention, both as to its organization and method of operation, together with objects, features, and advantages thereof, may be best understood by reference to the following detailed description when read in connection with the accompanying drawings in which:

[0032] Figure 1 Compound (1) shows the effect of surface plasmon resonance (SPR) on amyloid βAβ 1-42 Graphical illustration of the target affinity of monomers. 1-42 For this binding assay, the assay was performed using a Biacore X100 biosensor instrument equipped with two flow cells. Aβ was coupled to 1-42 A flow cell covalently coupled to a CM7 sensor chip. HBS-EP was used as the assay running buffer. Analyte compound (1) with a concentration range of 0.3 nM to 1000 nM was injected onto the sensor chip at a flow rate of 10 μL / min at 25°C for 180 seconds. The response was evaluated at steady state, plotted against concentration, and fitted to K by a four-parameter logistic equation. D=2.5nM (n=4). K D (dissociation constant) is a measure of affinity for the target and represents the concentration of compound (1) that induces half-maximal binding.

[0033] Figure 2 A graphical illustration showing the pharmacokinetics of compound (1) in rat brain. Compound (1) was administered as a subcutaneous (sc) injection at doses of 0.4 mg / kg (▽) and 2.0 kg / kg (O, △). A microdialysis probe was placed in the prefrontal cortex of these adult male Sprague Dawley rats to measure the concentration of compound (1) in the brain interstitial fluid (ISF) using an HPLC system with an MS / MS detector. Dialysate samples were collected at a flow rate of 0.10 μL / min and a vehicle flow rate of 0.8 μL / min. Animals were either anesthetized or awake and free-moving. Microdialysis data over a period of 360 min are plotted as the mean ± SEM of 4 measurements on a linear scale.

[0034] Figure 3 A graphical illustration showing the dose-dependent effects of compound (1) at very low concentrations on LTP in anesthetized rats. Normalized LTP of population peak (PS) amplitude at 80-90 min after theta burst stimulation (TBS) is shown. Compound (1) was administered sc at doses of 0.08 mg / kg, 0.4 mg / kg and 2.0 mg / kg. Amyloid βAβ was added at concentrations that strongly inhibited LTP. 1-42 injection into the cerebral ventricle (black bars). A 2 mg / kg sc dose of compound (1) (dark grey) normalized the LTP signal as expected, while reaching a brain concentration of approximately 60 nM (see Figure 2 ). However, the presumed inactive dose was only 0.4 mg / kg (medium grey), resulting in a peak brain concentration of 10 nM (see Figure 2 ), showed at least as effective efficacy against LTP. A further 5-fold reduction in the dose (light grey) resulted in loss of pharmacological effect.

[0035] Figure 4A -B. Figure 4A Shown is a graph from an in vitro LTP experiment using rat brain slices incubated with 50 nM Aβ at 23°C. 1-42 and / or 10 nM compound (1) for superperfusion. Compound (1) solution was added 90 min before HFS was delivered in the first infusion (dark grey circles). At this concentration, compound (1) allowed the induction of LTP. In order to ensure that Aβ 1-42 To determine the validity of the Aβ-mediated effect, we also performed Aβ 1-42 Crossover experiment (50 nM, closed circle, n = 12).1-42 In the presence of , 10 nM of compound (1) was able to prevent LTP blockade when LTP was induced in the second input (light grey circles). Representative field excitatory postsynaptic potentials (fEPSPs) are shown at the top.

[0036] Figure 4B Indicates based on Figure 4A LTP experiment shown, graphical illustration of the dose-dependent effect of compound (1) on LTP in vitro in rat brain slices. 1-42 Toxic Aβ monomers 1-42 The oligomers almost completely blocked the LTP response (black bars). Compound (1) at 500 nM and 100 nM only partially alleviated the Aβ 1-42 However, a 10-fold lower concentration of 10 nM had a significantly stronger effect and almost completely blocked Aβ 1-42 Surprisingly, the efficacy at 10 nM means that 1 molecule of compound (1) can detoxify 5 Aβ 1-42 This refutes the previous state of the science which required an excess of antidotes. Data are shown as mean ± SD of 5 experiments (n=5).

[0037] In addition, it should be understood that for simplicity and clarity of illustration, the elements shown in the figures are not necessarily drawn to scale. For example, for clarity, the dimensions of some elements may be exaggerated relative to other elements. In addition, reference numerals may be repeated between the figures to indicate corresponding or similar elements where deemed appropriate. DETAILED DESCRIPTION

[0038] In the following detailed description, a number of specific details are set forth to provide a thorough understanding of the present invention. However, those skilled in the art will appreciate that the present invention may be practiced without these specific details. In other cases, well-known methods, procedures, and components are not described in detail to avoid obscuring the present invention.

[0039] Unless otherwise defined herein, scientific and technical terms used in connection with the present application shall have the meanings commonly understood by those of ordinary skill in the art. In addition, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular.

[0040] A skilled person will understand that the term "comprising" encompasses the inclusion of stated elements but does not exclude other elements that may be optional.

[0041] A skilled person will understand that when a range of values ​​is expressed, another embodiment includes one limit and / or another limit of the range. In addition, all ranges are inclusive and combinable.

[0042] Treatment

[0043] The inventors surprisingly found that, contrary to the known assumption that several amyloid aggregation inhibitor molecules are needed to inhibit one amyloid β molecule, in the case of compound (1), the stoichiometry is exactly the opposite. For the old compound GAL-101 (MRZ-99030), which is a structurally related and previously best-in-class amyloid β aggregation inhibitor, it was found that at least 10 GAL-101 molecules were required to inhibit one amyloid β molecule (stoichiometric ratio of 10:1). For other similar compounds such as ALZ-801 and PRI-002, hundreds or thousands of molecules of detoxifying compounds are required to inhibit the toxicity caused by one amyloid β molecule (stoichiometric ratios ranging from about 100:1 to 5000:1). The inventors of the present disclosure found that one molecule of compound (1) can inhibit ten (!) amyloid β molecules, or even more (stoichiometric ratio of 1:10). The clinical significance of this surprising and surprising phenomenon is that the dose of compound (1) can be reduced by 90%, even 99% - to achieve the desired clinical effect, and as summarized in Table 1, the detoxification effect of compound (1) in the case of 10-fold undersupply (stoichiometric ratio 1:10) is even more than twice as strong as the application of stoichiometric overdose.

[0044] In some embodiments, disclosed herein is a method of treating or preventing a protein misfolding and deposition disease in a subject in need thereof, the method comprising administering to the subject a low dose of compound (1)

[0045]

[0046] Compound (1),

[0047] Thereby treating or preventing a protein misfolding and deposition disease in said subject.

[0048] In one embodiment, the term "treatment" refers to any process, action, application, therapy, etc., in which a subject, including humans, receives medical assistance with the goal of improving the subject's condition, either directly or indirectly.

[0049] In another embodiment, the term "treating" refers to reducing the incidence or alleviating symptoms, eliminating recurrence, preventing recurrence, preventing incidence, ameliorating symptoms, improving prognosis, or a combination thereof in other embodiments.

[0050] In another embodiment, "treating" includes ameliorating an existing condition. A skilled artisan will appreciate that treatment does not necessarily result in the complete disappearance or elimination of symptoms. Treatment also includes a palliative effect: that is, reducing the likelihood of a subsequent medical condition. The term encompasses a alleviation of a condition that leads to a more serious condition.

[0051] In one embodiment, "prevention" may encompass, among other things, delaying the onset of symptoms, preventing disease recurrence, reducing the number or frequency of recurrent episodes, increasing the latency period between symptom onsets, or a combination thereof.

[0052] The skilled artisan will appreciate that "protein misfolding and deposition diseases" occur when one of a growing list of proteins (amyloid-forming proteins in their native conformation, also known as amyloidogenic precursors) acquires an alternative folding state (a misfolded state), begins to aggregate and form oligomers, then fibrils and finally fibrous structures (called amyloid fibrils) that are ultimately deposited within tissues. Protein misfolding is believed to be the first step in the pathology of Alzheimer's disease, Creutzfeldt-Jakob disease and other degenerative and neurodegenerative disorders.

[0053] In some embodiments, disclosed herein is a method of inhibiting Aβ toxicity in a subject, the method comprising administering a low dose of compound (1)

[0054]

[0055] Compound (1),

[0056] Thereby inhibiting Aβ toxicity in the subject.

[0057] In one embodiment, low-dose administration results in an efficient supply-deficient ratio between compound (1) and amyloid beta in the tissue, resulting in a concentration of compound (1) at the site of action (typically in the brain or retina) of less than 50 nM. In another embodiment, low-dose administration results in a concentration of compound (1) at the site of action of less than 40 nM. In another embodiment, low-dose administration results in a concentration of compound (1) at the site of action of less than 30 nM. In another embodiment, low-dose administration results in a concentration of compound (1) at the site of action of less than 20 nM. In another embodiment, low-dose administration results in a concentration of compound (1) at the site of action of less than 10 nM. In another embodiment, low-dose administration results in a concentration of compound (1) at the site of action of less than 5 nM. In another embodiment, low-dose administration results in a concentration of compound (1) at the site of action of less than 5 nM. In another embodiment, low-dose administration results in a concentration of compound (1) at the site of action of less than 3 nM. In another embodiment, low-dose administration results in a concentration of compound (1) at the site of action of less than 1 nM.

[0058] In one embodiment, low dose administration results in a concentration of 50 nM of compound (1) at the site of action. In another embodiment, low dose administration results in a concentration of 40 nM of compound (1) at the site of action. In another embodiment, low dose administration results in a concentration of 30 nM of compound (1) at the site of action. In another embodiment, low dose administration results in a concentration of 20 nM of compound (1) at the site of action. In another embodiment, low dose administration results in a concentration of 10 nM of compound (1) at the site of action. In another embodiment, low dose administration results in a concentration of 5 nM of compound (1) at the site of action. In another embodiment, low dose administration results in a concentration of 5 nM of compound (1) at the site of action. In another embodiment, low dose administration results in a concentration of 3 nM of compound (1) at the site of action. In another embodiment, low dose administration results in a concentration of 1 nM of compound (1) at the site of action.

[0059] The term "site of action" is understood by those skilled in the art to be the site in the human body where disease pathology occurs. In one embodiment, where the site of action of the disease is the brain, for example, during a Phase 1 dose-finding study, the active concentration of compound (1) in the brain can be determined in cerebrospinal fluid (CSF) obtained from humans via lumbar puncture. The targeted oral dose of compound (1) is a dose corresponding to a 10 nM CSF concentration.

[0060] In some embodiments, the administration is oral administration.

[0061] In some embodiments, wherein the administration is oral administration, the low dose contains 10 mg or less of compound (1). In one embodiment, wherein the administration is oral administration, the low dose contains 10 mg of compound (1). In another embodiment, wherein the administration is oral administration, the low dose contains 9.0 mg of compound (1). In another embodiment, wherein the administration is oral administration, the low dose contains 8.0 mg of compound (1). In another embodiment, wherein the administration is oral administration, the low dose contains 7.0 mg of compound (1). In another embodiment, wherein the administration is oral administration, the low dose contains 6.0 mg of compound (1). In another embodiment, wherein the administration is oral administration, the low dose contains 5.0 mg of compound (1). In another embodiment, wherein the administration is oral administration, the low dose contains 4.0 mg of compound (1). In another embodiment, wherein the administration is oral administration, the low dose contains 3.0 mg of compound (1).

[0062] In some embodiments, the administration is intravascular (IV) administration.

[0063] In some embodiments, wherein the administration is IV administration, the low dose comprises 3 mg or less of compound (1). In one embodiment, wherein the administration is IV administration, the low dose comprises 3 mg of compound (1). In another embodiment, wherein the administration is IV administration, the low dose comprises 2.5 mg of compound (1). In another embodiment, wherein the administration is IV administration, the low dose comprises 2.0 mg of compound (1). In another embodiment, wherein the administration is IV administration, the low dose comprises 1.5 mg of compound (1). In another embodiment, wherein the administration is IV administration, the low dose comprises 1.0 mg of compound (1).

[0064] In some embodiments, disclosed herein is a method for reversing or preventing the toxic effects of misfolded and aggregated protein amyloid β, i.e., toxic Aβ oligomers, by using a relatively insufficient supply (reverse stoichiometric ratio) of compound (1) to achieve a stronger detoxification effect.

[0065] Compound (1)

[0066] Compound (1), referred to as GAL-201 (also previously known as MRZ-14042), is a proprietary small molecule having the following IUPAC nomenclature: (2R)-2-amino-N-(1-carbamoyl-1-methylethyl)-3-(1H-indol-3-yl)propanamide. The molecular formula of the free base is C 15 H 20N4O2, and the molecular weight is 288.34 g / Mol.

[0067] Compound (1) is represented by the following formula

[0068]

[0069] Compound (1) is structurally related to the best compound of the same type, GAL-101. GAL-101 is 2-[2-amino-3-(1H-indol-3-yl)-propionylamino]-2-methylpropanoic acid, C 15 H 19 N3O3, 289.33g / Mol. It has an effect on amyloid βAβ 1-42 The binding affinity was 28.9±2.9 nM.

[0070] Protein misfolding and deposition diseases

[0071] In some embodiments, the present disclosure relates to a method of treating or preventing a protein misfolding and deposition disease in a subject in need thereof. In some embodiments, the protein misfolding and deposition disease is associated with misfolding and deposition of amyloid beta. In some embodiments, amyloid beta includes Aβ 1-42 , Aβ 1-40 , Aβ p3-42 , its derivatives or any combination thereof.

[0072] In some embodiments, the protein misfolding and deposition disease is selected from a disease associated with amyloid beta pathology, i.e., Alzheimer's disease (AD), early-onset Alzheimer's disease, late-onset Alzheimer's disease, presymptomatic Alzheimer's disease, type II diabetes, serum amyloid A disease (SAA amyloidosis), hereditary Iceland syndrome, multiple myeloma, medullary carcinoma, aortic amyloidosis, cardiac amyloidosis, insulin injection amyloidosis, prion systemic amyloidosis, chronic inflammatory amyloidosis, senile systemic amyloidosis, pituitary amyloidosis, hereditary renal amyloidosis and familial non-neurological amyloidosis.

[0073] In one embodiment, protein misfolding and deposition diseases include Alzheimer's disease (AD). In another embodiment, protein misfolding and deposition diseases include early-onset Alzheimer's disease. In another embodiment, protein misfolding and deposition diseases include late-onset Alzheimer's disease. In another embodiment, protein misfolding and deposition diseases include presymptomatic Alzheimer's disease. In another embodiment, protein misfolding and deposition diseases include type II diabetes. In another embodiment, protein misfolding and deposition diseases include serum amyloid A disease (SAA amyloidosis). In another embodiment, protein misfolding and deposition diseases include hereditary Iceland syndrome. In another embodiment, protein misfolding and deposition diseases include insulin injection amyloidosis.

[0074] In another embodiment, the protein misfolding and deposition disease comprises multiple myeloma. In another embodiment, the protein misfolding and deposition disease comprises medullary carcinoma. In another embodiment, the protein misfolding and deposition disease comprises aortic amyloidosis. In another embodiment, the protein misfolding and deposition disease comprises cardiac amyloidosis. In another embodiment, the protein misfolding and deposition disease comprises insulin injection amyloidosis. In another embodiment, the protein misfolding and deposition disease comprises prion systemic amyloidosis. In another embodiment, the protein misfolding and deposition disease comprises chronic inflammatory amyloidosis. In another embodiment, the protein misfolding and deposition disease comprises senile systemic amyloidosis. In another embodiment, the protein misfolding and deposition disease comprises pituitary amyloidosis.

[0075] In another embodiment, the protein misfolding and deposition disease comprises hereditary renal amyloidosis.

[0076] In another embodiment, the protein misfolding and deposition disease comprises familial non-neuronal amyloidosis.

[0077] In another embodiment, the protein misfolding and deposition disease comprises Parkinson's disease, Huntington's disease, Creutzfeldt-Jakob disease, prion disease, or mad cow disease. In another embodiment, the protein misfolding and deposition disease comprises Parkinson's disease. In another embodiment, the protein misfolding and deposition disease comprises Huntington's disease. In another embodiment, the protein misfolding and deposition disease comprises Creutzfeldt-Jakob disease. In another embodiment, the protein misfolding and deposition disease comprises prion disease. In another embodiment, the protein misfolding and deposition disease comprises mad cow disease.

[0078] In another embodiment, the protein misfolding and deposition disease comprises amyloid-related eye diseases and disorders. In another embodiment, the protein misfolding and deposition disease comprises a prion disease.

[0079] In another embodiment, the protein misfolding and deposition disease comprises an amyloid-related disease.

[0080] The skilled person understands an "amyloid-related disease" as a pathology which is characterized by the accumulation and deposition of amyloid in certain tissues of the body, accompanied by functional defects of the corresponding organs.

[0081] In one embodiment, wherein the amyloid-related disease comprises Alzheimer's disease (AD), early-onset Alzheimer's disease, late-onset Alzheimer's disease, presymptomatic Alzheimer's disease, or any combination thereof, the treatment or prevention comprises improving cognitive deficits, improving memory loss, reducing abnormal behavior, reducing hallucinations, reducing spatial orientation loss, reducing apraxia, reducing aggression, improving the ability to perform daily living activities, or other symptoms of dementia, or any combination thereof, in the subject. In another embodiment, the treatment or prevention comprises improving cognitive deficits. In another embodiment, the treatment or prevention comprises improving memory loss. In another embodiment, the treatment or prevention comprises reducing abnormal behavior. In another embodiment, the treatment or prevention comprises reducing hallucinations. In another embodiment, the treatment or prevention comprises reducing spatial orientation loss. In another embodiment, the treatment or prevention comprises reducing apraxia. In another embodiment, the treatment or prevention comprises reducing aggression. In another embodiment, the treatment or prevention comprises improving the ability to perform daily living activities. In another embodiment, the treatment or prevention comprises improving other symptoms of dementia.

[0082] In another embodiment, the amyloid-related disease comprises glaucoma or age-related macular degeneration (AMD). In another embodiment, the amyloid-related disease comprises glaucoma. In another embodiment, the amyloid-related disease comprises age-related macular degeneration (AMD). In another embodiment, the amyloid-related disease comprises geographic atrophy (GA) secondary to AMD.

[0083] Skilled artisans understand glaucoma to be a group of eye diseases that cause damage to the optic nerve (or retina) and lead to vision loss. Risk factors for glaucoma include increasing age, high pressure in the eye, a family history of glaucoma, and the use of steroid medications.

[0084] In one embodiment, the glaucoma is selected from primary angle-closure glaucoma, open-angle glaucoma (primary and secondary), wide-angle glaucoma, steroid-induced glaucoma, traumatic glaucoma, secondary angle-closure glaucoma, and neovascular glaucoma.

[0085] In another embodiment, glaucoma comprises primary angle-closure glaucoma. In another embodiment, glaucoma comprises secondary open-angle glaucoma. In another embodiment, glaucoma comprises primary open-angle glaucoma. In another embodiment, glaucoma comprises wide-angle glaucoma. In another embodiment, glaucoma comprises steroid-induced glaucoma. In another embodiment, glaucoma comprises traumatic glaucoma. In another embodiment, glaucoma comprises secondary angle-closure glaucoma. In another embodiment, glaucoma comprises neovascular glaucoma.

[0086] The skilled artisan will appreciate that age-related macular degeneration (AMD) is a medical condition that can cause blurred or no vision in the center of the visual field. There are usually no symptoms in the early stages. However, over time, some people experience a gradual deterioration of vision that may affect one or both eyes. While it does not cause complete blindness, loss of central vision can make it difficult for a person to recognize faces, drive, read, or perform other daily activities. Visual hallucinations may also occur, but these do not represent a psychiatric disorder.

[0087] In one embodiment, AMD is selected from dry form and wet form. In another embodiment, AMD is dry form. In another embodiment, AMD is wet form. It will be appreciated by those skilled in the art that the difference between these two forms is the change of macula lutea. People suffering from dry form AMD may have drusen in their macula lutea, which usually grows over time and is relevant to the damage and loss of vision of photosensitive cells (photoreceptors and supporting pigment epithelial cells). When people suffer from wet form AMD, blood vessels grow under their macula lutea, which causes blood and fluid to leak into their retina.

[0088] In one embodiment, the amyloid-related disease is diabetic retinopathy. In another embodiment, the diabetic retinopathy comprises non-proliferative diabetic retinopathy, proliferative diabetic retinopathy, or a combination thereof.

[0089] In one embodiment, the protein misfolding and deposition disease is diabetic retinopathy, which can range from non-proliferative diabetic retinopathy and its stages to proliferative diabetic retinopathy. As the disease progresses, associated diabetic macular edema may also become apparent. This retinal disease may or may not be associated with the deposition of amyloid beta.

[0090] Dosage and Administration

[0091] In some embodiments, the administration is systemic administration.

[0092] In one embodiment, the systemic administration is by oral administration, rectal administration, transmucosal administration, nasal administration, intramuscular administration, subcutaneous administration, intrathecal administration, direct intracerebroventricular administration, intravenous administration, intraperitoneal administration, or intranasal instillation.

[0093] In another embodiment, systemic administration is by oral administration. In another embodiment, systemic administration is by rectal administration. In another embodiment, systemic administration is by transmucosal administration. In another embodiment, systemic administration is by nasal administration. In another embodiment, systemic administration is by intramuscular administration. In another embodiment, systemic administration is by subcutaneous administration. In another embodiment, systemic administration is by intrathecal administration. In another embodiment, systemic administration is by direct intracerebroventricular administration. In another embodiment, systemic administration is by intravenous administration. In another embodiment, systemic administration is by intraperitoneal administration. In another embodiment, systemic administration is by intranasal instillation.

[0094] In one embodiment, the oral administration comprises a dose of compound (1) in the range of 0.03-0.3 mg / kg. In another embodiment, the oral administration comprises a dose of compound (1) at 0.03 mg / kg. In another embodiment, the oral administration comprises a dose of compound (1) at 0.04 mg / kg. In another embodiment, the oral administration comprises a dose of compound (1) at 0.05 mg / kg. In another embodiment, the oral administration comprises a dose of compound (1) at 0.06 mg / kg. In another embodiment, the oral administration comprises a dose of compound (1) at 0.07 mg / kg. In another embodiment, the oral administration comprises a dose of compound (1) at 0.08 mg / kg. In another embodiment, the oral administration comprises a dose of compound (1) at 0.09 mg / kg. In another embodiment, the oral administration comprises a dose of compound (1) at 0.1 mg / kg. In another embodiment, the oral administration comprises a dose of compound (1) at 0.12 mg / kg. In another embodiment, the oral administration comprises a dose of compound (1) at 0.13 mg / kg. In another embodiment, the oral administration comprises a dose of compound (1) at 0.14 mg / kg. In another embodiment, the oral administration comprises a dose of 0.15 mg / kg of compound (1). In another embodiment, the oral administration comprises a dose of 0.16 mg / kg of compound (1). In another embodiment, the oral administration comprises a dose of 0.17 mg / kg of compound (1). In another embodiment, the oral administration comprises a dose of 0.18 mg / kg of compound (1). In another embodiment, the oral administration comprises a dose of 0.19 mg / kg of compound (1). In another embodiment, the oral administration comprises a dose of 0.2 mg / kg of compound (1). In another embodiment, the oral administration comprises a dose of 0.21 mg / kg of compound (1). In another embodiment, the oral administration comprises a dose of 0.22 mg / kg of compound (1). In another embodiment, the oral administration comprises a dose of 0.23 mg / kg of compound (1). In another embodiment, the oral administration comprises a dose of 0.24 mg / kg of compound (1). In another embodiment, the oral administration comprises a dose of 0.25 mg / kg of compound (1). In another embodiment, the oral administration comprises a dose of 0.26 mg / kg of compound (1). In another embodiment, the oral administration comprises a dose of 0.27 mg / kg of compound (1). In another embodiment, the oral administration comprises a dose of 0.28 mg / kg of compound (1). In another embodiment, the oral administration comprises a dose of 0.29 mg / kg of compound (1). In another embodiment, the oral administration comprises a dose of 0.3 mg / kg of compound (1).

[0095] In one embodiment, the intravenous administration comprises a dose of compound (1) in the range of 0.01-0.1 mg / kg. In one embodiment, the intravenous administration comprises a dose of compound (1) of 0.01 mg / kg. In another embodiment, the intravenous administration comprises a dose of compound (1) of 0.02 mg / kg. In another embodiment, the intravenous administration comprises a dose of compound (1) of 0.03 mg / kg. In another embodiment, the intravenous administration comprises a dose of compound (1) of 0.04 mg / kg. In another embodiment, the intravenous administration comprises a dose of compound (1) of 0.05 mg / kg. In another embodiment, the intravenous administration comprises a dose of compound (1) of 0.06 mg / kg. In another embodiment, the intravenous administration comprises a dose of compound (1) of 0.07 mg / kg. In another embodiment, the intravenous administration comprises a dose of compound (1) of 0.08 mg / kg. In another embodiment, the intravenous administration comprises a dose of compound (1) of 0.09 mg / kg. In another embodiment, the intravenous administration comprises a dose of compound (1) of 0.1 mg / kg.

[0096] In some embodiments, the administration is ocular administration.

[0097] In one embodiment, ocular administration is by eye drops, eye creams, eye ointments, eye sprays, intraocular reservoir formulations, by injection including intraocular injection, intraocular or periocular injection including intraocular fluids, reservoir formulations, solid and semisolid intraocular vehicles and matrix, or ophthalmic devices, such as contact lenses and films. In another embodiment, ocular administration is by eye drops. In another embodiment, ocular administration is by eye creams. In another embodiment, ocular administration is by eye ointments. In another embodiment, ocular administration is by eye sprays. In another embodiment, ocular administration is by intraocular reservoir formulations. In another embodiment, ocular administration is by injection including intraocular injection, intraocular or periocular injection including intraocular fluids, reservoir formulations, solid and semisolid intraocular vehicles and matrix. In another embodiment, ocular administration is through ocular devices, such as contact lenses and films.

[0098] In one embodiment, ocular administration comprises a single dose of less than 1 mg of compound (1) per eye. In another embodiment, ocular administration comprises a single dose of less than 0.8 mg of compound (1) per eye. In another embodiment, ocular administration comprises a single dose of less than 0.6 mg of compound (1). In another embodiment, ocular administration comprises a single dose of less than 0.4 mg of compound (1). In another embodiment, ocular administration comprises a single dose of less than 0.2 mg of compound (1). The above single doses are usually repeated one to three times a day during long-term treatment, up to lifelong treatment.

[0099] In one embodiment, the method comprises administering to the subject an initial loading dose of compound (1), and further administering to the subject a plurality of subsequent maintenance doses of compound (1). In one embodiment, the subsequent maintenance doses are lower doses of compound (1).

[0100] In one embodiment, the initial loading dose is administered 1-3 times per day for 1 day to 1 month.

[0101] In another embodiment, the initial loading dose is administered once a day for 1 day to 1 month. In another embodiment, the initial loading dose is administered twice a day for 1 day to 1 month. In another embodiment, the initial loading dose is administered 3 times a day for 1 day to 1 month.

[0102] In another embodiment, the initial loading dose is administered 1-3 times per day for 1 day. In another embodiment, the initial loading dose is administered 1-3 times per day for 5 days. In another embodiment, the initial loading dose is administered 1-3 times per day for 10 days. In another embodiment, the initial loading dose is administered 1-3 times per day for 15 days. In another embodiment, the initial loading dose is administered 1-3 times per day for 20 days. In another embodiment, the initial loading dose is administered 1-3 times per day for 25 days. In another embodiment, the initial loading dose is administered 1-3 times per day for 30 days.

[0103] In one embodiment, the maintenance dose is administered once a week, once every two weeks, or once every four weeks. In another embodiment, the maintenance dose is administered once a week. In another embodiment, the maintenance dose is administered once every two weeks. In another embodiment, the maintenance dose is administered once every four weeks.

[0104] In one embodiment, the maintenance dose is 10%-75% of the initial loading dose. In another embodiment, the maintenance dose is 10% of the initial loading dose. In another embodiment, the maintenance dose is 20% of the initial loading dose. In another embodiment, the maintenance dose is 30% of the initial loading dose. In another embodiment, the maintenance dose is 40% of the initial loading dose. In another embodiment, the maintenance dose is 50% of the initial loading dose. In another embodiment, the maintenance dose is 60% of the initial loading dose. In another embodiment, the maintenance dose is 70% of the initial loading dose. In another embodiment, the maintenance dose is 75% of the initial loading dose.

[0105] In one embodiment, the compound is administered to the subject daily for a first period of at least one day, followed by a second period of at least one week in which the compound is not administered, followed by a repetition of the at least one day first period in which the subject is administered the compound daily.

[0106] Pharmaceutical composition

[0107] The compositions described herein can be incorporated into pharmaceutical compositions suitable for administration. In one embodiment, a "pharmaceutical composition" or "pharmaceutical formulation" refers to a preparation of one or more active ingredients described herein with other chemical components (such as physiologically suitable carriers and excipients). The purpose of a pharmaceutical composition or "pharmaceutical formulation" is to facilitate the administration of a compound to a subject. In certain embodiments, a "pharmaceutical composition" or "pharmaceutical formulation" provides a pharmaceutical dosage form of a drug.

[0108] In some embodiments, "active ingredient" refers to the molecule responsible for the biological effect. In some embodiments, the active ingredient comprises compound (1).

[0109] In some embodiments, the pharmaceutical composition comprises an excipient. A skilled artisan will appreciate that in some embodiments, the excipient is selected based on the mode of delivery of the pharmaceutical composition.

[0110] In one embodiment, compound (1) is the active ingredient of a pharmaceutical composition, which further comprises a physiologically acceptable carrier.

[0111] Pharmaceutical compositions typically include a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc. that are compatible with drug administration. Suitable carriers are described in the latest edition of Remington's Pharmaceutical Sciences, a standard reference text in the field, which is incorporated herein by reference.

[0112] Some examples of substances that can be used as pharmaceutically acceptable carriers or components thereof are sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and methylcellulose; powdered tragacanth; malt; gelatin; talc; solid lubricants such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and cocoa butter; polyols such as propylene glycol, glycerol, sorbitol, mannitol and polyethylene glycol (PEG); alginic acid; emulsifiers such as Tween TM Brand emulsifiers; wetting agents, such as sodium lauryl sulfate; colorants; flavoring agents; tableting agents, stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline; and phosphate buffered saline. The choice of a pharmaceutically acceptable carrier to be used in conjunction with the compound is essentially determined by the manner in which the compound is to be administered. If the test compound is to be injected, in one embodiment, the pharmaceutically acceptable carrier is sterile physiological saline containing a blood compatible suspending agent, the pH of which has been adjusted to about 7.4.

[0113] In some embodiments, the pharmaceutical composition further comprises a binder (e.g., gum arabic, corn starch, gelatin, carbomer, ethyl cellulose, guar gum, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, povidone), a disintegrant (e.g., corn starch, potato starch, alginic acid, silicon dioxide, cross-linked carboxymethyl cellulose sodium, cross-linked povidone, guar gum, sodium starch glycolate), a buffer of various pH and ionic strength (e.g., Tris-HCl., acetate, phosphate), an additive to prevent surface absorption (e.g., albumin or gelatin), a detergent (e.g., Tween 20, Tween 80, Pluronic F68, bile acid salts), protease inhibitors, surfactants (e.g., sodium lauryl sulfate), penetration enhancers, solubilizers (e.g., glycerol, polyethylene glycerol), antioxidants (e.g., ascorbic acid, sodium metabisulfite, butylated hydroxyanisole), stabilizers (e.g., hydroxypropyl cellulose, hydroxypropyl methylcellulose), viscosity increasing agents (e.g., carbomer, colloidal silicon dioxide, ethyl cellulose, guar gum), sweeteners (e.g., aspartame, citric acid), preservatives (e.g., thimerosal, benzyl alcohol, parabens), lubricants (e.g., stearic acid, magnesium stearate, polyethylene glycol (PEG), sodium lauryl sulfate), glidants (e.g., colloidal silicon dioxide), plasticizers (e.g., diethyl phthalate, triethyl citrate), emulsifiers (e.g., carbomer, hydroxypropyl cellulose, sodium lauryl sulfate), polymer coatings (e.g., poloxamer or polyoxamine), coatings and film formers (e.g., ethyl cellulose, acrylates, polymethacrylates), adjuvants, or any combination thereof.

[0114] In one embodiment, the pharmaceutical composition is suitable for oral administration, rectal administration, transmucosal administration, nasal administration, intramuscular administration, subcutaneous administration, intrathecal administration, direct intracerebroventricular administration, intravenous administration, intraperitoneal administration, intranasal instillation, or ophthalmic administration.

[0115] In one embodiment, the pharmaceutical composition is suitable for oral administration. In another embodiment, the pharmaceutical composition is suitable for rectal administration. In another embodiment, the pharmaceutical composition is suitable for transmucosal administration. In another embodiment, the pharmaceutical composition is suitable for nasal administration. In another embodiment, the pharmaceutical composition is suitable for intramuscular administration. In another embodiment, the pharmaceutical composition is suitable for subcutaneous administration. In another embodiment, the pharmaceutical composition is suitable for intrathecal administration. In another embodiment, the pharmaceutical composition is suitable for direct intracerebroventricular administration. In another embodiment, the pharmaceutical composition is suitable for intravenous administration. In another embodiment, the pharmaceutical composition is suitable for intraperitoneal administration. In another embodiment, the pharmaceutical composition is suitable for intranasal instillation. In another embodiment, the pharmaceutical composition is suitable for ocular administration.

[0116] In some embodiments, compositions comprising a formulation disclosed herein formulated in a compatible pharmaceutical carrier are also prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

[0117] The skilled artisan will appreciate that foam compositions are typically formulated in a single-phase or multi-phase liquid form and contained in a suitable container, optionally together with a propellant, which aids in expelling the composition from the container, thereby converting it into a foam when applied. Other foam forming techniques include, for example, "bag-in-can" formulation techniques. Compositions so formulated typically contain low boiling hydrocarbons, such as isopropyl alcohol. Application and agitation of such compositions at body temperature causes the isopropyl alcohol to evaporate and produce foam in a manner similar to pressurized aerosol foaming systems. Foams can be water-based or hydroalcoholic, but are typically formulated to have a high alcohol content, which evaporates rapidly when applied to the user's skin, thereby driving the active ingredient through the upper layers of the skin to the treatment site.

[0118] In one embodiment, the formulation provided herein also includes preservatives such as benzalkonium chloride and thimerosal, etc.; chelating agents such as sodium edetate, etc.; buffers such as phosphates, citrates and acetates; tonicity agents such as sodium chloride, potassium chloride, glycerol, mannitol, etc.; antioxidants such as ascorbic acid, acetylcysteine, sodium pyrosulfite, etc.; fragrances; viscosity modifiers such as polymers including cellulose and its derivatives; and polyvinyl alcohol and acids and bases to adjust the pH of these aqueous compositions as needed. The composition also includes a local anesthetic or other active ingredients. The composition can be used as a spray, mist, drops, etc.

[0119] Example

[0120] Example 1: Compound (1) targets amyloid βAβ 1-42 affinity.

[0121] The purpose of this study was to evaluate the effect of compound (1) on misfolded Aβ 1-42 The target affinity of the monomer. The affinity is determined using in vitro surface plasmon resonance (SPR) technology. SPR experiments allow the study of the binding of compounds to lower concentrations of Aβ 1-42 The Biacore X100 SPR instrument is equipped with two flow cells on a sensor chip for real-time binding studies.

[0122] Aβ 1-42 (American Peptide Company, Sunnyvale, CA, USA) was dissolved in hexafluoroisopropanol (HFIP) to 1 mg / ml. The tubes were incubated at room temperature with shaking for 1.5 hours, 100 μg aliquots were prepared in low-binding Eppendorf tubes, and frozen at -80°C for 30-60 min. After lyophilization overnight, the aliquots were stored at -20°C until use. To prepare monomers, an HFIP-treated Aβ aliquot was thawed and freshly dissolved in DMSO (anhydrous), the 5 mM stock solution was centrifuged (5 min 13000 g), and the supernatant was diluted to 100 μM in 10 mM sodium acetate at pH 4.0 immediately before fixation.

[0123] Aβ monomers were covalently coupled to the flow cell of a CM7 sensor chip (carboxymethylated dextran matrix attached to a gold surface). 1-42 The density is about 1pg / mm 2 To fix human Aβ 1-42 Monomeric, HFIP-treated peptides were dissolved in DMSO to 5 mM, diluted to 100 μM in 10 mM sodium acetate, pH 4.0, and immediately coupled to one flow cell surface of the sensor chip. A second flow cell was used as a reference and treated with ethanolamine instead of Aβ. To determine affinity, compounds (1) were tested at concentrations ranging from 0.3 nM up to 1000 nM using HBS-EP, 0.1% DMSO as running buffer at 25°C.

[0124] The resonance units (RU) induced by the compound injected into the ethanolamine control flow cell were set as the reference response and subtracted from the RU induced by the same compound injected into the Aβ saturated flow cell. Each RU obtained at the binding steady state (plateau of the binding curve) was plotted against each concentration of the compound. Biacore X100 software Ver 1.1 was used to record the binding curves and analyze them (plot each RU at steady state versus analyte concentration, fit the plot, determine K D The dissociation equilibrium constant K of the analyte relative to the immobilized Aβ D Determined by the steady-state level, the maximum RUR is estimated max , and K D Calculated as half of the R max concentration of the compound.

[0125] Compound (1) K at Aβ monomer D The results are presented in Figure 1 Medium. The affinity for the target is 4 times stronger than that of the previous best compound in its class, GAL-101.

[0126] Example 2: Pharmacokinetics of compound (1) in rat brain.

[0127] The purpose of this experiment is to evaluate the pharmacokinetics of compound (1) in rat brain. Compound (1) was administered subcutaneously to anesthetized or awake and freely moving animals. A microdialysis probe was placed in the prefrontal cortex of these adult male Sprague Dawley rats to measure the concentration of compound (1) in brain interstitial fluid (ISF) using an HPLC system with an MS / MS detector.

[0128] The results are presented in Figure 2 middle.

[0129] Example 3: Dose-dependent effects of compound (1) at very low concentrations on LTP in anesthetized rats.

[0130] The purpose of this study was to evaluate the dose-dependent effect of compound (1) on LTP in anesthetized rats at very low concentrations. Compound (1) was administered sc at doses of 0.08 mg / kg, 0.4 mg / kg, and 2.0 mg / kg. Amyloid βAβ was injected into the cytoplasm at a concentration that strongly inhibited LTP. 1-42 Inject into the cerebral ventricle. Figure 3 As shown, surprisingly, a putative inactive dose of only 0.4 mg / kg (medium grey), resulting in a peak brain concentration of 10 nM, showed at least as potent efficacy on LTP.

[0131] Example 4: Dose-dependent effects of compound (1) on in vitro LTP assay.

[0132] For long-term potentiation (LTP) experiments, brain slices from 2-month-old mice were used. After decapitation under isoflurane anesthesia, transverse hippocampal slices (350 μm thick) were obtained. All slices were placed in a holding chamber for at least 60 min and then transferred to a superfusion chamber for extracellular recording. The flow rate of the solution through the chamber was 4 ml / min. The composition of the solution was 124 mM NaCl, 3 mM KCl, 26 mM NaHCO3, 2 mM CaCl2, 1 mM MgSO4, 10 mM D-glucose and 1.25 mM NaH2PO4, bubbled with a 95% O2 / 5% CO2 mixture, and the final pH was 7.3. All experiments were performed at room temperature. Aβ in DMSO was added to the superfusion chamber. 1-42 The stock solution was added to the bath solution so that the final concentration was 50 nM. Extracellular recordings of field excitatory postsynaptic potentials (fEPSPs) were obtained from the dendritic region of the CA1 region of the hippocampus using a glass micropipette filled with superperfusion solution. For all recordings, two stimulating electrodes were used to exploit the input specificity of LTP, allowing internal controls to be measured within the same slice. A stable baseline recording of at least 30 minutes was performed before applying tetanic stimulation. For LTP induction, high-frequency stimulation (HFS) conditioning pulses (100 Hz / 1 s; 4-5 V) were applied to the Schaffer collateral-commissural pathway via two independent inputs. Before inducing LTP, Aβ was applied. 1-42 90 min. HFS was delivered from one of the electrodes in the presence of compound (1) and the potentiation of the response was monitored for at least 60 min after tetanus. Aβ was then applied via the bath 1-42 90 min, then attempted to induce LTP in the second input after HFS was delivered via the second electrode. The amplified fEPSPs were filtered (3 kHz), digitized (15 kHz), measured and plotted ( Figure 4A fEPSP slope was measured between 20% and 80% of the peak amplitude. The fEPSP slope was normalized to the 20 min control period before tetanic stimulation.

[0133] In these LTP experiments, compound (1) was used at different concentrations. Surprisingly, when compound (1) was applied at the lowest concentration of only 10 nM, it best prevented the 50 nM Aβ 1-42 Detrimental effects on hippocampal LTP. Figure 4B Summarizes the causes of different stoichiometric Aβ 1-42Effect of different compound (1) concentrations at the ratio (10:1, 2:1, 1:5) of 1:1 / compound (1). LTP signals (fEPSP enhancement) calculated as the average of the last 50-60 min after HFS are presented in the form of bar graphs.

[0134] Compound (1) and amyloid protein βAβ 1-42 The reversal of the stoichiometric ratio between is presented in Table 1 below.

[0135]

[0136] Table 1 This table shows that compound (1) and amyloid protein βAβ 1-42 The reversal of the stoichiometric ratio between the two enhances the detoxification strength of compound (1). The data in this table are taken from Figure 4B The control experiment was defined as 100% normalized LTP response, and 50 nM Aβ 1-42 The toxic effect of oligomers is defined as 0% normalized LTP change. The use of the usual stoichiometric excess of compound (1), i.e. 500 nM or 100 nM, resulted in a weak detoxification effect of 40.2% and 34.7%, respectively. However, reversing the stoichiometric ratio and using compound (1) at a very low concentration (10 nM) (representing a 5-fold undersupply) significantly increased the detoxification intensity to 88.1%.

[0137] Control experiments were defined as 100% normalized LTP responses and at 50 nM Aβ 1-42 Under the toxic effect of oligomers, the normalized LTP change was defined as 0%. Using the usual stoichiometric excess of compound (1), i.e., 500nM or 100nM, a moderate detoxification effect of 40.2% and 34.7% was observed, respectively. However, reversing the stoichiometric ratio and using compound (1) at a concentration of only 10nM (representing a 5-fold stoichiometric undersupply) significantly increased the detoxification intensity, reaching an effect size of 88.1.

[0138] Combining calculations from in vitro and in vivo LTP experiments, it was demonstrated that a subcutaneous (sc) dose of 0.4 mg / kg of compound (1), which resulted in a concentration of 10 nM of compound (1) in the extracellular space of the brain, completely detoxified the currently toxic Aβ 1-42 , and leads to a fully restored LTP signal (see Figure 3 ).

[0139] Intracerebroventricular (icv) injection of 6 μL oligomeric Aβ 1-42 A significant lack of LTP was observed in animals treated with the solution (1.67 mM). Figure 3At the end of the recording period, i.e. 80–90 min after LTP induction, oligomeric Aβ was administrated icv 1-42 PS amplitudes measured after 14 h were 138.1 ± 4.1% of baseline (n = 18), compared with 179.2 ± 8.7% (n = 12) in animals injected with PBS. 1-42 In animals with icv injection of oligomeric Aβ 1-42 Previously, three different sc doses of compound (1) were administered, namely 0.08, 0.4 and 2 mg / kg. 0.4 mg / kg and 2 mg / kg of compound (1) resulted in post-TBS responses comparable to those observed in control animals (P<0.05, one-way ANOVA with post hoc Bonferroni test). At the end of the recording period, i.e. 80-90 min after LTP induction, sc administration of compound (1) and icv administration of oligomeric Aβ 1-42 PS amplitudes measured after 10 min were 170.0 ± 11.9% of baseline for the 2 mg / kg dose (n = 7), 174.7 ± 16.0% of baseline for the 0.4 mg / kg dose (n = 6), and 139.6 ± 13.1% of baseline for the 0.08 mg / kg dose (n = 4), compared to 179.2 ± 8.7% in the vehicle-treated control group (n = 12) and in the icv-injected oligomeric Aβ 1-42 These results confirm that compound (1) found in the above in vitro experiments inhibits Aβ 1-42 High potency of toxicity (presented in Figure 4B middle).

[0140] Although certain features of the invention have been illustrated and described herein, many modifications, substitutions, changes and equivalents will now occur to those skilled in the art. It should therefore be understood that the appended claims are intended to cover all such modifications and changes that fall within the true spirit of the invention.

Claims

1. A method for treating or preventing a protein misfolding and deposition disease in a subject in need thereof, the method comprising administering to the subject a low dose of a compound (1) Thereby treating or preventing a protein misfolding and deposition disease in said subject.

2. The method of claim 1, wherein the low dose administration results in a concentration of compound (1) at the site of action of less than 50 nM.

3. The method according to claims 1 and 2, wherein the low dose administration results in a concentration of 10 nM of compound (1) at the site of action.

4. The method according to claims 1 and 2, wherein the low dose administration results in a concentration of 3 nM of compound (1) at the site of action.

5. The method according to claims 1 and 2, wherein the low dose administration results in a concentration of 1 nM of compound (1) at the site of action.

6. The method of claim 1, wherein the protein misfolding and deposition disease is selected from the group consisting of Alzheimer's disease (AD), early-onset Alzheimer's disease, late-onset Alzheimer's disease, presymptomatic Alzheimer's disease, type II diabetes, serum amyloid A disease (SAA amyloidosis), hereditary Iceland syndrome, multiple myeloma, medullary carcinoma, aortic amyloidosis, cardiac amyloidosis, insulin injection amyloidosis, prion systemic amyloidosis, chronic inflammatory amyloidosis, senile systemic amyloidosis, pituitary amyloidosis, hereditary renal amyloidosis, and familial non-neurological amyloidosis.

7. The method of claim 1, wherein the protein misfolding and deposition disease is selected from the group consisting of Parkinson's disease, Huntington's disease, Creutzfeldt-Jakob disease, prion disease, and mad cow disease.

8. The method of claim 1, wherein the protein misfolding and deposition disease comprises an amyloid-related disease.

9. The method of claim 8, wherein the amyloid-related disease comprises Alzheimer's disease (AD), early-onset Alzheimer's disease, late-onset Alzheimer's disease, presymptomatic Alzheimer's disease, or any combination thereof.

10. The method of claim 9, wherein the treating or preventing comprises improving cognitive deficits, improving memory loss, reducing abnormal behavior, reducing hallucinations, reducing spatial orientation loss, reducing apraxia, reducing aggression, improving the ability to perform activities of daily living or other symptoms of dementia, or any combination thereof in the subject.

11. The method of claim 8, wherein the amyloid-related disease comprises diabetic retinopathy.

12. The method of claims 1-11, wherein the administration is systemic administration.

13. The method of claim 12, wherein the systemic administration is by oral administration, rectal administration, transmucosal administration, intranasal administration, intramuscular administration, subcutaneous administration, transdermal administration, intrathecal administration, direct intracerebroventricular administration, intravenous administration, intraperitoneal administration, or intranasal instillation.

14. The method of claim 13, wherein the oral administration comprises compound (1) at a dose in the range of 0.03-0.3 mg / kg.

15. The method of claim 13, wherein the intravenous administration comprises compound (1) at a dose in the range of 0.01-0.1 mg / kg.

16. The method of claim 8, wherein the amyloid-related disease is selected from glaucoma or age-related macular degeneration (AMD).

17. The method of claim 16, wherein the glaucoma is selected from primary angle-closure glaucoma, open-angle glaucoma (primary and secondary), wide-angle glaucoma, steroid-induced glaucoma, traumatic glaucoma, secondary angle-closure glaucoma, and neovascular glaucoma.

18. The method of claims 16-17, wherein the administration is ocular administration.

19. The method of claim 18, wherein the ocular administration is by eye drops, eye creams, eye ointments, eye sprays, intraocular depot formulations, by injection including intraocular injections, intraocular or periocular injections including intraocular fluids, depot formulations, solid and semisolid intraocular vehicles and matrices, or ophthalmic devices such as contact lenses and membranes.

20. The method of claim 19, comprising administering less than 1 mg of compound (1) as a single dose to the eye of a subject in need thereof.

21. The method of claims 1-20, wherein the method comprises administering to the subject an initial loading dose of compound (1), and further administering a plurality of subsequent maintenance doses of the compound (1).

22. The method of claim 21, wherein the initial loading dose is administered 1-3 times per day for 1 day to 1 month.

23. The method of claim 21, wherein the maintenance dose is administered once a week, once every two weeks, or once every four weeks.

24. The method of claims 21-23, wherein the maintenance dose is 10%-75% of the initial loading dose.

25. The method of claim 1, wherein the compound is administered to the subject daily for a first period of at least one day, followed by a second period of at least one week in which the compound is not administered, followed by a repetition of the at least one day first period in which the subject is administered the compound daily.

26. The method according to claim 1, wherein the compound (1) is an active ingredient of a pharmaceutical composition further comprising a physiologically acceptable carrier.

27. The method of claim 26, wherein the pharmaceutical composition is suitable for oral administration.

28. The method of claim 26, wherein the pharmaceutical composition is suitable for ocular administration.

29. A method for inhibiting Aβ toxicity in a subject, the method comprising administering a low dose of compound (1) Compound (1), Thereby inhibiting Aβ toxicity in the subject.

30. The method of claim 29, wherein the low dose administration results in a concentration at the site of action of less than 50 nM.

31. The method according to claims 29 and 30, wherein the low dose administration achieves a concentration of 10 nM at the site of action.

32. The method of claims 29 and 30, wherein the low dose administration achieves a concentration of 3 nM at the site of action.

33. The method according to claims 29 and 30, wherein the low dose administration achieves a concentration of 1 nM at the site of action.

34. A method for reversing or preventing the toxic effects of misfolded and aggregated protein amyloid beta, i.e., toxic A beta oligomers, by using a relatively insufficient supply (reverse stoichiometric ratio) of compound (1) to achieve a stronger detoxification effect.

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