Phosphated compounds for the detection of neurological diseases

Abstract

This document provides compounds, methods, and compositions for determining whether a patient has a neurological disease or condition, including detecting the presence or cumulative mass of a detectable target protein, such as amyloid-β protein or phosphorylated tau protein, in a patient's tissue or sample. The detection may include contacting the target protein with a compound described herein.

Description

[0001] Cross-references to related applications

[0002] Pursuant to section 119(e) of Chapter 35 of the United States Code, this application claims the benefit of U.S. Provisional Application No. 62 / 755,331, filed November 2, 2018, and U.S. Provisional Application No. 62 / 855,442, filed May 31, 2019, the entire contents of each of which are incorporated herein by reference. Background Technology

[0003] Neurological disorders are diseases of the brain, spinal cord, and peripheral nervous system. In terms of epidemiology and individual morbidity, the greatest social cost is borne by neurological diseases. Among these are Alzheimer's disease and Parkinson's disease. Other neurological disorders include age-related conditions (e.g., Parkinson's dementia, vascular dementia, amyotrophic lateral sclerosis), genetic syndromes (e.g., Down syndrome), injury-related conditions (e.g., traumatic brain injury, chronic traumatic encephalopathy), and conditions often considered purely psychological in nature, such as schizophrenia and depression.

[0004] Compounds used to detect these and other neurological disorders can be used for the detection, diagnosis, monitoring, and treatment of these diseases. Summary of the Invention

[0005] This disclosure provides compounds for the detection and treatment of neurological diseases or conditions. It has been found that certain diagnostic compounds used for the detection of neurological diseases have less than ideal physicochemical properties, such as low water solubility and high crystallinity. Compounds with improved physicochemical properties are disclosed herein, which can be used for the diagnosis and treatment of neurological diseases.

[0006] In some embodiments, a method is provided for determining whether a patient has a neurological disease or condition, the method comprising administering a compound described herein to the patient. In some embodiments, a method is provided for detecting a neurological disease or condition in a patient, the method comprising administering a compound described herein capable of binding to a detectable target protein, or a compound that, after separation, is capable of binding to a detectable target protein, to the patient; determining the presence or absence of binding, wherein the presence of binding indicates that the patient has a neurological disease or condition or is at risk of developing a neurological disease or condition. The compound may be a compound of formula I. In some embodiments, the compound is compound 7, compound 8, compound 9, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is compound 7, compound 8, compound 9, or compound 10. Therefore, methods and compositions are provided herein for determining whether a patient has a neurological disease or condition, the method comprising detecting the presence or cumulative mass (e.g., amyloid β protein or phosphorylated tau protein or their cumulative mass) of a detectable target protein in a patient's tissue or sample. The detection may include contacting the target protein with a compound described herein.

[0007] In some implementations, the patient has no symptoms of the disease or condition to be detected. In other implementations, the patient has one or more symptoms of the disease or condition to be detected.

[0008] In some embodiments, the method includes administering the compounds described herein to a patient. In some embodiments, the administration is intravenous or intraretinal. In some embodiments, the administration is a bolus injection. In some embodiments, the administration is a topical application.

[0009] In some embodiments, the neurological disease or condition is selected from age-related diseases or conditions, genetic diseases or conditions, injury-related diseases or conditions, and mental illnesses or conditions. In some embodiments, the age-related disease or condition is selected from Parkinson's disease, vascular dementia, and amyotrophic lateral sclerosis (ALS); the genetic disease or condition is Down syndrome; the injury-related disease or condition is selected from traumatic brain injury and chronic traumatic encephalopathy; and the mental illness or condition is selected from schizophrenia and depression. In some embodiments of the method, the neurological disease or condition is Alzheimer's disease or traumatic brain injury (TBI).

[0010] In some embodiments of the method, the neurological disease or condition is a prion disease. Therefore, a method is provided for detecting prion deposits in a patient. In some embodiments, the detection is performed in the retina. In some embodiments, a method is provided for detecting prion deposits in the retina of a patient, wherein the patient may or may not exhibit clinical manifestations of an associated disease or condition, such as Creutzfeldt-Jakob disease (e.g., behavioral changes, confusion, cognitive impairment, motor problems, visual impairment, kyphosis, ataxia, toe walking, etc.).

[0011] In some embodiments of the method, the neurological disease or condition is cerebral amyloid angiopathy (CAA). Cerebral amyloid angiopathy (CAA) is an age-related disease characterized by amyloid deposition within the walls of cerebral blood vessels. Therefore, in some embodiments, a method for detecting Aβ40 in a patient is provided. In some embodiments, this detection is performed in the retina.

[0012] In some implementations, the detection distinguishes between Aβ variant Aβ40 and variant Aβ42. Attached Figure Description

[0013] Figures 1A to 1C Compound 8 is shown. 1 H NMR spectrum.

[0014] Figures 2A to 2B Compound 9 is shown. 1 H NMR spectrum.

[0015] Figure 3 Images of real-time retinal imaging in mice before and after chronic sciatic nerve compression injury (CCI) using compound 10 are presented. Top row: Temporal retinal imaging of 3-month-old mice before CCI following intravenous administration of compound 10. The timelines marked at the top indicate in vivo retinal imaging time points before (t = 0) and after intravenous administration of compound 10. Bottom row: Temporal retinal imaging of the same mouse using compound 10 24 hours after CCI.

[0016] Figure 4 and Figure 5 It showed the symptoms before ( Figure 4 ) and the final stage ( Figure 5 Retinal fluorescence images of GPI-anchored transgenic mice. Images were acquired on the Phoenix MicronIV rodent retinal imaging system before (pre-injection, t = 0) and after (post-injection) systemic administration of compound 10. Brightened areas in the images represent regions where retinal virus deposition was observed.

[0017] Figure 6 A to Figure 6 C shows staining of amyloid deposits in the hippocampus of a patient with familial Alzheimer's disease. Figure 6 A shows staining with compound 7. Figure 6 B shows staining with 6E10 and anti-A antibody. Figure 6 C is a combined image of A and B, showing the co-staining of compound 7 and 6E10 (white arrows).

[0018] Figure 7 In vivo retinal images and ex vivo immunohistochemical (IHC) images from mice following intravenous administration of compound 10 are shown. Retinal images of APPSwDI (10 months old, female), PSAPP (10 months old, female), or wild-type (9 months old, female) mice at t = 0 (first column) or t = 5 minutes (second column) after intravenous injection of compound 10 are shown. Bright-field images (inset) or fluorescent retinal images were obtained using the Phoenix Micron IV rodent retinal imaging system. Last column: Ex vivo IHC staining of the retina with DAPI nuclear staining or 6E10 antibody. 。 Compound 7 was labeled from an in vivo injection of compound 10 (i.e., without further staining). Each channel is shown individually; all three channels are merged to show co-localization of compound 7 with 6E10. White arrows indicate areas of observed deposition.

[0019] Figure 8 The image shows the ex vivo immunohistochemistry (IHC) of brain slices from mice 15 minutes after intravenous administration of compound 10. The brain slices were stained with DAPI nuclear stain and 6E10. The labeling of compound 7 is from in vivo injection of compound 10. APPSwDI: White arrows indicate amyloid deposits along vessels labeled with compounds 7 and 6E10. PSAPP: Labeling of amyloid plaques with compounds 7 and 6E10 (white arrows).

[0020] Figure 9 This shows in vitro staining of human brain tissue from four different patients diagnosed with CAA and four healthy controls. 。 The staining methods are as follows: blue = DAPI nuclear staining, red = 6E10 staining, green / yellow = staining with compound 7. Scale bar = 50 µm. White arrows indicate areas where green / yellow staining was observed.

[0021] Figure 10 The in vivo detection of retinal α-syn deposits associated with Parkinson's disease is shown using the Line 61 model.

[0022] Figure 11The image shows retinal tissue extracted from mice stained with DAPI and SYN1 antibodies after necropsy.

[0023] Figure 12 This indicates that administration of compound 10 can reproducibly detect retinal α-syn deposits in the same animal.

[0024] Figure 13 This indicates that administration of compound 10 can detect retinal α-synuclein deposition at a low dose of 3 mg / kg.

[0025] Figure 14 An exemplary image from the right retina of a healthy person is shown.

[0026] Figure 15 An exemplary image from a healthy human left retina is shown.

[0027] Figure 16 Exemplary images of various regions of the right retina extracted from a 71-year-old male diagnosed with Parkinson's disease (PD) are shown, stained with compound 7 and SYN-1 Ab (α-synuclein).

[0028] Figure 17 Exemplary images of various regions of the left retina stained with compound 7 and SYN-1 Ab (α-synuclein) are shown from a 71-year-old man diagnosed with Parkinson's disease (PD).

[0029] Figure 18 This indicates that compound 7 co-localizes with SYN1 in all four quadrants of a pair of Parkinson's disease eyes. Detailed Implementation

[0030] definition

[0031] The following description sets forth exemplary embodiments of the present disclosure. However, it should be understood that this description is not intended to limit the scope of the present disclosure, but is provided as a description of exemplary embodiments.

[0032] The following words, phrases and symbols used in this specification are generally intended to have the meanings stated below, unless otherwise stated in the context in which they are used.

[0033] A dash (e.g., "-" or "-") indicates a bond, which may be a connection point for a substituent. For example, -C(O)NH2 is connected via carbon atoms. Chemical groups may be depicted with one or more dashes or without using one or more dashes without losing their ordinary meaning. A wavy line drawn using lines in the structure represents a connection point for a substituent or group.

[0034] prefix "C" u-v "" indicates that the following group has u to v carbon atoms. For example, "C 1-6 "Alkyl" indicates that the alkyl group has 1 to 6 carbon atoms.

[0035] References to the value or parameter “about” herein include (and describe) embodiments for that value or parameter itself. In some embodiments, the term “about” includes ±10% of the indicated amount. In other embodiments, the term “about” includes ±5% of the indicated amount. In some other embodiments, the term “about” includes ±1% of the indicated amount. Additionally, the term “about X” includes a description of “X”. Furthermore, unless explicitly stated otherwise in the context, the singular forms “a” and “the” include multiple references. Thus, for example, references to “compound” include a variety of such compounds, and references to “assay” include references to one or more assays and their equivalents known to those skilled in the art.

[0036] The term "alkyl" itself, or as part of another substituent, indicates a straight (i.e., unbranched) or branched chain, or a combination thereof, which can be fully saturated, monounsaturated, or polyunsaturated alkyl groups, and may include alkyl groups having a specified number of carbon atoms (i.e., C1-C1). 10 This refers to divalent and polyvalent groups (representing 1 to 10 carbons). Examples of saturated hydrocarbon groups include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, (cyclohexyl)methyl, for example, n-pentyl, n-hexyl, n-heptyl, n-octyl, etc.

[0037] "Alkenyl" refers to a group containing at least one carbon-carbon double bond and having 2 to 20 carbon atoms (i.e., C24-C24-C24). 2-20 alkenyl), 2 to 8 carbon atoms (i.e., C) 2-8 alkenyl), 2 to 6 carbon atoms (i.e., C) 2-6 alkenyl) or 2 to 4 carbon atoms (i.e., C) 2-4 Alkyl groups (alkenyl). Examples of alkenyl groups include: vinyl, propenyl, butadienyl (including 1,2-butadienyl and 1,3-butadienyl).

[0038] "Alkyne" refers to a group containing at least one carbon-carbon triple bond and having 2 to 20 carbon atoms (i.e., C24-C24-C24). 2-20 acetylsyl group), 2 to 8 carbon atoms (i.e., C64) 2-8 alkynyl group), 2 to 6 carbon atoms (i.e., C64) 2-6 (alkynyl group) or 2 to 4 carbon atoms (i.e., C) 2-4 Alkyl groups (alkynyl groups). The term "alkynyl" also includes alkynyl groups having one triple bond and one double bond.

[0039] "Aryl" refers to an aromatic carbocyclic group having a single ring (e.g., monocyclic) or multiple rings (e.g., bicyclic or tricyclic), including fused systems. Aryl groups, as used herein, have 6 to 20 ring carbon atoms (i.e., C64 ... 6-20 aryl), 6 to 12 carbon ring atoms (i.e., C 6-12 aryl) or 6 to 10 carbon ring atoms (i.e., C 6-10 Aryl groups. Examples of aryl groups include phenyl, naphthyl, fluorenyl, and anthracene. However, aryl groups do not include heteroaryl groups as defined below or do not overlap with heteroaryl groups in any way. If one or more aryl groups fuse with a heteroaryl ring, the resulting ring system is a heteroaryl ring.

[0040] "Cyano" refers to -CN.

[0041] “Cycloalkyl” refers to a saturated or partially unsaturated cyclic alkyl group having a single or multiple rings, including fused rings, bridged rings, and spirocyclic systems. Cycloalkyl also refers to a ring system comprising multiple carbon rings fused together, wherein one of the fused rings is an aromatic ring, but the ring system is not necessarily aromatic. Cycloalkyl groups, as used herein, have 3 to 20 ring carbon atoms (i.e., C4, C5, C6, C7, C8, C9 ... 3-20 cycloalkyl groups), 3 to 12 cyclic carbon atoms (i.e., C12+ ... 3-12 cycloalkyl groups), 3 to 10 cyclic carbon atoms (i.e., C14 and C24). 3-10 cycloalkyl groups), 3 to 8 cyclic carbon atoms (i.e., C1646-C ... 3-8 cycloalkyl groups or 3 to 6 cyclic carbon atoms (i.e., C16, C26, C36, C46, ​​C56, C6 ... 3-6 Cycloalkyl groups. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cyclohexenyl.

[0042] Unless otherwise stated, the term "halogenated" or "halogen" on its own or as part of another substituent indicates a fluorine, chlorine, bromine, or iodine atom.

[0043] "Haloalkyl" refers to an unbranched or branched alkyl group as defined above, wherein one or more hydrogen atoms are replaced by a halogen. For example, in the case where the alkyl group is substituted by more than one halogen, it may be referred to by using a prefix corresponding to the number of halogen groups attached. Dihaloalkyl and trihaloalkyl refer to alkyl groups substituted by two ("di") or three ("tri") halogen groups, which may or may not be the same halogen. Examples of haloalkyl include, but are not limited to: difluoromethyl (-CHF2), trifluoromethyl (-CF3), fluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, and 3-bromopropyl.

[0044] Unless otherwise stated, the term "heteroalkyl" on its own or in combination with another term refers to a stable straight-chain, branched, or cyclic hydrocarbon group, or a combination thereof, consisting of at least one carbon atom and at least one heteroatom selected from O, N, P, Si, and S, wherein the nitrogen and sulfur atoms may optionally be oxidized and the nitrogen heteroatom may optionally be quaternized. The heteroatoms N, O, S, P, and Si may be contained at any non-terminal position or at any position where the heteroalkyl group is linked. Two or more heteroatoms in the chain may be consecutive. Examples include, but are not limited to: -CH2-CH2-O-CH3, -CH2-CH2-O-CH2-CH2-O-CH3, -CH2-CH2-O-CH2-CH2-O-CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2-S(O)-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=N-OCH3, -CH=CH-N(CH3)-CH3, O-CH3, and -O-CH2-CH3.

[0045] "Heteroaryl" refers to an aromatic group, including groups having aromatic tautomerism or resonance structures, having a single ring, multiple rings, or multiple fused rings, and having one or more cyclic heteroatoms independently selected from N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom is optionally quaternized. Heteroaryls as used herein include 3 to 20 ring atoms (i.e., 3 to 20-membered heteroaryls), 3 to 12 ring atoms (i.e., 3 to 12-membered heteroaryls), or 5 to 10 ring atoms (i.e., 5 to 10-membered heteroaryls), and 1 to 5 heteroatoms independently selected from N, O, and S. Heteroaryls do not include aryls as defined above or do not overlap with aryls as defined above. Heteroaryls may be attached to the rest of the molecule via carbon atoms or heteroatoms. Non-limiting examples of aryl and heteroaryl groups include: phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrole, 2-pyrrole, 3-pyrrole, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, triazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isooxazolyl, 4-isooxazolyl, 5-isooxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furanyl, 3-furanyl. 2-Thienyl, 3-Thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-benzothiazolyl, purine, 2-benzimidazolyl, 5-indolyl, 1-isoquinolinyl, 5-isoquinolinyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolinyl, 6-quinolinyl, pyridin-2(1H)-one, pyridazin-3(2H)-one, pyrimidin-4(3H)-one, quinolin-2(1H)-one, pyrimidinyl, purine, pyridinyl, pyridazinyl, benzothiazolyl, and pyrazolyl. The substituents used in each of the above aryl and heteroaryl ring systems are selected from the acceptable substituents listed below.

[0046] The term "heterocyclic group" refers to the cyclic version of "heteroalkyl group". Furthermore, with respect to heterocyclic group 2, the heteroatom can occupy the position where the heterocycle is attached to the rest of the molecule. Examples of cycloalkyl groups include, but are not limited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, etc. Examples of heterocyclic alkyl groups include, but are not limited to: tetrahydropyran, 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothiophen-2-yl, tetrahydrothiophen-3-yl, 1-piperazinyl, 2-piperazinyl, etc. Examples of heterocyclic alkyl groups include, but are not limited to: glucose, mannose, allose, adroose, gulose, idose, galactose, and tarose. Examples of heterocyclic alkyl groups include, but are not limited to:

[0047] , , , , , wait.

[0048] "Hydroxyl" and "hydroxyl" are used interchangeably and refer to -OH. "Oxo" refers to an O with a double bond, written as, for example, (=O) or (O). In the presence of tautomers in a compound, hydroxyl and bridging oxygen groups are interchangeable.

[0049] "Thiol group" refers to -SH.

[0050] "Aromatic" and "heteroaryl" alone or as part of another substituent refer to divalent groups obtained from aryl and heteroaryl, respectively.

[0051] Each of the above terms (e.g., "alkyl", "heteroalkyl", "aryl" and "heteroaryl") may include substituted and unsubstituted forms of the indicated group.

[0052] As used herein, the terms “heteroatom” or “cyclic heteroatom” are intended to include oxygen (O), nitrogen (N), sulfur (S), phosphorus (P), and silicon (Si).

[0053] Certain commonly used alternative chemical names may be used. For example, divalent groups (such as divalent "alkyl", divalent "aryl", etc.) may also be referred to as "alkylene" or "alkylenyl", "arylene" or "arylenyl", respectively. Additionally, unless otherwise explicitly stated, when a combination of groups is referred to herein as a single group (e.g., aralkyl), the last group mentioned contains atoms that are attached to the remainder of the molecule.

[0054] The terms “optional” or “optionally” indicate that the event or situation subsequently described may or may not occur, and the description includes both the possibility that the event or situation occurs and the possibility that it does not occur. Additionally, the term “optionally substituted” means that any one or more hydrogen atoms on a specified atom or group may be substituted by a group other than hydrogen or may not be substituted.

[0055] Some of these compounds exist as tautomers. The tautomers are in equilibrium with each other. For example, amide-containing compounds may exist in equilibrium with imine tautomers, and carbonyl-containing compounds may exist in equilibrium with enol tautomers. Regardless of the type of tautomer shown, and regardless of the nature of the equilibrium between the tautomers, these compounds should be understood by those skilled in the art to include all tautomers. Therefore, amide-containing compounds should be understood to include their imine tautomers. Similarly, imine-containing compounds should be understood to include their amide tautomers.

[0056] Any chemical formula or structure given herein is also intended to represent the unlabeled and isotopically labeled forms of these compounds. Isotopically labeled compounds have the structure described by the chemical formula given herein, except that one or more atoms are replaced by atoms having a chosen atomic mass or mass number. Examples of isotopes that may be incorporated into the compounds of this disclosure include, but are not limited to, isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as, but not limited to: 2 H (deuterium, D) 3 H (tritium) 11 C 13 C 14 C 15 N、 18 F, 31 P, 32 P, 35 S, 36 Cl, and 125 I. The various isotope-labeled compounds disclosed herein are, for example, those incorporating a radioactive isotope (such as...) 3 H, 13 C and 14 Compounds of type C). These isotope-labeled compounds can be used in metabolic studies, reaction kinetic studies, detection or imaging techniques (such as positron emission tomography (PET) or single-photon emission computed tomography (SPECT)) including the determination of drug or substrate tissue distribution, or for the treatment of patients with radiation.

[0057] This disclosure also includes deuterated analogues of compounds of formula I, wherein one to n hydrogen atoms bonded to a carbon atom are replaced with deuterium, where n is the number of hydrogen atoms in the molecule. Such compounds may exhibit increased antimetabolite activity and, when administered to mammals (particularly humans), may be used to increase the half-life of any compound of formula I. See, for example, Foster, “Deuterium Isotope Effects in Studies of Drug Metabolism,” Trends Pharmacol. Sci. 5(12):524-527 (1984). Such compounds are synthesized using methods well known in the art, for example by using starting materials in which one or more hydrogen atoms have been replaced with deuterium.

[0058] The deuterium-labeled or substituted therapeutic compounds disclosed herein may have improved DMPK (drug metabolism and pharmacokinetics) properties, which involve distribution, metabolism, and excretion (ADME). Substitution with a heavier isotope (such as deuterium) can provide certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life, reduced dose requirement, and / or improved therapeutic index. 18 F-labeled compounds can be used in PET or SPECT studies. The isotope-labeled compounds and their prodrugs disclosed herein can generally be prepared by performing the procedures disclosed in the following embodiments and preparations, which involve replacing non-isotope-labeled reagents with readily available isotope-labeled reagents. It should be understood that, in this context, deuterium is considered a substituent in the compounds described herein.

[0059] The concentration of such heavier isotopes (specifically deuterium) can be defined by the isotope enrichment factor. In the compounds disclosed herein, any atom not specifically designated as a particular isotope is intended to represent any stable isotope of that atom. Unless otherwise stated, when a position is specifically designated as “H” or “hydrogen”, that position should be understood to contain hydrogen in its naturally occurring isotopic composition. Therefore, in the compounds disclosed herein, any atom specifically designated as deuterium (D) is intended to represent deuterium.

[0060] In some embodiments, these compounds are able to form acidic and / or basic salts due to the presence of amino and / or carboxyl groups or similar groups.

[0061] Also provided are pharmaceutically acceptable salts, hydrates, solvates, tautomers, polymorphs, and prodrugs of the compounds described herein. "Pharmaceutically acceptable" or "physiologically acceptable" refers to compounds, salts, compositions, dosage forms, and other materials that can be used to prepare pharmaceutical compositions suitable for veterinary or human pharmaceutical use.

[0062] The term "pharmaceutically acceptable salt" for a given compound refers to a salt that retains the biological potency and properties of the given compound and is not biologically or otherwise undesirable. "Pharmaceutically acceptable salt" or "physiologically acceptable salt" includes, for example, salts formed with inorganic acids and salts formed with organic acids. Additionally, if the compound described herein is obtained as an acid addition salt, the free base can be obtained by alkalizing a solution of the acid salt. Conversely, if the product is a free base, the addition salt, specifically a pharmaceutically acceptable addition salt, can be prepared according to conventional procedures for preparing acid addition salts from basic compounds by dissolving the free base in a suitable organic solvent and treating the solution with an acid. Those skilled in the art will recognize the various synthetic methods that can be used to prepare non-toxic, pharmaceutically acceptable addition salts. Pharmaceutically acceptable acid addition salts can be prepared from inorganic and organic acids. Salts obtained from inorganic acids include: hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. Salts derived from organic acids include: acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid. Similarly, pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases. Salts derived from inorganic bases include, to name only, salts of sodium, potassium, lithium, ammonium, calcium, and magnesium. Salts obtained from organic bases include, but are not limited to: salts of primary, secondary, and tertiary amines, such as alkylamines (i.e., NH2(alkyl), dialkylamines (i.e., HN(alkyl)2), trialkylamines (i.e., N(alkyl)3), alkenylamines (i.e., NH2(alkenyl)), dienylamines (i.e., HN(alkenyl)2), trienylamines (i.e., N(alkenyl)3), mono-, di-, or tricycloalkylamines (i.e., NH2(cycloalkyl), HN(cycloalkyl)2, N(cycloalkyl)3), mono-, di-, or triarylamines (i.e., NH2... ( Aryl, HN(aryl)2, N(aryl)3), or mixed alkyl, alkenyl, cycloalkyl, and / or aryl amines. Specific examples of suitable amines include, by way of example only: diisopropylamine, triethylamine, diethylamine, tri(isopropyl)amine, tri(n-propyl)amine, ethanolamine, 2-dimethylaminoethanol, piperazine, piperidine, morpholine, N-ethylpiperidine, etc.

[0063] The term "substituted" means that any one or more hydrogen atoms on a specified atom or group are replaced by one or more substituents other than hydrogen, provided that the valence of the specified atom is not exceeded. Unless otherwise stated, one or more substituents can be any substituents provided herein or a combination thereof. Polymers or similar indeterminate structures obtained by defining substituents with an unlimited number of additional substituents (e.g., a substituted aryl group having a substituted alkyl group, wherein the substituted alkyl group itself is further replaced by a substituted aryl group substituted with a substituted heteroalkyl group, etc.) are not intended to be included herein.

[0064] As used herein, "pharmaceutically acceptable carrier" or "pharmaceutical acceptable excipient" includes any and all solvents, dispersion media, coating materials, antibacterial and antifungal agents, isotonic agents, and absorption delay agents. The use of such media and agents for pharmaceutically active substances is well known in the art. Their use in pharmaceutical compositions is conceivable unless any conventional media or agent is incompatible with the active ingredient. Complementary active ingredients may also be incorporated into the composition.

[0065] A solvate is formed through the interaction of a solvent and a compound. Solvates of salts of the compounds described herein are also provided. Hydrates of the compounds described herein are also provided.

[0066] "Produce drug" refers to any compound that, when administered to a biological system, generates a parent compound through spontaneous chemical reactions, enzyme-catalyzed chemical reactions, photolysis, and / or metabolic chemical reactions. Therefore, a produce drug is a covalently modified analogue or potential form of a biologically active parent compound. In some embodiments, the parent compound is compound 7.

[0067] compound

[0068] This disclosure provides compounds that can be used for the detection and treatment of neurological diseases and conditions. The compounds disclosed herein can be used for the detection of a patient's disease or condition, screening to identify patients at risk of developing a disease or condition, diagnosis of a disease or condition, or monitoring of a disease or condition. The compounds disclosed herein can act as prodrugs; therefore, the compounds described herein are intended to include metabolites and parent compounds formed upon administration to a patient.

[0069] Due to the poor water solubility and large particle size of compounds (such as compound 7), prodrug derivatives of these compounds have been studied. It has been found that the phosphate prodrug of compound 7 exhibits advantages in solubility and reduced crystallinity. Specifically, it was found that water solubility increased upon the addition of a phosphate ester functional group to compound 7. It was also found that the phosphate functional group is cleaved by phosphatases in vivo, leading to the delivery of the parent compound.

[0070] The compound may be a compound of formula I or a pharmaceutically acceptable salt thereof:

[0071] I

[0072] Where R 1 It is selected from:

[0073] , , , , , , , , , , , , , , , , , , ,and .

[0074] Each X is independently O or S;

[0075] Each R 11 It is independently selected from hydrogen and C 1-10 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 6-10 Aryl, 5- to 10-membered heteroaryl, and 4- to 10-membered heterocyclic groups; wherein the alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, and heterocyclic groups are optionally surrounded by 1-4 R groups. 21 Replace; or each XR 11 Independently is -XP(X)(R1 2 )2;

[0076] Each R 12 It is independently selected from hydroxyl, mercapto, -XP(X)(R1) 3 2. C 1-10 Alkyl, -OC 1-10 Alkyl and -SC 1-10 alkyl;

[0077] Each R 13 It is independently selected from hydroxyl, mercapto, C1-10 alkyl, -OC 1-10 Alkyl and -SC 1-10 alkyl;

[0078] Each R 21It is independently selected from halogens, hydroxyl groups, mercapto groups, -NO2, -N3, cyano groups, and C. 1-10 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 1-8 Haloalkyl, C 6-10 Aryl, 5- to 10-membered heteroaryl, 4- to 10-membered heterocyclic, -OC 1-10 Alkyl, -OC 2-6 alkenyl, -OC 2-6 alkynyl group, -OC 3-10 cycloalkyl, -OC 1-8 Haloalkyl, -O-aryl, -O-heteroaryl, -O-heterocyclic, -NH2, -NH(R) 31 ), -N(R 31 )2、-C(O)(R 31 -C(O)O(R) 31 ), -C(O)OH, -C(O)NH2, -C(O)NH(R 31 -C(O)N(R) 31 )2、-NHC(O)(R 31 ), -NHC(O)O(R 31 ), -NHC(O)NH(R 31 ), -S(R 31 -NHS(O) y (R 31 -N(C1-10 alkyl)S(O) y (R) 31 ), -S(O) y N(R 31 )2、-S(O)NH(R 31 ), and -S(O) y (R 31 );

[0079] Each R 31 It is independently selected from C 1-10 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 1-8 Halogenated alkyl, aryl, heteroaryl, and heterocyclic groups; and

[0080] Each y is independently 1 or 2.

[0081] In some implementation schemes, R 1 yes .

[0082] In some implementations, each X is independently O.

[0083] In some implementation schemes, R 1 yes , where X 1 X 2 and X 3 Each can be either O or S independently.

[0084] In some implementation schemes, R 1 yes .

[0085] In some implementation schemes, each R 11 It is hydrogen on its own.

[0086] In some implementation schemes, R 1 yes .

[0087] In some embodiments, the compound is a monobasic salt.

[0088] In some embodiments, the compound is a disalt.

[0089] In some embodiments, the compound is a salt selected from sodium, potassium, lithium, ammonium, calcium, magnesium, or zinc salts.

[0090] In some embodiments, the compound is a sodium salt.

[0091] In some embodiments, the compound is a potassium salt.

[0092] In some embodiments, the compound is an ammonium salt. In some embodiments, the compound is a diammonium salt.

[0093] In some implementations, the compound of formula I is

[0094]

[0095] Or it may be pharmaceutically acceptable to use salt.

[0096] In some embodiments, the compound is compound 7:

[0097]

[0098] Or it may be pharmaceutically acceptable to use salt.

[0099] In some embodiments, the compound is compound 8:

[0100]

[0101] Or it may be pharmaceutically acceptable to use salt.

[0102] In some embodiments, the compound is compound 9:

[0103]

[0104] Or a pharmaceutically acceptable salt. In some embodiments, the salt is a diammonium salt. In some embodiments, the compound is compound 10:

[0105] .

[0106] General Synthesis

[0107] The compounds of this disclosure can be prepared using the methods disclosed herein and their conventional modifications, as will be apparent from the disclosure herein and methods well known in the art. In addition to the teachings herein, conventional and well-known synthetic methods can be used. The synthesis of typical compounds of formula (I) (e.g., compounds having the structure described by formula (I) or compounds disclosed herein, or their pharmaceutically acceptable salts) can be carried out using methods as described in the examples and methods known in the art.

[0108] Typical embodiments of the compounds according to the invention can be synthesized using the reaction diagrams and / or examples described below. Given the description herein, it will be apparent that the scheme can be modified by substituting other materials having similar structures to obtain correspondingly different products. The description of the following synthesis provides examples of how the steps can be varied to yield the desired product. The group designations (e.g., R1) used in the reaction diagrams herein are for illustrative purposes only and, unless otherwise stated, do not necessarily match the designations used elsewhere to describe compounds of formula (I) or aspects or fragments thereof, in terms of name or function.

[0109] It should be understood that, unless otherwise specified, other process conditions may be used given the process conditions (i.e., reaction temperature, time, molar ratio of reactants, solvent, pressure, etc.). Optimal reaction conditions may vary depending on the specific reactants or solvents used, but such conditions can be determined by those skilled in the art through conventional optimization procedures. Furthermore, it will be apparent to those skilled in the art that conventional protecting groups are necessary to prevent certain functional groups from undergoing undesirable reactions. Suitable protecting groups for various functional groups, and suitable conditions for protecting and deprotecting specific functional groups, are well known in the field. For example, many protecting groups are described in, for instance, TW Greene and GM Wuts (1999), Protecting Groups in Organic Synthesis, 3rd Edition, Wiley, New York, and the references cited therein.

[0110] The materials and reagents used in the following reactions are generally known compounds, or can be prepared by known methods or obvious modifications thereof. For example, many starting materials are available from suppliers such as Aldrich Chemical Co. (Milwaukee, Wisconsin, USA). Other compounds can be prepared by the processes described in the following standard reference texts or by obvious modifications thereof: *Fieser and Fieser's Reagents for Organic Synthesis*, Volumes 1-15 (John Wiley and Sons, 1991); *Rodd's Chemistry of Carbon Compounds*, Volumes 1-5 and *Supplementals* (Elsevier Science Publishers, 1989); *Organic Reactions*, Volumes 1-40 (John Wiley and Sons, 1991); *March's Advanced Organic Chemistry* (John Wiley and Sons, 5th Edition, 2001); and *Larock's Comprehensive Organic Transformations* (VCH Publishers). Inc., 1989.

[0111] Procedure 1 illustrates an exemplary synthetic route for the synthesis of compounds provided herein (e.g., compounds of Formula I). ​​The desired R can be synthesized by first providing compound 7 and then using suitable conditions (e.g., nucleophilic substitution). 1 Substituents are used to prepare compounds of formula I, such as compound 11, or other formulas or compounds disclosed herein. In process 1, R 1 As defined in this paper, Y is a suitable leaving basis.

[0112] Process 1

[0113]

[0114] In process 1, compound 7 is reacted with compound 12 under nucleophilic substitution conditions (e.g., using a base) in a suitable solvent (e.g., THF, DMF, etc.) with compound 12 in an inert atmosphere to provide compound 11. In some cases, compound 7 is deprotonated with a base (e.g., sodium hydride or butyllithium) and then contacted with compound 12. The reaction can be carried out at a temperature of about -78 to 0°C for about 5 minutes to about 1 hour, or at a temperature of about 0 to 50°C for about 1 hour to about 12 hours. When the reaction is substantially complete, product compound 11 is isolated by conventional methods.

[0115] Detectable target proteins

[0116] The compounds described herein can be used for the detection or treatment of neurological diseases or conditions. In this regard, the compounds described herein can be prodrugs. Therefore, the compounds described herein can be converted into parent compounds via chemical or enzymatic pathways, and the parent compounds can be used accordingly for the detection or treatment of neurological diseases or conditions.

[0117] Many neurological disorders, including neurodegenerative diseases and injury-related disorders, can be detected using the compounds and methods described herein. Neurological disorders or conditions are characterized by the accumulation of certain peptides, proteins, or proteins (described herein as detectable proteins). Detectable proteins or their accumulated mass may include, for example, amyloid-β protein or phosphorylated tau protein. As described herein, amyloid-β protein or phosphorylated tau protein can be detected by contact with a compound. Typically, the compounds and methods described herein can be used to detect amyloid-β protein or phosphorylated tau protein or their accumulated mass in a patient's tissue or sample. This presence of amyloid-β protein or phosphorylated tau protein can be detected using a compound that binds to amyloid-β protein or phosphorylated tau protein, and then the binding can be detected.

[0118] Amyloid β protein (Aβ) is typically a polypeptide containing about 40 amino acid residues, such as about 36-43, 39-43, or 40-42 amino acid residues. Isoforms include Aβ(1-40) and Aβ(1-42). In some embodiments, Aβ is Aβ(1-42). Aβ is generally believed to be produced by enzymatic cleavage of the larger precursor protein β-amyloid precursor protein (APP), which is encoded by a gene on human chromosome 21. APP aberrations include: NP_000475.1, NP_001129488.1, NP_001129601.1, NP_001129602.1, NP_001129603.1, NP_001191230.1, NP_001191231.1, NP_001191232.1, NP_958816.1, and NP_958817.1. Aβ is generally believed to be produced by the action of secretases β and γ on APP. Aβ has been found in deposits (e.g., plaques) in the brains of individuals with Alzheimer's disease. Aβ is thought to be involved in the pathogenesis of neurological diseases. Aβ is also considered to be toxic to nerve cells.

[0119] The proteins detected by the compounds of this disclosure include: amyloid β peptide (Aβ), viral peptide (PrP), α-synuclein, IAPP (amyloid), huntingtin, calcitonin (ACal), atrial natriuretic peptide (AANF), apolipoprotein A1 (ApoA1), serum amyloid A (SAA), medroxyprolactin (AMed), prolactin (APro), thyroxine transporter (ATTR), lysozyme (ALys), β2-microglobulin (Aβ2M), colloidin (AGel), corneal epithelial protein (Aker), cystatin (ACys), immunoglobulin light chain AL (AL), S-IBM, or superoxide dismutase. In some embodiments, the detected amyloid peptide is Aβ peptide, viral peptide, α-synuclein, or superoxide dismutase.

[0120] “Microtubule-associated protein tau,” “MAPT,” “tau protein,” or “tau” are a class of proteins that stabilize microtubules during assembly and disassembly, and are classified as microtubule-associated proteins (MAPs). Tau isoform sequences include: NP_001116538.2, NP_001116539.1, NP_001190180.1, NP_001190181.1, NP_005901.2, NP_058518.1, NP_058519.3, and NP_058525.1. Tau proteins are important in the stability and assembly of microtubules and accordingly affect intraneuronal transmission of cargo. Tau may also participate in signaling pathways through interaction with actin at its acidic N-terminus, thereby protruding from the microtubule to promote neurite growth and stability during neural development. The tau proteins described herein may include any isoform or any combination of isoforms. MAPT transcripts are expressed differently in the nervous system, depending on the stage of neuronal maturation and neuron type. Mutations in the MAPT gene have been associated with a variety of neurological disorders, such as Alzheimer's disease, Pick's disease, frontotemporal dementia, cortical-basal degeneration, and progressive supranuclear palsy. Tau proteins may or may not contain post-translational modifications. The tau protein family is characterized by a shared N-terminal segment, a ~50-amino acid sequence inserted into the N-terminal segment that is developmentally regulated in the brain, a characteristic tandem repeat region consisting of 3 or 4 tandem repeats, and a C-terminal tail.

[0121] The human tau gene is located at 17q21 on the long arm of chromosome 17. It is generally believed to contain 16 exons, with exon 21 being part of the promoter. The primary tau transcript contains 13 exons, and exons 4A, 6, and 8 are not transcribed in humans. Exons 21 and 14 are transcribed but not translated. Exons 1, 4, 5, 7, 9, 11, 12, and 13 are constitutive, and either exons 2, 3, and 10 are spliced, resulting in six different mRNAs translated in six different tau isoforms. These isoforms differ in the presence or absence of one or two 29-amino acid repeats (ON, 1N, or 2N) encoded by exons 2 and 3 in the N-terminal region, or three microtubule-binding repeats (R1, R3, and R4) or four repeat regions (R1-R4) in the C-terminal region. The fourth microtubule-binding domain is encoded by exon 10. Six tau protein isoforms are known to exist in human brain tissue: the (2+3+10+) isoform (441 amino acids), the (2+3+10-) isoform (410 amino acids), the (2+3+10-) isoform (412 amino acids), the (2+3-10-) isoform (381 amino acids), the (2-3-10+) isoform (383 amino acids), and the (2-3-10-) isoform (352 amino acids). This tau can be a mutant tau. This mutation could be the FTDP-17 mutation. Examples of mutations include: G272V, N279K, N296, P201L, P301S, G303V, S305N, L315R, S320F, P332L, V337M, E342V, S352L, K369I, G389R, R5H, R5L, K257T, I260V, L266V, G272V, delK280, N296H, N296N, delN296, P301L, P301S, K317M, G335V, Q336R, R406W, and R427M.

[0122] Phosphorylated tau protein or "phosphorylated tau" is a tau protein having at least one amino acid residue modified with a phosphate group. Tau is generally considered to comprise up to 85 phosphorylated amino acid residues. Typically, the phosphate group is a post-translational modification and can be bound to the side chain of an amino acid residue. The phosphorylated amino acid residue can be, for example, a serine (S), threonine (T), or tyrosine (Y) residue, or a combination thereof. Phosphorylated tau protein may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, at least 20, at least 30, at least 40, or at least 50 moles of phosphate ester per mole of protein. Phosphorylated tau protein may contain at least 3 moles of phosphate ester per mole of protein. Phosphorylated tau proteins may include one or more phosphorylated amino acid residues selected from Thr39, Ser46Pro, Thr50Pro, Thr69Pro, Thr153Pro, Thr175Pro, Thr181Pro, Ser198, Ser199, Ser202Pro, Thr205Pro, Ser208, Ser210, Thr212Pro, Ser214, Thr217Pro, Thr231Pro, Ser235Pro, Ser237, Ser241, Ser262, Ser285, Ser305, Ser324, Ser352, Ser356, Ser396Pro, Ser400, Thr403, Ser404Pro, Ser409, Ser412, Ser413, Ser416, and Ser422Pro. Phosphorylated tau proteins may include phosphorylated Ser422. Phosphorylated tau proteins as described herein may be deposited or non-deposited. Phosphorylated tau proteins as described herein may be soluble. Tau proteins or phosphorylated tau proteins may be triplicate tau, tetralicate tau, or a combination thereof. In some embodiments, tau proteins or phosphorylated tau proteins may comprise a mixture of triplicate and tetralicate tau, wherein tetralicate tau is more common. In some embodiments, tau proteins or phosphorylated tau proteins may comprise a mixture of triplicate and tetralicate tau, wherein triplicate tau is more common. Phosphorylated tau proteins may be filamentous, for example, as neurofibrillary tangles (NFTs). NFTs can be found in the dendritic chambers of neurons.

[0123] "Contact" is used in its general sense and refers to the process of bringing at least two different kinds (e.g., chemical compounds containing biomolecules, or cells) close enough to interact. The term "contact" can include allowing two species to react or physically touch, wherein said two kinds can be compounds, biomolecules, proteins, or enzymes as described herein. In some embodiments, contact includes causing a compound described herein to interact with a protein (e.g., Aβ protein or phosphorylated tau protein) or enzyme.

[0124] Therefore, according to some embodiments of this disclosure, a method is provided for determining whether a patient suffers from a neurological disease or condition. This method requires detecting the presence or cumulative mass of amyloid-β protein or phosphorylated tau protein in the patient's tissue or sample by contacting the patient's tissue or sample with the compound described herein. Contact can be in vivo or ex vivo. Contact can be performed by administering the compound, for example, locally or intravenously to the patient.

[0125] In another embodiment, a method for preparing a patient for diagnosis of a neurological disease or condition is provided, the method comprising administering a compound described herein to the patient and detecting its binding to amyloid β-protein or phosphorylated tau protein or amyloid protein, or their accumulated mass. The compound may be administered intravenously. Once the compound has been administered to the patient, its binding and / or the binding of the parent compound to amyloid β-protein or phosphorylated tau protein, or their accumulated mass, can be detected by any method, including those described herein. In some embodiments, binding indicates the likelihood that the patient has a neurological disease or condition.

[0126] Neurological diseases and their treatment

[0127] In some embodiments, this disclosure provides a method for determining the presence or absence of a neurological disease or condition in a patient. In some embodiments, the method includes administering an effective amount of a compound or pharmaceutical composition thereof described herein to the patient. The compound may be a compound of formula I. In some embodiments, the compound is compound 7, compound 8, or compound 9. In some embodiments, the compound is compound 7, compound 8, compound 9, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is compound 10. In some embodiments, a method for determining whether a patient suffers from a neurological disease or condition is provided, the method comprising administering a compound or pharmaceutical composition thereof described herein to the patient. In some embodiments of the method, the compound is administered intravenously. In some embodiments of the method, the compound is administered to the patient's eye. In some embodiments, the neurological disease or condition is a disease or condition characterized by protein deposition or protein misfolding.

[0128] This document also provides methods for determining whether a patient has a neurological disease or condition, including detecting the presence or cumulative mass of amyloid β-protein or phosphorylated tau protein in the patient's tissue or sample, including contacting the tissue or sample with a compound described herein. The compound may be a compound of formula I. In some embodiments, the compound is compound 7, compound 8, or compound 9. In some embodiments, the compound is compound 7, 8, 9, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is compound 10: the contact may be in vivo. The tissue may be ocular tissue. The sample may be a urine sample.

[0129] In some embodiments, the neurological disease or condition is selected from age-related diseases or conditions, genetic diseases or conditions, injury-related diseases or conditions, and mental illnesses or conditions. In some embodiments, age-related diseases or conditions are selected from Parkinson's disease, vascular dementia, and amyotrophic lateral sclerosis (ALS); genetic diseases or conditions are Down syndrome; injury-related diseases or conditions are selected from traumatic brain injury and chronic traumatic encephalopathy; and mental illnesses or conditions are selected from schizophrenia and depression. The neurological disease or condition may be tau proteinosis. In some embodiments, the neurological disease or condition is Alzheimer's disease or traumatic brain injury (TBI).

[0130] Neurological disorders or conditions can be tau proteinopathies. Cranioencephalopathy is a class of neurological diseases associated with the pathological deposition of tau protein in neurofibrillary or glial fibrillary tangles in the human brain. Tangles can form through hyperphosphorylation of tau, leading to the dissociation of tau protein from microtubules and the formation of insoluble deposits. These deposits of hyperphosphorylated tau protein can also be referred to as paired helical fibers. The exact mechanism of tangle formation is not fully understood, and whether tangles are the primary pathogenic factor or play a greater peripheral role remains controversial. Tau proteinopathies have been found in many neurological diseases, such as post-traumatic degeneration, infection, metabolic diseases, and motor neuron degeneration. The spatial distribution, transient appearance, and structural changes of tau protein vary across different neurological diseases. Patients with Alzheimer's disease (AD) have twisted, hyperphosphorylated, and non-periodic tau monofilaments, while patients with progressive hypernuclear palsy and frontotemporal dementia (FTD) often have only straight tau monofilaments. Tau proteinopathies often overlap with conuclear proteinopathies, possibly due to interactions between synuclein and tau proteins. Non-Alzheimer's diseases are sometimes grouped together with "Pick syndrome" because they are associated with frontotemporal dementia or frontotemporal degeneration. A hallmark of tau hyperphosphorylation is tau pS422. Chronic traumatic encephalopathy (CTE) is associated with recurrent mild traumatic brain injury (mTBI) and shares many similarities with Taopathies, including hyperphosphorylation and deposition of tau proteins, such as neurofibrillary tangles (NFTs).

[0131] In some implementations, the neurological disease or condition is selected from primary age-related tau protein disease (PART), senile dementia with neurofibrillary tangles, chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), frontotemporal dementia and Parkinson's disease with chromosome 17 (FTDP-17), Lytico-bodig disease (Guam Parkinson's disease-dementia), ganglioglioma, gangliocytoma, meningioma, cerebral Parkinson's disease, subacute sclerosing panencephalitis (SSPE), lead encephalopathy, tuberous sclerosis, pantothenic acid-related neurodegeneration, and lipofuscin deposition disease.

[0132] Neuronal diseases or conditions can be neurodegenerative diseases or conditions. In some implementations, the neurological disease or condition is Alzheimer's disease. Alzheimer's disease can be classified as secondary tau proteinopathy. Alzheimer's disease is characterized by memory loss in the early stages of the disease. Neurofibrillary tangles are an early description of Alzheimer's disease. When tau becomes hyperphosphorylated, the protein dissociates from the microtubules of the axon. Tau then becomes misfolded and begins to deposit, which can form neurofibrillary tangles (NFTs). Microtubules are also unstable when tau is dissociated, and the combination of neurofibrillary tangles and unstable microtubules leads to disruption of processes such as axonal transport and neural communication. The extent to which NFTs are involved in Alzheimer's disease is defined by Braak stages. Braak stages I and II are used when NFT involvement is primarily confined to the transgut region of the brain. When limbic regions such as the hippocampus are involved, Alzheimer's disease progresses to stages III and IV; when extensive involvement of the neocortex is observed, it progresses to stages V and VI. Due to the presence of senile plaques, Alzheimer's disease is also classified as amyloidosis. Furthermore, certain Apoε4 carriers have a greater risk of developing Alzheimer's disease. APOε4 is generally considered less effective than other isoforms in clearing Aε, and is therefore associated with greater amyloid burden, tau phosphorylation, synaptic toxicity, and decreased synaptic density. Having experienced traumatic brain injury (TBI) is another risk factor for AD, and studies have shown that those who have experienced TBI have a significantly increased risk of developing AD.

[0133] As the disease progresses, symptoms include confusion, long-term amnesia, incoherent speech, vocabulary loss, aggression, irritability, and / or mood swings. In later stages of the disease, loss of bodily function is present. Patients with Alzheimer's disease (AD) exhibit a number of characteristic neuropathies, such as increased oxidative stress, mitochondrial dysfunction, synaptic dysfunction, disruption of calcium homeostasis, deposition of senile plaques and neurofibrillary tangles, and brain atrophy. AD-related symptoms include: Alzheimer's disease type 2 (SDAT), frontotemporal dementia (FTD), vascular dementia, mild cognitive impairment (MCI), and age-related memory impairment (AAMI). In some implementations, determining whether a patient has Alzheimer's disease involves detecting the presence of phosphorylated tau protein in the patient's tissues or samples, wherein this detection involves contacting the phosphorylated tau protein with the compounds described herein.

[0134] In some embodiments, the neurological disease or condition is frontotemporal degeneration (FTLD) (e.g., FTLD-tau, FTLD-TDP, or FTLD-FUS). In some embodiments, the neurological disease or condition is frontotemporal dementia. In some embodiments, the neurological disease or condition includes memory loss. In some embodiments, the neurological disease or condition is age-related memory loss. In some embodiments, the neurological disease or condition is type A FTLD-TDP. In some embodiments, the neurological disease or condition is type B FTLD-TDP. In some embodiments, the neurological disease or condition is type C FTLD-TDP. In some embodiments, the neurological disease or condition is type D FTLD-TDP.

[0135] In some embodiments, the neurological disease or condition is Parkinson's disease. In some embodiments, the neurological disease or condition is Parkinson's dementia. In some embodiments, the neurological disease or condition is associated with (e.g., characterized by) the accumulation of amyloid plaques. In some embodiments, the patient with the neurological disease or condition has suffered traumatic brain injury before, during, or after the onset of neuronal disease. In some embodiments, the neurological disease or condition includes neuronal damage. Neuronal damage may include atrophy or other reduction in the effective function of neurons. For example, Alzheimer's disease is known to manifest as neuronal damage, particularly cortical neurons, such as hippocampal neurons and neurons near the hippocampus.

[0136] In some implementations, the neurological disease or condition is traumatic axonal injury (TAI), traumatic encephalopathy (TBD), dementia (e.g., common dementia), frontal lobe dementia, Parkinson's disease associated with chromosome 17 (FTDP-17), primary age-related tau proteinosis (PART), neurofibrillary tangles-predominant Alzheimer's disease, progressive supranuclear palsy (PSP), corticobasal degeneration, Lytico-Bodig disease (Guam's Parkinson-dementia complex), ganglioglioma, gangliocytoma, meningioma, encephalomyelitis, encephalomyelitis, tuberous sclerosis, pantothenic kinase-related neurodegeneration, lipofuscinosis, Pick's disease, corticobasal degeneration, aerobatic dysplasia (AGD), or corticobasal degeneration.

[0137] Neurological disorders or impairments can be injury-related conditions such as traumatic brain injury (TBI) or chronic traumatic encephalopathy (CTE). TBI is a chronic condition defined as damage to the brain caused by external forces such as impacts, blows, jolting, rapid acceleration or deceleration, or bullet penetration. Damage leading to TBI can result in a diminished or altered state of consciousness, leading to temporary or permanent impairment of cognitive, sensorimotor, and psychosocial functioning. CTE is a progressive degenerative disease found in individuals with recurrent traumatic brain injuries, including head impacts that do not cause TBI symptoms. Physical aspects of CTE include: brain atrophy, atrophy of the frontal and temporal lobes, enlarged ventricles, and atrophy of the hippocampus, thalamus, brainstem, and cerebellum. Individuals with CTE may experience symptoms such as dementia, memory loss, aggression, confusion, depression, and suicidal ideation, which may occur years after the injury.

[0138] Neurological diseases or conditions can affect the eyes, such as glaucoma, high intraocular pressure, macular degeneration, diabetic retinopathy, age-related macular degeneration (AMD), or retinitis pigmentosa.

[0139] In some implementations of this method, the neurological disease or condition is prion disease. Prions are fatal, contagious neurological diseases, the most prominent of which is Creutzfeldt-Jakob disease (CJD). CJD is sometimes called a “giant imitator” because it causes symptoms found in many other neurological disorders. Common symptoms of CJD include various neurological and psychiatric indications, including behavioral changes, confusion, cognitive impairment, motor problems, and visual disturbances. CJD is difficult to diagnose, and current practice relies on cerebrospinal fluid testing. Interestingly, recent reports have shown the presence of viral proteins in the post-mortem retinal tissue of CJD patients.

[0140] Therefore, a method for detecting prion deposits in a patient is provided. In some embodiments, the detection is performed in the retina. In some embodiments, a method for detecting viral deposits in the retina of a patient is provided, wherein the patient may or may not exhibit clinical manifestations of an associated disease or condition such as Creutzfeldt-Jakob disease (e.g., behavioral changes, confusion, cognitive impairment, motor problems, visual impairment, kyphosis, ataxia, toe walking, etc.).

[0141] Cerebral amyloid angiopathy (CAA) is an age-related disease characterized by the deposition of amyloid proteins within the walls of blood vessels in the brain. These deposits form in the cortical and pia mater arteries, leading to an increased risk of spontaneous intracranial hemorrhage, ischemic lesions, and progressive dementia in older adults. Because the symptoms resemble transient ischemic attacks or “mini-strokes,” physicians often miss the diagnosis of CAA. Diagnosis can be more complex, as CAA is found in up to 90% of Alzheimer’s disease patients. Several peptide-forming amyloid proteins can contribute to CAA. Of these, amyloid-β (Aβ) is by far the most prevalent form. Deposits of Aβ have been detected in the small arteries, capillaries, and arterial walls of brain tissue in individuals with severe CAA. It has been determined that the Aβ42 isoform is preferentially present in the parenchymal plaques of Alzheimer’s disease patients, while the Aβ40 isoform is more densely packed in the cerebral vascular wall deposits of CAA. In some embodiments of this approach, the neurological disease or condition is cerebral amyloid angiopathy (CAA). Cerebral amyloid angiopathy (CAA) is an age-related disease characterized by the deposition of amyloid protein in the walls of cerebral blood vessels.

[0142] Therefore, in some embodiments, a method for detecting Aβ40 in a patient is provided. In some embodiments, the detection is performed in the retina. In some embodiments, the detection distinguishes between Aβ and CAA-associated atypical forms (Aβ40) and Alzheimer's disease (Aβ42).

[0143] Examples of neuronal diseases that can be treated with the compounds or methods described in this article include: Alexander's disease, Alper's disease, Alzheimer's disease, depression, perinatal asphyxia, Parkinson's dementia ("PD dementia"), amyotrophic lateral sclerosis, ataxia-telangiectasia, Batten disease (also known as Spielmeyer-Vogt-Sjogren-Batten disease), spongiform encephalopathy (e.g., bovine spongiform encephalopathy (mad cow disease), Kuru disease, Creutzfeldt-Jakob disease), fatal familial insomnia, Canavan disease, Cockayne syndrome, corticobasal degeneration, Fragile X syndrome, frontotemporal dementia, Gerstmann-Straussler-Scheinker syndrome, and Huntington's disease. Diseases, HIV-related dementia, Kennedy's disease, Krabbe's disease,Lewy body dementia, Machado-Joseph disease (spinocerebellar ataxia type 3), multiple sclerosis, multiple system atrophy, narcolepsy, spirochetal neuropathy, Parkinson's disease, Pelizaeus-Merzbacher disease, Pick's disease, amyotrophic lateral sclerosis, prions, Refsum's disease, Sandhoff's disease, Schilder's disease, subacute spinal cord degeneration secondary to pernicious anemia, schizophrenia, spinocerebellar ataxia (multiple types with different characteristics), spinal muscular atrophy, Steele-Richardson-Olszewski disease, Tabess disease. Dorsalis, drug-induced Parkinson's disease, progressive supranuclear palsy, corticobasal degeneration, multiple system atrophy, idiopathic Parkinson's disease, autosomal dominant Parkinson's disease, familial type 1 (PARK1), Parkinson's disease 3, autosomal dominant Lewy body (PARK3), Parkinson's disease 4, autosomal dominant Lewy body (PARK4), Parkinson's disease 5 (PARK5), Parkinson's disease 6, autosomal recessive early-onset (PARK6), Parkinson's disease 2, autosomal recessive juvenile (PARK2), Parkinson's disease 7, autosomal recessive early-onset (PARK7), Parkinson's disease 8 (PARK8), Parkinson's disease 9 (PARK9), Parkinson's disease 10 (PARK10), Parkinson's disease 11 (PARK11), Parkinson's disease 12 (PARK12), Parkinson's disease 13 (PARK13), and mitochondrial Parkinson's disease.

[0144] After a neurological disorder or condition is diagnosed in a patient, certain procedures can be provided to treat or improve the symptoms of the disorder or condition, or to slow or stop its progression. Once a neurological disorder or condition is diagnosed, the methods described in this article can also be used to monitor its progression. Once diagnosed, the attending physician can also recommend other treatments as described in this article.

[0145] "Treatment" or "treatment" is a method of achieving a beneficial or desired outcome (including clinical outcomes). A beneficial or desired clinical outcome may include one or more of the following: (a) suppressing a disease or condition (e.g., reducing one or more symptoms caused by the disease or condition, and / or reducing the severity of the disease or condition); (b) improving, slowing, or preventing the development of one or more clinical symptoms associated with the disease or condition (e.g., stabilizing the disease or condition, preventing or delaying the worsening or progression of the disease or condition and / or preventing or delaying the spread of the disease or condition (e.g., metastasis); and / or (c) alleviating the disease, i.e. causing the remission of clinical symptoms (e.g., improving the disease state, partially or completely alleviating the disease or condition, enhancing the efficacy of another medication, delaying the progression of the disease, improving quality of life, and / or prolonging survival).

[0146] "Prevention" or "preventing" refers to the treatment of any disease or condition that prevents the development of clinical symptoms. In some implementations, the compound may be administered to subjects (including humans) who are at risk of developing the disease or condition or who have a family history of the disease or condition.

[0147] "Patient" refers to an animal, such as a mammal (including a human), that has been or will be the subject of treatment, observation, or experimentation. The methods described herein can be used for human therapeutics and / or veterinary applications. In some embodiments, the patient is a mammal. In one embodiment, the patient is a human.

[0148] The term "therapeutic effective amount" or "effective amount" as used herein for a compound or its pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug, or deuterated analogue thereof means an amount sufficient to treat and provide a therapeutic benefit (e.g., symptom improvement or slowing disease progression) when administered to a subject. For example, a therapeutic effective amount may be an amount sufficient to alleviate symptoms of the disease or symptom of TBI. The therapeutic effective amount can vary depending on the subject and the disease or symptom being treated, the subject's weight and age, the severity of the disease or symptom, and the route of administration, and the therapeutic effectiveness can be readily determined by one of ordinary skill in the art.

[0149] The methods described herein can be applied to cell populations, either in vivo or in vitro. "In vivo" means within a living individual, such as an animal or human. In this context, the methods described herein can be used therapeutically in an individual. "In vitro" means outside a living individual. Examples of in vitro cell populations include in vitro cell cultures and biological samples comprising liquid or tissue samples obtained from an individual. Such samples can be obtained by methods well known in the art. Exemplary biological fluid samples include blood, cerebrospinal fluid, urine, and saliva. In this context, the compounds and compositions described herein can be used for a variety of purposes, including therapeutic and experimental purposes. For example, the compounds and compositions described herein can be used in vitro for a given indication, cell type, individual, and other parameters to determine the optimal timing and / or dosage of administration of the disclosed compounds. Information gathered from this use can be used for experimental purposes or in clinical settings to establish in vivo treatment protocols. Other in vitro uses of the compounds and compositions described herein may be suitable are described below or will become apparent to those skilled in the art. Selected compounds can be further characterized to examine safe or tolerable doses in human or non-human subjects. Such properties can be examined using methods commonly known to those skilled in the art.

[0150] Detection of target proteins

[0151] This document provides a method for diagnosing a neurological disease or condition in a patient, comprising administering a compound described herein to the patient's tissues. The compound may be a compound of formula I. In some embodiments, the compound is compound 7, compound 8, or compound 9. In some embodiments, the compound is compound 7, compound 8, compound 9, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is compound 10. The method may include detecting the binding of the compound to a detectable target protein, such as amyloid β-protein or phosphorylated tau protein, and / or the binding of the parent compound, or their cumulative mass. Administration may be intravenous. The method may include detecting the binding of the compound to the detectable target protein. In some embodiments, the method further includes utilizing light activation and the emission of a detectable signal. In some embodiments, the method includes comparing the signal to a control value, wherein an increase in the signal compared to the control value indicates the presence of a detectable target protein, wherein the control value is the signal in the absence of a detectable target protein. In some embodiments of the method, the detectable signal is a fluorescent signal or an infrared signal. In some embodiments, the light is a laser.

[0152] In some embodiments, this disclosure provides a method for detecting a detectable target protein, such as amyloid-β protein or phosphorylated tau protein, or their cumulative mass. The method includes contacting a compound described herein with a tissue or sample that may contain a detectable target protein, such as amyloid-β protein or phosphorylated tau protein, or their cumulative mass, wherein the compound binds to the detectable target protein. In some embodiments, this disclosure provides a method for detecting the presence or absence of binding of a compound described herein or its parent compound to a detectable target protein, including administering the compound or its pharmaceutically acceptable salt to a patient, as described herein. In some embodiments, this disclosure provides a method for monitoring the response of a patient suffering from a disease or symptom characterized by the presence of a detectable target protein to treatment, including binding an effective amount of the compound to the detectable target protein after treatment. A pharmaceutically acceptable salt is used herein or its salt, and a signal generated in response to binding is detected, wherein a decrease in signal compared to before treatment indicates a patient response to treatment. In some embodiments, the detectable target protein is amyloid protein or an amyloid protein such as an Aβ peptide, a viral peptide, an α-synuclein, or superoxide dismutase. In some embodiments, amyloid protein or amyloid protein is β-amyloid (1-42) (Aβ(1-42)). In some embodiments, the detectable target protein is phosphorylated tau protein. In some embodiments, phosphorylated tau protein is triple-repeated tau or quadruple-repeated tau.

[0153] In some embodiments, detection is performed within approximately 1 second, 5 seconds, 1 minute, 10 minutes, 30 minutes, or 60 minutes after contact with the compound and administration of the compound. In some embodiments, detection is performed within approximately 1–5 minutes after contact with the compound or administration of the compound.

[0154] Imaging devices facilitate in situ detection of the binding of target proteins (e.g., amyloid-β protein or phosphorylated tau protein) or substances thereof to compounds described herein, preferably handheld or portable. Retinal imaging devices may include lenses and image sensors, and optionally, a laser source. When the source emits laser light toward the retina, if Aβ accumulates on the retina and binds to an Aβ-binding probe, this accumulation can be easily detected and quantified by lenses and image sensors that collect and sense fluorescence signals. The imaging device can be any device capable of detecting light, such as a camera. The imaging device may include a confocal lens. The imaging device may be a retinal imaging device. The imaging device may include a fundus camera. The detection may include a confocal laser scanning microscope. The detection may be non-mydriatic. For an overview of retinal imaging techniques, see, for example, MDAbràmoff et al., Retinal Imaging and Image Analysis (2010), IEEE Rev Biomed Eng. 3:169–208.

[0155] Amyloid-β protein or phosphorylated tau protein may accumulate in the patient's eye. In some implementations, this contact, when activated by light, causes the emission of a detectable signal. This signal can be fluorescent or infrared.

[0156] This document provides a method for treating a patient's neurological disease or condition, comprising administering to the patient a compound described herein. The compound may be a compound of formula I. In some embodiments, the compound is compound 7, compound 8, or compound 9. In some embodiments, the compound is compound 7, compound 8, compound 9, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is compound 10.

[0157] Dosage and pharmaceutical composition

[0158] Pharmaceutical compositions of the compounds described herein for administration to patients are also provided. The compound may be a compound of formula I. In some embodiments, the compound is compound 7, compound 8, or compound 9. In some embodiments, the compound is compound 7, compound 8, compound 9, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is compound 10: the compound may be administered in single or multiple doses. The compound may be administered by various methods, including, for example, rectal, buccal, intranasal, and transdermal routes. In some embodiments, the pharmaceutical composition may be administered by intra-arterial injection, intravenous, intraperitoneal (“ip”), parenteral, intramuscular, subcutaneous, oral, topical, or as an inhaler. In some embodiments, the compound described herein is administered intravenously. Intravenous administration may be bolus injection or continuous injection. Other injection routes include: intra-arterial, intracardiac, intrasheath, intra-bone, intra-articular, intrasynovial, intradermal, subcutaneous, intramuscular and intradermal, intracranial, intralesional, and intratumoral.

[0159] In some embodiments, the compounds described herein are administered to the eye. In some embodiments, the compounds are administered topically to the eye. In some embodiments, administration is parenteral, such as by injection. In some embodiments, the compounds described herein are administered as a bolus, such as in the arm. In some cases, the compounds are administered topically to the eye. In some embodiments, administration is oral.

[0160] This compound may be effective over a wide dose range. In some embodiments, the dose is 0.01 to 1000 mg, 0.5 to 100 mg, 1 to 50 mg, or 5 to 40 mg daily. Exemplary doses include 10, 20, 30, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, 900, and 1000 mg. In some embodiments, the effective amount of the compound corresponds to about 50 to 500 mg. The effective amount can vary among patients. The exact dose will depend on the route of administration, the form of the compound, the subject to be treated, the weight of the subject to be treated, and the preferences and experience of the attending physician.

[0161] In some embodiments, the effective amount of the compound is about 0.01-1000 mg per dose. In some embodiments, the effective amount of the compound is 50-500 mg per dose. In some cases, the effective dose corresponds to approximately 0.01-100 mg, 0.01-200 mg, 0.01-300 mg, 0.01-400 mg, 0.01-500 mg, 0.01-600 mg, 0.01-700 mg, 0.01-800 mg, 0.01-900 mg, 0.01-1000 mg, 0.1-100 mg, 0.1-200 mg, 0.1-300 mg, 0.1-400 mg, 0.1-500 mg, 0.1-600 mg, 0.1-700 mg, 0.1-800 mg, 0.1-900 mg, 0.1-1000 mg, 1-100 mg, 1-200 mg, 1-300 mg, 1-400 mg, 1-500 mg, 1-600 mg per person per dose. mg, 1-700 mg, 1-800 mg, 1-900mg, 100-200 mg, 100-300 mg, 100-400 mg, 100-500 mg, 100-600 mg, 100-700 mg, 100-800mg, 100-900 mg, 100-1000 mg, 200-300 mg, 200-400 mg, 200-500 mg, 200-600 mg, 200-700mg, 200-800 mg, 200-900 mg, 200-1000 mg, 300-400 mg, 300-500 mg, 300-600 mg, 300-700mg, 300-800 mg, 300-900 mg、300-1000 mg, 400-500 mg, 400-600 mg, 400-700 mg, 400-800mg, 400-900 mg, 400-1000 mg, 500-600 mg, 500-700 mg, 500-800 mg, 500-900 mg, 500-1000 mg, 600-700 mg, 600-800 mg, 600-900 mg, 600-1000 mg, 700-800 mg, 700-900 mg, 700-1000 mg, 800-900 mg, 800-1000 mg or approximately 900-1000 mg.In some cases, the effective dose corresponds to approximately 50-100 mg, 50-400 mg, 50-500 mg, 100-200 mg, 100-300 mg, 100-400 mg, 100-500 mg, 200-300 mg, 200-400 mg, 200-500 mg, 300-400 mg, 300-500 mg, or 400-500 mg per adult dose.

[0162] In some embodiments, the compound is administered in a single dose. In some embodiments, the compound is administered in multiple doses.

[0163] In some embodiments, the compound is administered in a pharmaceutical composition comprising a liquid carrier, for example, for intravenous administration. In some embodiments, the volume of the pharmaceutical composition is from about 10 μL to about 1000 mL. For example, the volume can be about 10 µL, 50 µL, 100 µL, 300 µL, 500 µL, 1 mL, 10 mL, 50 mL, 100 mL, 200 mL, 300 mL, 400 mL, 500 mL, 600 mL, 700 mL, 800 mL, 900 mL, or 1000 mL.

[0164] In some embodiments, the compound is administered as drops. In some embodiments, the size of the administered droplets is in the range of about 10-100 μL, about 20-50 μL, or about 50-80 μL. In some embodiments, several drops are administered per dose, for example, 1-3 times per dose, 3-10 drops per dose, or 7-10 drops per dose. In one embodiment, the formulation of this disclosure is administered as about 1 drop per dose, 1-6 times daily.

[0165] In some embodiments, the compounds described herein are formulated into pharmaceutical compositions. In certain cases, pharmaceutical compositions are formulated in a conventional manner using one or more physiologically acceptable carriers, which include excipients and adjuvants that facilitate the processing of the active compound into a pharmaceutically acceptable formulation. Proper formulation depends on the chosen route of administration. Any pharmaceutically acceptable technique, carrier, and excipient suitable for formulating the pharmaceutical compositions described herein may be used: Remington: The Science and Practice of Pharmacy, 19th edition (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, HA and Lachman, L., eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th edition (Lippincott Williams & Wilkins 1999). In some embodiments, a pharmaceutical composition is provided comprising the compounds described herein and a pharmaceutically acceptable carrier.

[0166] The pharmaceutical compositions provided herein comprise the probes described herein and pharmaceutically acceptable diluents, excipients, or carriers. The compound may be a compound of Formula I. In some embodiments, the compound is compound 7, compound 8, or compound 9. In some embodiments, the compound is compound 7, 8, 9, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is compound 10: in certain circumstances, such as in combination therapy, the compound is administered as a pharmaceutical composition wherein one or more probes are mixed with other active ingredients. In some embodiments, the pharmaceutical composition comprises one or more compounds described herein.

[0167] As used herein, a pharmaceutical composition refers to a mixture of the compound described herein with other chemical components (e.g., carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and / or excipients). This pharmaceutical composition facilitates the administration of the compound to a patient. In certain cases used to practice the treatments or uses provided herein, a therapeutically effective amount of one or more probes provided herein is administered in the form of a pharmaceutical composition to two mammals suffering from a disease or symptom to be detected, diagnosed, or treated. In some embodiments, the patient is a person. In some cases, the therapeutically effective amount depends on the severity of the disease, the age and relative health status of the subject, the potency of the compound used, and other factors. The compounds described herein are used alone or in combination with one or more therapeutic agents as components of a mixture.

[0168] For administration by injection, the compounds described herein may be dispersed in a liquid pharmaceutically acceptable medium. A liquid pharmaceutically acceptable medium can be any aqueous or non-aqueous medium known in the art. Examples of aqueous media include aqueous solutions of physiological saline, solutions of sugars such as glucose or mannitol, and pharmaceutically acceptable buffer solutions. In some embodiments, the aqueous carrier is a physiologically compatible buffer, such as Hank's solution, Ringer's solution, aqueous acetate solution, aqueous citrate solution, aqueous carbonate solution, aqueous phosphate solution, aqueous succinate solution, aqueous lactate solution, or physiological saline buffer. Examples of non-aqueous carriers include: fixed vegetable oils, glycerol, polyethylene glycol, alcohols, and ethyl oleate. The carrier may further include: antimicrobial preservatives, antioxidants, tensioning agents, buffers, stabilizers, surfactants, and other components. The pharmaceutical composition may contain cyclodextrin, such as sulfobutyl ether β-cyclodextrin or hydroxypropyl β-cyclodextrin.

[0169] The pharmaceutical compositions of the compounds described herein can be used for parenteral administration, such as by injection. The compounds for injection can be prepared, for example, as an aqueous or oil suspension or emulsion in an injection medium. Injection media may include: castor oil (castor seed oil), (ethoxylated) castor oil, sesame oil, soybean oil, corn oil, cottonseed oil, or peanut oil, as well as e-agent, mannitol, dextrose, benzyl alcohol, PEG 400, ethylene glycol, polysorbate 20, diethylene glycol monoethyl ether, 10% poloxamer 188 aqueous solution, glycerin, 10% poloxamer 407 or poloxamer 124 aqueous solution.

[0170] In some embodiments, the pharmaceutical formulation comprises one or more surfactants. Surfactants are hydrophobic or amphiphilic materials (i.e., comprising both hydrophilic and hydrophobic components or regions). Surfactants can be used to modify the surface properties of particles and alter the manner in which particles are dispersed, emulsified, or suspended. In some embodiments, the surfactant comprises lipids. Lipids that can be used include the following classes: fatty acids and their derivatives, monoglycerides, diglycerides, and triglycerides, phospholipids, sphingolipids, cholesterol and steroid derivatives, terpenes, prostaglandins, and vitamins. Examples of fatty acids include: lauric acid, phytic acid, myristic acid, palmitoleic acid, fucoidan, and oleic acid, and their mono, di, and triglycerides. Such monoglycerides, diglycerides, and triglycerides include, for example: digalactosyl diglyceride, 1,2-dioleoyl-sn-glycerol, 1,2-dispalmitoyl-sn-3-succinoglycerol, and 1,3-dispalmitoyl-2-succinoglycerol. In some embodiments, the surfactant comprises phospholipids. Permissible phospholipids include phosphatidic acid, phosphatidylcholine (which contains both saturated and unsaturated lipids), phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine, phosphatidylinositol, lysophosphatidyl derivatives, cardiolipin, and β-acyl-γ-alkyl phospholipids. Permissible steroids include cholesterol, cholesterol sulfate, cholesterol hemisuccinate, 6-(5-cholesterol-3β-alkoxy)hexyl-6-amino-6-deoxy-1-thio-α-D-galactopyranoside, 6-(5-cholesterol-3β-alkoxy)hexyl-6-amino-6-deoxy-1-thio-α-D-mannopyranoside, cholesterol (4'-trimethylammonium)butyrate, and sodium deoxycholate (NaDOC). Surfactant products include Tween 20, Tween 80, and Neobee M-5.

[0171] Other surfactants include: ethoxylated sorbitol esters, sorbitol esters, fatty acid salts, sugar esters, Protonix, Tetronics, ethylene oxide, butylene oxide, propylene oxide, anionic surfactants, cationic surfactants, mono- and diacylglycerols, mono- and diacylethylene glycol, mono- and diacylethylene glycol sorbitol, mono- and diacylglycerol succinates, alkyl acylphospholipids, fatty alcohols, fatty amines and their salts, fatty ethers, fatty esters, fatty amides, fatty carbonates, cholesterol esters, and cholesterol esters. Alcohol amides and cholesterol ethers, aluminum monostearate, ammonium dodecyl sulfate, calcium stearate, calcium dioctyl sulfosuccinate, potassium dioctyl sulfosuccinate, sodium dioctyl sulfosuccinate, emulsified waxes, magnesium dodecyl sulfate, potassium oleate, sodium castor oil, sodium cetearyl sulfate, sodium dodecyl ether sulfate, sodium dodecyl sulfate, sodium dodecyl sulfoacetate, sodium oleate, sodium stearate, sodium stearate fumarate, sodium tetradecyl sulfate, zinc oleate, zinc stearate, benzalkonium chloride, cetrimetide, cetrimetide bromide, and hexadecylpyridine chloride.

[0172] Oral administration may be another route of administration for the compositions described herein. The pharmaceutical compositions may be in the form of capsules or enteric-coated tablets, for example. Therefore, the compounds described herein may be diluted with excipients and / or within a carrier. When excipients are used as diluents, they may be in the form of solid, semi-solid, or liquid materials, acting as excipients, carriers, or media for the active ingredient. Therefore, the compositions may be in the form of tablets, pills, powders, lozenges, capsules, elixirs, suspensions, emulsions, solutions, syrups, aerosols (in solid or liquid form), ointments (containing, for example, up to 10% by weight of the active compound), soft and hard gelatin capsules, sterile injectable solutions, and sterile encapsulated powders.

[0173] Some examples of suitable excipients include: lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, astragalus gum, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup, and methylcellulose. The formulation may also contain: lubricants such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifiers and suspending agents; preservatives such as methylparaben and propylparaben; sweeteners; and flavoring agents.

[0174] In some cases, the compounds described herein are formulated for ocular administration. In some cases, ocular formulations are liquids (in the form of solutions, suspensions, reconstituted powders, sol-to-gel systems), semi-solids (ointments and gels), solids (ocular formulations), and intraocular dosage forms (injections, irrigation solutions, and implants).

[0175] This document provides ophthalmic formulations containing the compounds described herein and ophthalmologically acceptable components. Ophthalmic formulations may be administered in any form suitable for ophthalmic drug administration, such as as a solution, suspension, ointment, gel, liposome dispersion, colloidal microparticle suspension, or the like, or in ophthalmic inserts, such as in an optionally biodegradable controlled-release polymer matrix.

[0176] A "pharmaceutically acceptable" or "ophthalmologically acceptable" component means a component that is not biologically or otherwise undesirable, meaning that the component can be incorporated into the ophthalmic formulation of the present invention and administered topically to the patient's eye without causing any undesirable biological effects or interacting in a harmful manner with any other components contained in the formulation composition. When the term "pharmaceuticalally acceptable" is used to refer to a component other than a pharmacologically active agent, it indicates that the component has met the requirements of toxicological and manufacturing testing, or is included in the Inactive Ingredient Guide prepared by the U.S. Food and Drug Administration.

[0177] Ophthalmic preparations may be suitable for topical administration to the eye in the form of suspensions or emulsions. Ophthalmic preparations may contain ophthalmologically acceptable carriers. Such carriers include, for example, water; mixtures of water such as phosphate buffers, boric acid, sodium chloride, and sodium borate; and water-miscible solvents such as lower alcohols, aryl alcohols, polyalkylene glycols, carboxymethyl cellulose, polyvinylpyrrolidone, and isopropyl myristate. Ophthalmic preparations may also contain one or more excipients, such as emulsifiers, preservatives, humectants, and thickeners. For example, ophthalmic formulations may contain polyethylene glycol 200, 300, 400, and 600; polyethylene glycol 1,000, 1,500, 4,000, 6,000, and 10,000; antibacterial ingredients such as quaternary ammonium compounds, phenylmercuric salts, thimerosal, methylparaben and propylparaben, benzyl alcohol, and phenethyl alcohol; buffers such as sodium borate, sodium acetate, and gluconate buffers; and other reagents such as sorbitol monolaurate, triethanolamine, oleate, polyoxyethylene sorbitol monopalmitate, sodium dioctyl sulfosuccinate, monothioglycerol, thiosorbitol, and ethylenediaminetetraacetic acid. The ophthalmic formulation may be isotonic. The ophthalmic formulation may also contain surfactants or stabilizers. Surfactants include Carbopol®. Stabilizers include sodium bisulfite, sodium metabisulfite, and sodium thiosulfate.

[0178] The formulation may contain an effective amount of a penetration enhancer that promotes the penetration of formulation components through cell membranes, tissues, and the extracellular matrix (including the cornea). An "effective amount" of penetration enhancer means a measurable increase in concentration sufficient to provide penetration of one or more formulation components through membranes, tissues, and the extracellular matrix, as just described. Suitable penetration enhancers include, for example, methanesulfonylmethane (MSM; also known as methyl sulfone), a combination of MSM and dimethyl sulfoxide (DMSO), or, in a less preferred embodiment, a combination of MSM and DMSO, wherein MSM is particularly preferred.

[0179] Kits and Packaging

[0180] The kits provided herein include the compounds described herein, imaging devices, and optional suitable packaging. The imaging device may be a retinal imaging device. In some embodiments, the kit further includes instructions for use.

[0181] The imaging apparatus may include a lens and an image sensor for detecting the emitted signal. In some embodiments, the retinal imaging apparatus detects a fluorescence signal. In some embodiments, the retinal imaging apparatus also includes a laser source that can be used to activate the fluorescence signal. The imaging apparatus may include a suitable retinal scanner.

[0182] Table of Abbreviations and Acronyms

[0183]

[0184]

[0185] Example

[0186] This document includes the following embodiments to disclose specific implementations of this disclosure. Those skilled in the art will understand that the techniques disclosed in the following embodiments represent techniques that work well in the practice of this disclosure and can therefore be considered as constituting a particular mode of implementation. However, those skilled in the art will understand that many changes can be made to the specific embodiments disclosed based on the content of this disclosure without departing from the spirit and scope of this disclosure and still obtaining the same or similar results.

[0187] Example 1

[0188] Based on preliminary rabbit pharmacokinetic studies of various formulations of the methyl ester derivative of compound 7, studies were conducted to modify and improve certain physicochemical properties. The hydroxyl compound (i.e., compound 7) was found to exhibit fluorescent properties suitable for use with conventional instruments. Importantly, compound 7 also showed the ability to detect Aβ deposits in human Alzheimer's disease brain tissue, both in vitro and ex vivo.

[0189] synthesis

[0190]

[0191] Synthesis of (6-bromonaphth-2-yl)methanol(2)

[0192] In N2 at 0 °C, methyl 6-bromo-2-naphthoate (1) (50.0 g, 189 mmol) dissolved in 500 mL anhydrous THF was added dropwise to a solution of LiAlH4 (8.2 g, 217 mmol dissolved in 500 mL THF). The reaction mixture was stirred at 0 °C for 1 hour. After the reaction was completed as monitored by TLC, the mixture was treated with H2O, 15% NaOH, H2O (1:1:3, v / v / v). After filtration, the filtrate was concentrated and extracted with EA, and dried over NaSO4. The crude product was purified from (PE:EA = 3:1) to obtain the title compound.

[0193] Synthesis of 6-bromo-2-naphthaldehyde (3)

[0194] PCC (76.4 g, 354 mmol) was added to a suspension of (6-bromonaphth-2-yl)methanol (2) (42.0 g, 177 mmol) and silica gel (76.4 g) in DCM (500 mL). The reaction mixture was stirred at RT (room temperature, 25 ± 5 °C) for 1.5 h. After completion, the mixture was filtered through a silica mat and concentrated under reduced pressure to obtain the title compound.

[0195] Synthesis of 6-(piperidin-1-yl)-2-naphthaldehyde (4)

[0196] In dried and degassed toluene (300 mL), Pd(OAc)₂ (1.5 g, 6.3 mmol), BINAP (4.4 g, 7.1 mmol), 6-bromo-2-naphthaldehyde (26) (30.0 g, 127.8 mmol), Cs₂CO₃ (60.0 g, 183.9 mmol), and piperidine (12.7 g, 149.5 mmol) were added. The reaction mixture was stirred at 115 °C for 8 hours. After cooling, the mixture was filtered, washed with EA, and then concentrated to one-third of its volume. 200 mL of 6N hydrochloric acid was added with vigorous stirring. The aqueous phase was separated, extracted three times with DCM, adjusted to alkalinity with 5N NaOH, and then extracted again with EA. The organic phase was concentrated to give the crude product, which was further purified by silica gel chromatography (PE:EA = 20:1 to 2:1) to obtain the title compound.

[0197] Synthesis of 2-cyano-N-(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)acetamide (6)

[0198] In a pear-shaped flask, 5 g (6.0 g, 40 mmol) was added to methyl 2-cyanoacetate (4.0 g, 40 mmol) with stirring. The mixture was stirred overnight at room temperature and then concentrated to a crude product. This crude product was used directly in the next step.

[0199] Synthesis of (E)-2-cyano-N-(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)-3-(6-(piperidin-1-yl)naphth-2-yl)acrylamide (compound 7)

[0200] 6-(piperidin-1-yl)-2-naphthaldehyde (4) (7.0 g, 29.3 mmol) and 2-cyano-N-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)acetamide (6) (7.9 g, 36.5 mmol) were dissolved in anhydrous THF (250 mL), and piperidine (0.5 g, 5.9 mmol) was added. The resulting mixture was refluxed for 12 hours. The reaction mixture was then concentrated under reduced pressure to a crude substance, which was purified by silica gel chromatography to give the title compound. Exact weight 437.23; m / e 437.23 (100%), 438.23 (28.9%), 439.24 (4.7%); Elemental analysis (C 25 H 31 N3O4): C 68.63%, H 7.14%, N 9.60%, O 14.63%.

[0201] Properties of Compound 7

[0202] The physical properties of compound 7 were determined. The solubility of compound 7 in various media is given in Table 1.

[0203] Table 1. Solubility of Compound 7

[0204]

[0205] Due to its poor water solubility (Table 1), a feasible nano-suspension formulation was explored. The aim was to achieve an average particle size of less than 0.5 µm. Table 2 lists the particle size data for compound 7 in the suspension formulation.

[0206] Table 2. Particle size of compound 7 in the suspension formulation (NaDOC = sodium deoxycholate)

[0207]

[0208] Example 2

[0209]

[0210] Synthesis of Compound 8

[0211] Using traditional methods (such as POCl3) )Attempts to prepare compound 9 with a base (such as triethylamine) proved unsuccessful. Due to the high reactivity of certain functional groups present in compound 7, such as the NH amide found to react negatively with POCl3, an alternative synthetic route was needed. Other bases, such as LDA and LiHMDS, yielded mixtures of products by TLC without separation. It was determined that tetrabenzyl pyrophosphate with sodium hydride provided compound 8. Compound 8 was prepared according to the following steps.

[0212] At 0°C, 1.6 g (3 mM) of tetrabenzyl pyrophosphate, prepared using the Merck organic synthesis method, was added to 1 g (2.3 mM) of compound 7 dissolved in 15 mL THF, resulting in a deep red solution. Sodium hydride (100 mg, 2.5 mM, 60% oil) was added. After 15 minutes, the temperature was raised to room temperature, and a solid began to precipitate. DMF (5 mL) was added, and the mixture was stirred at RT for 1 hour. Water (200 mL) and ethyl acetate (200 mL) were added. The ethyl acetate layer was dried and evaporated. The title compound was obtained by purification using ISCO® on an 80 g silica gel column with 0–100% hexane / ethyl acetate. Figures 1A to 1C Compound 8 is given in 1 HNMR spectrum. MS (m / z) 701.3 [M+H] + .

[0213] Synthesis of Compound 9

[0214] 1 gram (compound 8) of 95% ethanol (150 mL) degassed with argon was added. 120 mg of 10% Pd / C was added, and hydrogen was bubbled into the reaction mixture for 5 minutes. The mixture was then stirred in a hydrogen balloon for 3 hours. The reaction mixture was degassed with argon and evaporated. Purification was performed by preparative LC / MS on a 25 × 250 mm C18 column using 0–100% water (2 g of ammonium acetate per liter). After free drying, compound 9 was obtained. Figure 2A and Figure 2B Compound 9 is given in the text. 1 1H NMR spectrum (D2O).

[0215] It was discovered that the above-mentioned synthesis using hydrogenation produces byproducts via, for example, the reduction of double bonds. This reduction leads to lengthy and tedious purification by HPLC.

[0216] Synthesis of diammonium salt of compound 9 (also known as compound 10)

[0217] It was found that using trimethylsilyl bromide for a short time at zero degrees Celsius reduced side reactions and the need for chromatography.

[0218]

[0219] A solution of compound 8 (5 mmol, 3.49 g, 1 equivalent) in anhydrous CH₂Cl₂ (100 mL) was cooled to 0 °C under argon atmosphere, and trimethylsilyl bromide (50 mmol, 6.8 mL, 10 equivalent) was added using a syringe. The reaction mixture was stirred at 0 °C for 30 min, and the completion of the conversion was monitored by HPLC. The mixture was then quenched with MeOH (50 mL) and stirred for 10 min. The solution was evaporated to dryness; this step was repeated four times.

[0220] The organic solvent was evaporated under vacuum, and the residue was suspended in trace amounts of MeOH. EtOAc was then added to precipitate phosphonic acid. The residue was filtered and washed with EtOAc (×2). The residue was then dried under vacuum to obtain the desired phosphoric acid.

[0221] Phosphonic acid was treated with NH4OAc (25 mmol, 1.93 g, 5 equivalents) and 150 mL of water at room temperature and stirred for 15 min to give a clear red / orange solution. The reaction mixture was then lyophilized to give the final product. LC-MS: (ES, m / z )518 [M+1] + . 1 H-NMR: (400MHz, CD3OD) δ 8.31–8.22 (m, 2H), 8.08 (dd, J = 8.8, 1.9Hz, 1H), 7.81 (d, J = 9.2 Hz, 1H), 7.74 (d, J = 8.8 Hz, 1H), 7.40 (dd, J =9.2, 2.5 Hz, 1H), 7.18 (d, J = 2.5 Hz, 1H), 4.08–3.98 (m, 2H), 3.79–3.65 (m,8H), 3.58 (t, J = 5.5 Hz, 2H), 3.45–3.39 (m, 4H), 1.86–1.63 (m, 6H).

[0222] Solubility of compound 9

[0223] Compound 9 was found to have a solubility of 20-30 mg / mL in phosphate buffer at pH 7.4.

[0224] Pharmacokinetics of Compound 9

[0225] Compound 9 was administered to rabbits as a solution in 0.1 M phosphate buffer at pH 7.4, with rabbits receiving either 30 mg of compound 9 (Study 1) or 28 mg of compound 9 (Study 2). The aim of these studies was to demonstrate that compound 9 can be readily converted to compound 7 in vivo.

[0226] Study 1: Rabbits received 1.5 mL of a 20 mg / mL solution of compound 9 in 0.1 M phosphate buffer (pH 7.4). Rabbits were sacrificed at the specified time points in the table below (N = 3 at each time point), and the average concentrations of compounds 9 and 7 in the retina (Table 3), brain (Table 4), and blood (Table 5) were measured.

[0227] Table 3. Average retinal concentrations of compounds 9 and 7.

[0228]

[0229] Table 4. Average brain concentrations of compound 9 and compound 7.

[0230]

[0231] Table 5. Mean blood concentrations of compound 9 and compound 7.

[0232]

[0233] Study 2: The aim of this study was to determine the presence of high concentrations of Compound 9 and / or Compound 7 in specified tissues at earlier time points. Rabbits received 1.4 mL of a 20 mg / mL solution of Compound 9 dissolved in 0.1 M phosphate buffer (pH 7.4). Two minutes after administration of Compound 9, blood was collected from the rabbits (N = 6) to determine the blood concentration at this time point (Table 8). The rabbits were then sacrificed at the specified time points in the table below (N = 3 at each time point), and the mean concentrations of Compound 9 and Compound 7 in the retina (Table 6), brain (Table 7), and blood (Table 7) were measured (Table 8).

[0234] Table 6. Mean retinal concentrations of compounds 9 and 7.

[0235]

[0236] Table 7. Average brain concentrations of compound 9 and compound 7.

[0237]

[0238] Table 8. Average plasma concentrations of compound 9 and compound 7.

[0239]

[0240] Based on this data, it can be concluded that in a healthy rabbit model, compound 9 is rapidly converted into compound 7 in vivo.

[0241] Modifying the terminal alcohol group of compound 7 to a phosphate ester significantly improves its water solubility. A beneficial characteristic of the diammonium salt of compound 9 is the immediate breakdown of the phosphate group by phosphatases in the blood, thereby generating compound 7. Compound 7 has been shown to be delivered across the blood-brain barrier (BBB) ​​via administration of the diammonium salt of compound 9, and more importantly, delivered to the retinal tissue. Compound 9 is rapidly converted to compound 7 (K of compound 7). p，脑 It peaked at 5 minutes after administration and decreased significantly to an undetectable value at 30 minutes. The K of compound 9... p , 脑 <0.001 indicates that it was primarily converted to compound 7 before reaching the brain. Compound 7 had a relatively short circulation time in the retina, with probe levels decreasing significantly within one hour. Importantly, compound 7 was delivered to the retinal tissue at a high concentration (682 ng / g), while compound 9 remained undetectable. Furthermore, no adverse reactions were observed in rabbits throughout this study. Based on the data shown below, it can be concluded that compound 9 is rapidly converted to compound 7, which can (1) be detected by in vitro fluorescence of Aβ40, (2) emit fluorescence at wavelengths detectable by conventional instruments, and (3) penetrate into the retina when administered systemically as compound 9 (e.g., the diammonium salt of compound 9).

[0242] Example 3

[0243] Compound 9 was administered intravenously to mouse models with detectable target proteins (e.g., amyloid β protein or phosphorylated tau protein or their accumulated amounts). The mouse retina was removed and fixed onto a slide. The retina was washed twice with PBS over 5 minutes. 98% formic acid solution was added over 5 minutes to recover the antigen. The sample was washed twice with distilled water for 5 minutes each time. The sample was equilibrated in 1× PBS for 15 minutes, and then blocked with 10× goat / donkey serum in 1× PBST (depending on the antibody) for 1 hour. The sample was covered with foil and washed three times with 1× PBS for 5 minutes each time. The sample was stained in black with DAPI (300 nM or 100 ng / mL) for 10 minutes, and then the tissue was washed with PBS 3×10 minutes each time. Anti-fading DAKO mounting medium was added, a coverslip was placed, and the tissue was held under the foil until imaging.

[0244] Fluorescence imaging studies of the samples were performed on a Leica DMI 4000B microscope (Leica GmbH, Germany) equipped with a TCS SPE camera and Leica 10, 20, and 40X objectives. The following lasers were used to visualize the fluorescent probes associated with DAPI (blue, nuclear staining), compound 7 (green), and hyperphosphorylated tau (red): 408, 488, and 568 nm. Z-stack images were captured at 40x magnification in 0.5 µm increments to visualize the entire thickness of the tissue. Hyperphosphorylated triplet tau protein was detected in the retina, generating a detectable fluorescent signal, while immunostaining with triplet tau antibodies was not detected in age-appropriate wild-type mice. In conclusion, this study demonstrates that compound 9 can be used as a diagnostic agent for the detection of amyloid-β protein or hyperphosphorylated tau protein.

[0245] Example 4

[0246] This embodiment was performed to determine whether compound 9 was detected in the retina of a TBI mouse model.

[0247] Compound 9 was administered intravenously to shockwave-induced model mice 24 hours after injury and to uninjured mice. The mouse retinas were scanned using a scanning laser ophthalmoscopy (SLO). The resulting images showed the presence of Aβ plaques.

[0248] Alternatively, or in addition to retinal scanning, retinal tissue was removed and stained with anti-Aβ antibody (6E10). Mice with shock injuries showed an immunoreactivity to Aβ in their retinal tissue, while uninjured mice showed no reactivity to 6E10. Compound 7 fluorescently labeled the retinal deposits, which were visible upon fluorescence activation, but showed no fluorescence enhancement in the tissues of uninjured mice.

[0249] Example 5

[0250] In vivo detection of retinal Aβ aggregation

[0251] In vivo retinal imaging studies were conducted in mice before and after controlled cortical impaction (CCI). The CCI model involves a controlled impact on the intact dura mater following craniotomy and is a commonly used animal model for total brain injury (TBI).

[0252] Mice were imaged prior to TBI via CCI to obtain baseline retinal images. Anesthetized mice received intravenous administration of compound 9 diammonium salt (compound 10) (15 mg / kg in 20 mg / mL 0.1 M phosphate-buffered saline, pH 7.4) before and 24 hours after CCI. Figure 3As shown, enhanced background fluorescence was observed in the retinal vascular system 3 minutes after administration of compound 10 to uninjured mice. This enhancement was no longer observed 15 minutes after administration of compound 10. Figure 3 (Top row).

[0253] Twenty-four hours after baseline imaging, mice were anesthetized, their heads fixed in a stereotactic frame, and a craniotomy was performed on the right side of the motor cortex. Using a stereotactic impactor, mice were impacted on the right side of the motor cortex with a piston at a velocity of 5 m / s, a depth of 2 mm, and a diameter of 3 mm. After injury, the incision was closed with sutures, anesthesia was terminated, and the animals were placed on a heating pad to maintain normal core temperature. Twenty-four hours after CCI, real-time retinal imaging was performed on anesthetized mice following an intravenous injection of compound 10. Figure 3 As shown, the retinal vascular system remained illuminated during the 15-minute imaging period, indicating that Aβ can be present in blood vessels. Figure 3 (bottom row). This contrasts with pre-CCI imaging, where enhanced fluorescence in the vessel is visible 3 minutes after injection, but decreases during imaging at 15 minutes.

[0254] This embodiment demonstrates that, within 24 hours of CCI, the diammonium salt of compound 9 (compound 10) can be used to detect changes in the retinal vascular system that may be indicated by the accumulation of Aβ due to TBI.

[0255] Example 6

[0256] Early diagnosis of ocular prions

[0257] In vivo fluorescence retinal imaging experiments were conducted in live ketamine-anesthetized mice using the Phoenix Micron IV fluorescence rodent retinal imaging system (similar to a commercial human retinal camera used in fluorescein angiography). 。 Following systemic administration of compound 10, mice were examined prior to clinical manifestations of disease (inoculation rate of 50% after inoculation with mouse virus (N = 5 + N = 4 controls), and inoculation rate of 75% after inoculation with mouse prion (N = 3 + N = 4 controls)) in terminal prion disease (N = 4 + N = 4 controls). Figure 4 and Figure 5 As shown, when compound 7 was administered in the form of compound 10, it successfully marked large objects around the optic disc and vascular system, which were considered to be pre-symptom markers in mice. Figure 4 ) and mice in the late stage of the disease ( Figure 5Prion deposits were present in the retina of mice. As expected, mice with simulated infection (negative control) did not show signs of retinal amyloid deposits. This was the first time that retinal virus was identified in this mouse model during the pre-symptomatic stage, indicating that compound 7, when administered as compound 10, can label prion deposits prior to clinical diagnosis.

[0258] Example 7

[0259] Detection of retinal Aβ in the diagnosis of Alzheimer's disease

[0260] In vivo analysis using human Alzheimer's disease tissue: Retinal tissue and brain from autopsies of individuals diagnosed with or possibly suffering from Alzheimer's disease were obtained (University of Florida, Miami). This study was conducted to determine: (1) whether there is a correlation between the brain of Alzheimer's disease patients and retinal amyloidosis, and (2) whether the compounds disclosed herein can be used as a diagnostic method for Alzheimer's disease by retinal fluorescence examination to identify patients with amyloid accumulation in the brain.

[0261] Colocalization experiments were performed on brain tissue from a patient diagnosed with familial Alzheimer's disease. Free-floating sections from the hippocampus of this patient after death were stained with an Aβ sequence-specific antibody (6E10) and compound 7. Figure 6 Fluorescence microscopy images in A show that the target bound by compound 7 is consistent in size and morphology with the dense core plaque of Aβ. Significantly, the binding of compound 7 does indeed exhibit immunoreactive colocalization with the dense core amyloid plaque of 6E10. Figure 6 C (white arrow), but diffuse reflective objects not stained by 6E10. One limitation of 6E10 is that it recognizes specific amino acid sequences rather than clusters of Aβ, which explains... Figure 6 B and Figure 6 Fluorescence observed in C for dense core spots and other diffuse reflective objects.

[0262] These results support the binding of compound 7 to Aβ plaques associated with Alzheimer's disease. Importantly, this demonstrates the ability of compound 7 to bind to Aβ plaques in the brain and retinal tissues of human Alzheimer's patients. This data suggests that compound 7 can bind to Aβ in human tissues, therefore it is conceivable that compound 9 or its salt (compound 10) could be used for in vivo administration of compound 7 for the diagnosis of Alzheimer's disease.

[0263] Example 8

[0264] Cerebral amyloid angiopathy (CAA)

[0265] In vivo detection of Aβ40 after systemic administration

[0266] Two different Tg mouse models were developed to induce vascular amyloid deposition in the retina associated with CAA. The accumulation of amyloid deposition in the vascular system of the brain was used to assess the impact of APP (anti-angiogenic atrophy). SwDI The PSAPP was selected, but no studies on the retina have been conducted to date. SwDI Mice expressed human APP gene (isotype 770) with mutations in human APP gene (K670N / M671L), Dutch (E693Q), and Iowsa (D694N) under the control of the mouse Thy1 promoter. PSAPP mice expressed chimeric mouse / human amyloid precursor protein (Mo / HuAPP695swe) and mutant human presenilin 1 (PS1-dE9), both directed to CNS neurons by mouse prion protein promoters. Mouse colonies were propagated and screened over time to observe the development of vascular amyloid deposits in the retina. Initially, males and females aged 12 to 16 months were examined. Within this age range, mice began to develop cataracts, making it impossible to... In the body For retinal imaging, younger mice were selected for examination with compound 10.

[0267] Retinal images are from 8-10 month old APP SwDI Acquired from (N = 6, 1 male, 5 females), PSAPP (N = 2, females), and wild-type control mice (N = 3 females). Imaging of mice was performed 5 minutes after intravenous (iv) injection of 25 μL of compound 10 (20 mg / mL in 0.1 M phosphate-buffered saline, pH 7.4). A 10-month-old female APP... SwDI Mice mainly showed small, bright fluorescent deposits along the vascular system. Figure 7 (White arrow). Fluorescence diminished over a 15-minute period, at which point the mice were euthanized and brain and retinal tissues were collected for immunohistochemistry (IHC). Figure 7 As shown, these deposits were fluorescently labeled with compound 7 in the retina and significantly co-localized with a sequence-specific antibody against Aβ (6E10). Interestingly, it demonstrates that compound 7, when administered in the form of compound 10, labeled this APP. SwDI Both blood vessels and diffuse deposits in the retinal tissue of mice ( Figure 7 (top right corner)

[0268] To better understand the brain / eye correlation, the IHC performed a brain examination. (APP) SwDI Mice showed intravascular amyloid deposition and diffuse amyloid plaques in the brain. Figure 8(White arrow), as observed in retinal IHC, indicates a good correlation. For injected PSAPP mouse compound 10 In the body Numerous deposits were observed in the lower right quadrant of the retina. Figure 7 The same deposit was identified as Aβ, as evidenced by the co-localization of compounds 7 and 6E10. In the brain, dense core plaques resembling those of Alzheimer's disease were observed, but no angioamyloid deposition was observed in the mouse brain. Figure 8 Again, this demonstrates a good brain / eye correlation, as no vascular pathology was observed in the retina of these PSAPP mice. Finally, no deposits were observed in mice injected with wild-type compounds. Compound 10 was as expected ( Figure 7 (bottom row).

[0269] In vivo analysis of human brain tissue from autopsy CAA patients

[0270] Post-mortem brain tissue from patients diagnosed with CAA was examined to mitigate the risk of conversion from a limited, robust preclinical animal model to humans. This study aimed to understand the fundamental basis for using fluorescence detection of amyloid in human tissue. Figure 9 This demonstrates that the compounds disclosed in this paper can detect amyloid protein deposits in the brains of human CAA patients.

[0271] Human CAA brain tissue and healthy control brain tissue were obtained (University of California, San Diego). Table 9 summarizes the patient population examined using Compound 7, including age, sex, and post-mortem diagnosis of the human brain tissue. All CAA patients were female, aged 70–93 years. Control brains were obtained from cognitively normal patients aged 82–94 years from both male and female donors. Free-floating portions of the midfrontal cortex from human CAA and control brains were stained with an Aβ sequence-specific antibody (6E10) and Compound 7.

[0272] Table 9. Patient population using compound 7 for ex vivo examination.

[0273]

[0274] 1 Progressive supranuclear paralysis.

[0275] Figure 9 Fluorescence microscopy images showed that compound 7 bound to the target in a consistent pattern associated with CAA amyloid pathology. Importantly, compound 7 did indeed co-localize with the immunoreactive 6E10 in CAA-filled plaques within blood vessels. Figure 9However, it was not the 6E10-stained material that diffused. One limitation of 6E10 is that it recognizes specific amino acid sequences, rather than Aβ deposits, which explains the immunoreactivity of plaques and other diffuse materials observed in CAA brains and healthy controls. Importantly, this phenomenon was observed only in CAA brains with compound 7, and not in healthy control brains. Figure 9 The results showed that compound 7 successfully stained amyloid deposits associated with CAA in human tissues.

[0276] This embodiment demonstrates that the disclosed compound was tested in vivo for Aβ40 in tg animals (administered via intravenous bolus injection). Figure 7 And Aβ40 was detected in vitro using human CAA disease tissue. Figure 9 Compound 10 was used to perform pre-death imaging of vascular amyloid deposits in the retinas of diseased mice. Ex vivo analysis of the same diseased retinas confirmed that the deposits were indeed an in vivo finding of Aβ. Therefore, the disclosed compound could provide an effective and affordable imaging agent for the diagnosis of CAA.

[0277] Example 9

[0278] Non-invasive ophthalmic diagnosis of Parkinson's disease

[0279] The following examples demonstrate that the compounds described herein provide a diagnostic test for Parkinson's disease, which may have two main implications. First, it is conceivable that currently disclosed diagnostic compounds could potentially identify patients with Parkinson's disease several years earlier than current standards, thereby allowing for earlier intervention using current therapies. Second, currently disclosed diagnostic compounds could aid in the development of improved therapies for the disease by optimizing the selection of treatment subjects, thereby increasing the likelihood of successful clinical outcomes for Parkinson's disease drug candidates.

[0280] Based on the premise of α-syn accumulation in the retina, the following model was developed and used to test currently disclosed diagnostic compounds that can target α-syn in the eye for diagnosis and monitoring of Parkinson's disease progression.

[0281] Retinal α-syn deposits were examined in two transgenic mouse models of Parkinson's disease (D and 61 strains). The selected models develop α-syn deposits in the brain, but formation in the retina has not been determined to date. Retinal images of mice were obtained before and after intravenous administration of compound 10. Following real-time imaging, mice were sacrificed, and the brain and retina were harvested for immunofluorescence and biochemical analysis.

[0282] D-Line Parkinson's Disease Tg Mouse Model:

[0283] Preliminary SYN1 staining of the ex vivo retina did not detect α-syn in the retinal tissue. As expected, compound 10 did not yield any retinal deposits in either in vivo or ex vivo settings. Therefore, the D-Line model is not a feasible model for exploring retinal α-syn deposits.

[0284] Parkinson's disease Tg mouse model No. 61:

[0285] Following systemic administration of compound 10, in 4 / 14 mice in vivo Retinal α-syn deposits were observed. Ex vivo retinal staining with the antibody SYN1 confirmed the presence of α-syn in the retinal tissue of mice from which in vivo deposition was observed. Therefore, the Line 61 model exhibits retinal α-syn in ~30% of mice and can be used in the elucidation of the utility of the disclosed compounds. As expected, WT mice (N = 10) were used as controls for evaluation, and retinal α-syn was not detected in either in vivo or ex vivo. Following final imaging, brain tissue was extracted from mouse necropsy and stained with DAPI and the SYN1 antibody to confirm the presence of α-syn in the brains of Line 61 mice.

[0286] Using the Line 61 model, it was confirmed that compound 7, administered via compound 10, is specific for α-synuclein deposits associated with Parkinson's disease and can be detected in vivo for retinal α-synuclein deposits. Figure 10 ).

[0287] To confirm that the object detected in vivo was α-syn, retinal tissue was extracted from the mouse necropsy after final imaging and stained with DAPI and SYN1 antibodies. Figure 11 ).

[0288] This embodiment demonstrates that intravenous administration of compound 10 can be used for in vivo detection of α-synuclein in the retina of mouse 61, and that fluorescence enhancement is specific for α-synuclein. Therefore, compound 7 and the anti-α-synuclein antibody (SYN1) were co-localized in the brain and retinal tissues of Tg mice.

[0289] To broaden the understanding and applicability of compound 10 as a diagnostic tool for retinal diseases in Parkinson's disease, the following studies were conducted.

[0290] To confirm that administration of compound 10 could reproducibly detect retinal α-syn deposits, Tg mice (size 61) were administered compound 10 intravenously twice, 7 days apart. Similar amounts of deposits were observed during both imaging sessions. Figure 12This indicates that administration of compound 10 can reproducibly detect retinal α-syn deposits in the same animal.

[0291] To determine the lowest dose of compound 10 that could detect retinal α-synuclein, Line 61 Tg mice were injected with compound 10 at 3 mg / kg and 15 mg / kg, with an interval of up to 6 days. The same amount of deposits was observed at both dose concentrations. Figure 13 This indicates that administration of compound 10 can detect retinal α-syn deposition at a low dose of 3 mg / kg.

[0292] In vivo staining of human Parkinson's disease eye

[0293] To understand whether compound 7 (the active drug) could be detected in the human eye for retinal α-syn, human retinal tissue was extracted from autopsies of healthy patients and patients diagnosed with Parkinson's disease, and then stained with compounds 7, DAPI, and SYN1 antibodies.

[0294] Eye Anatomy: Remove the cornea, iris, and lens from the eye patch and place the eye in 4% PFA for 1 hour. Use the macula as a guide to dissect the eye patch to estimate the direction of the eye, then restore the retina and divide the lobes into four sections (superior, inferior, temporal, and nasal). Mount the different areas on separate slides.

[0295] Planar retinal staining: Tissue was fixed in 4% PFA for 20 minutes at room temperature, washed, permeabilized, blocked (with 2.5% goat serum dissolved in PBST), stained with Alpha-synuclein antibody (SYN-1) at a 1:1000 dilution, and incubated overnight at 4°C. The tissue was washed with PBST (3 times, 5 minutes each time) and incubated in the dark at room temperature with a secondary antibody (Alexafluor 647 anti-mouse antibody) for 1 hour. The tissue was then washed with PBST (3 times, 5 minutes each time), incubated in the dark with 60 μM Compound 7 for 30 minutes at room temperature, washed with PBS (3 times, 10 minutes each time), incubated in the dark with DAPI (300 nM or 100 ng / mL) for 10 minutes, and washed with PBS (3 times, 5 minutes total). The tissue was then fixed using an antifluorescence quenching mounting medium.

[0296] As a control, a pair of retinas were extracted from a 72-year-old male who died of leukemia, brain cancer without neurological disease, or eye-related disease. The retinas were stained with compound 7 and SYN-1 Ab (α-synuclein). In autopsy tissue of the retina of healthy patients, there was little or no colocalization of compound 7 with α-synuclein (SYN-1 Ab). Figure 14 and Figure 15Exemplary images from different regions of the left and right retina are shown.

[0297] A pair of retinas were extracted from a 71-year-old male diagnosed with Parkinson's disease (PD). The retinas were stained with compound 7 and SYN-1 Ab (α-synuclein). In the retinas of human PD patients, co-localization of compound 7 and α-synuclein deposits (SYN-1 Ab) was observed in all regions. Figure 16 and Figure 17 Exemplary images from different regions of the left and right retina are shown.

[0298] To determine the number of objects co-stained with compound 7 and SYN1, the average number of objects co-stained with compound 7 and SYN1 was measured. This was done for each quadrant of both the left and right eyes (Tables 10 and 11).

[0299] Table 10. Artificial quantification of objects observed in various retinal regions of the right eye.

[0300]

[0301] Note: Each region corresponds to ~250,000 µm 2 (i.e., a 500 µm × 500 µm field)

[0302] Table 11. Artificial quantification of subjects observed in various retinal regions of the left eye.

[0303]

[0304] Note: Each region corresponds to ~250,000 µm 2 (i.e., a 500 µm × 500 µm field)

[0305] In flat retinal samples co-stained with compound 7 and SYN-1 Ab, the colocalized objects can be visualized using fluorescence. The colocalization of compound 7 and SYN-1 Ab is most prevalent at approximately two-thirds of the distance from the optic nerve and retinal margin, near the ora serrata. Colocalization of compound 7 and SYN-1 Ab is widespread in all regions, with the majority of colocalization observed in the superior region of the left retina.

[0306] Figure 18 The data indicate that compound 7 co-localizes with SYN1 in all four quadrants of a pair of Parkinson's disease eyes. These data suggest that compound 7 can be detected in human retinal tissue.

[0307] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0308] The disclosures exemplarily described herein may be suitably implemented in the absence of any one or more elements or limitations not specifically disclosed herein. Therefore, terms such as “comprising,” “including,” and “containing” should be read broadly and without limitation. Furthermore, the terms and expressions used herein have been used as descriptive terms but are not limiting, and the use of such terms and expressions is not intended to exclude any equivalent forms of the features shown and described or portions thereof, but it should be recognized that various modifications are possible within the scope of the claimed disclosure.

[0309] All publications, patent applications, patents, and other references mentioned herein are expressly incorporated by reference in their entirety, as if each reference were individually incorporated. In the event of any conflict, this specification (including definitions) shall prevail.

[0310] It should be understood that although this disclosure has been described in conjunction with the above embodiments, the foregoing description and examples are illustrative and not intended to limit the scope of this disclosure. Other aspects, advantages, and modifications within the scope of this disclosure will be apparent to those skilled in the art.

Claims

1. A compound or a pharmaceutically acceptable salt thereof.

2. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the salt is a mono salt.

3. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the salt is a di salt.

4. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the salt is selected from a sodium, potassium, lithium, ammonium, calcium, magnesium, or zinc salt.

5. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the salt is a sodium salt.

6. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the salt is a potassium salt.

7. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the salt is an ammonium salt.

8. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the salt is a diammonium salt.

9. A compound or a pharmaceutically acceptable salt thereof.

10. A pharmaceutical composition comprising a compound or pharmaceutically acceptable salt thereof according to any one of the preceding claims, and a pharmaceutically acceptable carrier.

11. Use of a compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 10 in the manufacture of a diagnostic product for determining whether a patient has a disease or condition, wherein the disease or condition is Alzheimer’s disease, cerebral amyloid angiopathy, traumatic brain injury, or Parkinson’s disease.

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