Compositions and methods for detecting traumatic brain injury

Abstract

The present disclosure relates generally to compositions and methods for determining whether a patient has traumatic brain injury (TBI) by detecting the presence of amyloid beta protein in the eye of the patient. Compositions and methods for preparing a patient to diagnose and treat traumatic brain injury (TBI) are also provided.

Description

[0001] This application is a divisional application of a patent application with application number 201980044404.1, application date May 31, 2019, and name “Compositions and methods for detecting traumatic brain injury”.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Application Serial No. 62 / 678,900, filed May 31, 2018, and U.S. Application Serial No. 62 / 733,025, filed September 18, 2018, under 35 U.S.C. §119(e), the contents of each of which are hereby incorporated by reference in their entirety into this application. Background Art

[0004] Traumatic brain injury (TBI) is a chronic condition defined as damage to the brain caused by an external force, such as a bump, blow, jolt, rapid acceleration or deceleration, or projectile penetration. Injuries resulting in TBI can result in a state of diminished or altered consciousness, leading to temporary or permanent impairment of cognitive, sensorimotor, and psychosocial functioning. Recent studies have shown that nearly 80% of military personnel who experienced blast-related, non-blast-related, mild to severe TBI suffered moderate to severe total disability within one year of the injury. In addition, several studies have shown that mild TBI is associated with post-traumatic stress disorder (PTSD), depression, and other mental and physical health problems three months after soldiers return home. In addition to these life-altering symptoms, recent studies have shown that even after a single event, TBI is considered a major risk factor for the later development of age-related neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), and chronic traumatic encephalopathy (CTE). Recent studies on veterans have shown that the overall risk of PD increases by 71% after any TBI, with even mild TBI potentially increasing the risk by 56%. Veterans were also more likely to develop PD at least two years earlier compared to veterans with PD without TBI.

[0005] The illness can be classified as mild, moderate or severe, but determining mild versus severe TBI has been problematic because physicians often refer only to existing symptoms and subjective measures to assess severity. The diagnosis of mild TBI (the majority of all brain injuries) is often missed by doctors because the symptoms present are similar to those of mental health disorders, such as bipolar disorder or depression. As a result, mild TBI is underreported and often goes untreated. The Concussion Legacy Foundation estimates that only one in six mild TBIs (also known as concussions) are diagnosed.

[0006] The diagnosis is further complicated by inconsistent diagnostic criteria, which rely heavily on patients' self-reported symptoms rather than objective examinations. The only way to definitively diagnose TBI is through a post-mortem examination of the brain using clinical evaluation and standard neuroimaging techniques.

[0007] However, it has generally been established that plaques of the misfolded protein amyloid-β (Aβ) are found within hours or days after a single TBI, similar to the hallmark Aβ plaque pathology of AD. Studies in humans and pigs shortly after TBI have shown that Aβ accumulates in damaged axons for a long time, as well as long-term accumulation of other proteins involved in the production of Aβ peptides. Recent studies have found that Aβ aggregation in the brain is as high as one-third of patients who died shortly after TBI, as well as those who survived for a year or more. In addition, about one-third of US troops injured by bomb blasts showed direct evidence of nerve fiber damage in the brain, also known as diffuse axonal injury (DAI). Even mild TBI (mTBI) can have serious consequences, inducing DAI and causing physical damage and dysfunction of injured axons. This is accompanied by repeated mild TBI (rmTBI) involving multiple mild TBIs, which may predispose injured people to exacerbated reactions and induce Aβ and tau pathology. This evidence suggests that Aβ may be an acute biomarker for diagnosing TBI.

[0008] The current standard diagnosis for severe TBI, or TBI that affects young children or adults over 60, requires invasive and expensive procedures such as intracranial pressure monitoring (ICP) and MRIs. Diagnosis of mild TBI is even more complicated, as studies have shown that less than 10% of patients with mild head injuries have positive findings on computed tomography (CT). In addition, most TBI diagnostic procedures are self-report tests, which are subjective and can be easily manipulated. Self-report tests are particularly problematic because some soldiers may be reluctant to be diagnosed and manipulate test results to avoid the stigma of being injured or separated from their squad. In addition, the Pentagon found that 60% of soldiers with symptoms of TBI refused help because they feared they would be treated differently or that their condition would prevent them from working as police officers and firefighters after retirement.

[0009] More recently, a 2018 study reported that recovery times after mild TBI were significantly longer in women than in men. Women who did not use hormonal contraceptives (HC) also had higher symptom severity relative to those who did, with this difference related to lower subjective assessments of symptom severity in women who used HC. As female military personnel comprise a large portion of the force and join during their peak childbearing years, the effects of TBI are more detrimental. There remains a significant clinical need for objective diagnosis of TBI. Summary of the invention

[0010] In some embodiments, the present disclosure provides compositions and methods capable of diagnosing traumatic brain injury (TBI) to reduce the risk of more permanent brain damage. Such methods can be rapid and non-invasive. In one example, a non-invasive fluorescent diagnostic probe capable of detecting misfolded protein amyloid-β (Aβ) can be used in a simple ophthalmic examination to detect the accumulation of Aβ in the patient's retina. The inventors have found that this detection, which can be performed by retinal imaging, can be used to quickly and reliably diagnose TBI.

[0011] According to another embodiment of the present disclosure, a rapid detection TBI kit is provided, which may include a fluorescent diagnostic probe and a portable retinal imaging device. The portable retinal imaging device can be used with an indirect ophthalmoscope or a smartphone to capture real-time retinal images at the point of care.

[0012] In one embodiment, the present disclosure provides a method for determining whether a patient has suffered a traumatic brain injury (TBI). The method may include detecting the presence of amyloid beta protein in the patient's eye. In some embodiments, the detection is for amyloid beta protein in the retina of the eye.

[0013] In some embodiments, the patient received a physical impact to the head within 30 days, 25 days, 20 days, 15 days, 10 days, 5 days, or 2 days prior to the test. In some embodiments, the physical impact was more than 1 day, 2 days, 3 days, 4 days, 5 days, 10 days, 15 days, 20 days, 25 days, or 30 days prior to the test. In some embodiments, the patient received a physical impact to the head within 24 hours prior to the test.

[0014] In some embodiments, the patient has not suffered direct physical or optical impact on the eye. In some embodiments, the patient is not known or suspected of having Alzheimer's disease. In some embodiments, the patient is a person under 40 years old.

[0015] In some embodiments, the detection comprises contacting the amyloid beta protein with the probe in vivo. In some embodiments, the contacting causes emission of a detectable signal when activated by light. In some embodiments, the detectable signal is an infrared signal. In some embodiments, the detectable signal is a fluorescent signal.

[0016] In some embodiments, the detection comprises imaging the amyloid beta protein with an ex vivo probe. In some embodiments, contacting the amyloid beta protein with the probe is performed in vivo, and the detection and / or imaging is performed ex vivo. In some embodiments, prior to the detection and / or imaging, a sample comprising the amyloid beta protein is removed from the subject.

[0017] In some embodiments, the probe comprises a compound of Formula Ic:

[0018]

[0019] in

[0020] EDG is:

[0021] a) a heterocycloalkyl group of not more than 10 carbon atoms, which is optionally substituted by one or more R 17 replace; or

[0022] b)-NR 10 R 11 ;

[0023] Each R 17 are independently halogen, -OR 18 、-NR 19 R 20 、C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons;

[0024] R 10 , R 11 , R 18 , R 19 and R 20 Each of which is independently hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbon atoms, heterocycloalkyl of up to 10 carbon atoms, arylene of up to 10 carbon atoms, or heteroarylene of up to 10 carbon atoms, each of which is optionally replaced by one or more R 21 replace;

[0025] R 21 Each of which is independently halogen, -OR 22 、-NR 23 R 24 、C1-C 10 Alkyl, C1-C 10 is a heteroalkyl, a cycloalkyl of up to 10 carbon atoms, a heterocycloalkyl of up to 10 carbon atoms, an arylene of up to 10 carbon atoms, or a heteroarylene of up to 10 carbon atoms, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally replaced by one or more R 25 replace;

[0026] R 22 , R 23 and R 24 Each of 10 Alkyl; and

[0027] Each R 25 Independently C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons;

[0028] Ar is an arylene group having not more than 14 carbon atoms or a heteroarylene group having not more than 14 carbon atoms,

[0029] each optionally substituted with one or more R1;

[0030] Each of R1 is independently halogen, -OR2, -NR3R4, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted with one or more R5;

[0031] R2, R3 and R4 are independently hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons, each of which is optionally substituted with one or more R5, excluding hydrogen;

[0032] Each R5 is independently halogen, -OR6, -NR7R8, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons;

[0033] R6, R7, R8 and R 84 are independently hydrogen or C1-C 10 alkyl;

[0034] EWG is selected from -F, -Cl, -Br, -CH=O, NO2, -CF3, -CCl3, -SO3H and -CN;

[0035] WSG is:

[0036] i)

[0037]

[0038] ii) polyethylene glycol, polypropylene glycol, copolymers of polyethylene glycol and polypropylene glycol or alkoxy derivatives thereof;

[0039] iii)

[0040]

[0041] Where n is an integer from 1 to 50 and R 81 is hydrogen, C1-C 10 Alkyl, C1-C 10 Alkenyl or C1-C 10 Alkynyl, wherein the alkyl, alkenyl or alkynyl group is optionally substituted with one or more C1-C 10 Alkyl, C1-C 10 substituted with heteroalkyl, cycloalkyl of up to 10 carbon atoms, heterocycloalkyl of up to 10 carbon atoms, arylene of up to 10 carbon atoms, or heteroarylene of up to 10 carbon atoms;

[0042] iv)

[0043]

[0044] v)

[0045]

[0046] vi)

[0047] -(C1-C 10 Alkyl)-R 33 -R 37 ,in:

[0048] R 33 is a heteroarylene group having no more than 10 carbon atoms; and

[0049] R 37 is -(C1-C6 alkyl)(heterocycloalkyl of not more than 10 carbons);

[0050] vii)

[0051]

[0052] viii)

[0053] –(C1-C 10 Heteroalkyl)-R 33 -R 37 ,in:

[0054] R 33 is a heteroarylene group having no more than 10 carbon atoms; and

[0055] R 37is -(C1-C6 alkyl)(heterocycloalkyl of not more than 10 carbons); or

[0056] ix)

[0057]

[0058] X is C=O or SO2, or X and R 84 Combine to form pyridyl;

[0059] Y is NH or S.

[0060] In some embodiments, the probe comprises an antibody. In some embodiments, the antibody is specific for the protein amyloid beta (Aβ). In some embodiments, the antibody selectively binds to misfolded protein amyloid-β (Aβ). The antibody can be labeled with a detectable marker, or can be detected by binding to a molecule associated with a detectable marker.

[0061] In some embodiments, the method further comprises determining that the patient suffers from TBI if amyloid beta is detected in the eye. In some embodiments, the method further comprises instructing the patient to avoid active physical activity. In some embodiments, the method further comprises administering to the patient an agent that treats or ameliorates TBI.

[0062] In one embodiment, a method of preparing a patient for diagnosis of traumatic brain injury (TBI) is also provided, comprising administering to the patient's eye a probe that specifically binds to amyloid beta protein. In some embodiments, the method further comprises detecting binding of the probe to amyloid beta protein in the eye. Examples of probes are described above.

[0063] In some embodiments, a method for diagnosing traumatic brain injury (TBI) is provided, comprising administering a probe that specifically binds to amyloid beta protein to the patient's eye, and detecting the binding of the probe to amyloid beta protein in vitro. In some embodiments, contacting the amyloid beta protein with the probe is performed in vivo, and the detection is performed in vitro. In some embodiments, prior to detection, a sample comprising amyloid beta protein and the probe is removed from the subject.

[0064] In some embodiments, the administration is intravenous administration or is located in the retina of the eye. In some embodiments, the patient suffered a physical impact to the head within 30 days, 25 days, 20 days, 15 days, 10 days, 5 days, or 2 days prior to the test. In some embodiments, the physical impact is more than 1 day, 2 days, 3 days, 4 days, 5 days, 10 days, 15 days, 20 days, 25 days, or 30 days prior to the test. In some embodiments, the patient suffered a physical impact to the head within 24 hours prior to the test.

[0065] In some embodiments, the patient has not suffered direct physical or optical impact on the eye. In some embodiments, the patient is not known or suspected of having Alzheimer's disease. In some embodiments, the patient is a person under 40 years old.

[0066] Also provided are kits and packages comprising a probe that specifically binds to amyloid beta protein and a retinal imaging device. In some embodiments, the retinal imaging device comprises a laser light source. In some embodiments, the retinal imaging device further comprises a retinal scanner. In some embodiments, the kit or package further comprises an ophthalmoscope. BRIEF DESCRIPTION OF THE DRAWINGS

[0067] Figure 1 Ex vivo fluorescent images of the surface of a flat-mounted human retina (superior temporal region) stained with DAPI (4',6-diamidino-2-phenylindole) and Compound 1 are presented. A) AD patient with Braak stage V / VI. B) AD patient with Braak stage V / VI. C) Cognitively normal patient. D) Cognitively normal patient. Staining is as follows: blue = DAPI nuclear staining, red = autofluorescence, green / yellow = Compound 1 staining. Scale bar = 100 μm.

[0068] Figure 2 The images presented in the upper row show that Compound 1 causes amyloid beta to fluoresce in retinal tissue of the TBI blast mouse model. Lower row: no fluorescence is visible in normal mice. Staining is as follows: green = Compound 1 staining, red = staining with 6E10 antibody, an amyloid beta specific antibody.

[0069] Figure 3 The images presented in the upper row show that Compound 1 causes amyloid beta to fluoresce in brain tissue of the TBI blast mouse model. Lower row: no fluorescence is visible in normal mice. Staining is as follows: green = Compound 1 staining, red = staining with 6E10 antibody, an amyloid beta specific antibody.

[0070] Figure 4 The images presented in the upper row show images of retinal tissue from C5BL mice subjected to controlled cortical impact (CCI), stained with: A) DAPI B) Compound 1 C) 6E10 antibody D) Merged image of the three stains. Lower row: E) Retinal tissue from C5BL / 6 mice subjected to craniotomy, stained with DAPI, Compound 1, and 6E10 antibody. F) DAPI G) Compound 1 H) 6E10 antibody I) Merged fluorescence spectra are shown on the left. Arrows indicate visible aggregated proteins.

[0071] Figure 5The images presented in the upper row show images of retinal tissue of C5BL / 6 mice subjected to CCI, stained with: A) DAPI B) Compound 1 C) 6E10 antibody D) Merged image of the three stainings. Lower row: D) Merged image of the three stainings E) Fluorescence spectrum of DAPI F) Fluorescence spectrum of Compound 1 G) Fluorescence spectrum of 6E10 antibody H) Merged fluorescence spectrum of all three stainings.

[0072] Figure 6 The images presented in the upper row show images of retinal tissue from C5BL / 6 mice subjected to CCI, stained with: A) DAPI B) Compound 1 C) 6E10 antibody D) Merged image of the three stainings. Lower row: Retinal tissue from C5BL / 6 mice subjected to craniotomy, stained with: E) DAPI F) Compound 1 G) 6E10 antibody H) Merged image of the three stainings. Arrows indicate visible aggregated proteins.

[0073] Figure 7 Presented are A) fluorescence spectrum of DAPI B) fluorescence spectrum of Compound 1 C) fluorescence spectrum of 6E10 antibody D) merged fluorescence spectrum of all three stains E) merged image showing retinal tissue of C5BL / 6 mice subjected to CCI and stained with DAPI, Compound 1 and 6E10 antibody.

[0074] Figure 8 Images of retinal staining of mice subjected to controlled cortical impact (CCI) (upper row) or Sham mice (lower row) are presented. Upper row: flat-mounted images of CCI mouse retinas stained with Compound 1, 6E10, and merged images. Arrows indicate areas of colocalization with Compound 1 and 6E10. Lower row: Sham mice stained with Compound 1, 6E10, and merged images.

[0075] Fig. 9 Images of real-time retinal imaging performed with mice using compound 23 before and after CCI are presented. Top row: Time course retinal imaging of 3-month-old pre-CCI mice after intravenous administration of compound 23. The top labeled time course represents the in vivo retinal imaging time points before (t=0) and after intravenous administration of compound 23. Bottom row: Time course retinal imaging of the same mouse using compound 23 24 hours after CCI.

[0076] It will be appreciated that some or all of the drawings are schematic for illustrative purposes. DETAILED DESCRIPTION

[0077] definition

[0078] The following description sets forth exemplary embodiments of the present technology. However, it should be appreciated that such description is not intended to limit the scope of the present disclosure, but is provided as a description of exemplary embodiments.

[0079] As used in this specification, the following words, phrases and symbols are generally intended to have the meanings set forth below, unless otherwise indicated in the context in which they are used.

[0080] The term "alkyl" by itself or as part of another substituent means a straight chain (i.e., unbranched) or branched chain, or combinations thereof, which may be fully saturated, monounsaturated, or polyunsaturated, and may include divalent and polyvalent groups, having the specified number of carbon atoms (i.e., C1-C 10 represents 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, e.g., n-pentyl, n-hexyl, n-heptyl, n-octyl, etc.

[0081] Unless otherwise stated, the term "heteroalkyl" by itself or in combination with another term refers to a stable straight or branched chain or cyclic hydrocarbon radical, or combinations thereof, consisting of at least one carbon atom and at least one heteroatom selected from O, N, P, Si and S, and wherein the nitrogen and sulfur atoms may be optionally oxidized and the nitrogen heteroatom may be optionally quaternized. The heteroatom(s) O, N, P and S and Si may be placed at any interior position of the heteroalkyl radical or at the position at which the alkyl radical is attached to the rest of the molecule. Examples include, but are not limited to, -CH2-CH2-O-CH3, -CH2-CH2-O-CH2-CH2-O-CH3, -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, -O-CH2-CH3, and -CN. Two or more heteroatoms may also be consecutive.

[0082] Unless otherwise indicated, the terms "cycloalkyl" and "heterocycloalkyl" by themselves or in combination with other terms represent cyclic forms of "alkyl" and "heteroalkyl", respectively. In addition, for heterocycloalkyl, heteroatoms may occupy the position where the heterocycle is connected to the rest of the molecule. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, etc. Examples of heterocycloalkyl include, but are not limited to, tetrahydropyran, 1-(1,2,5,6-tetrahydropyridyl), 1-piperidyl, 2-piperidyl, 3-piperidyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothiophene-2-yl, tetrahydrothiophene-3-yl, 1-piperazinyl, 2-piperazinyl, etc. Examples of heterocycloalkyl include, but are not limited to, glucose, mannose, allose, altrose, gulose, idose, galactose, and talose. Examples of heterocycloalkyl groups include, but are not limited to:

[0083] wait.

[0085] Unless otherwise indicated, the term "halo" or "halogen" by itself or as part of another substituent represents a fluorine, chlorine, bromine or iodine atom. In addition, terms such as "haloalkyl" are intended to include monohaloalkyl and polyhaloalkyl. For example, the term "halo (C1-C4) alkyl" is intended to include, but is not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl and the like.

[0086] Unless otherwise indicated, the term "aryl" refers to a polyunsaturated aromatic hydrocarbon substituent, which may be a single ring or multiple rings (preferably 1-3 rings) fused together (i.e., fused ring aryl) or covalently linked. Fused ring aryl refers to multiple rings fused together, wherein at least one fused ring is an aryl ring.

[0087] The term "heteroaryl" refers to an aryl group (or ring) containing one to four heteroatoms selected from N, O and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom is optionally quaternized. Therefore, the term "heteroaryl" includes fused ring heteroaryl groups (i.e., multiple rings fused together, wherein at least one of the fused rings is a heteroaromatic ring). 5,6-fused ring heteroarylene refers to two rings fused together, wherein one ring has 5 members and the other ring has 6 members, and wherein at least one ring is a heteroaryl ring. Similarly, 6,6-fused ring heteroarylene refers to two rings fused together, wherein one ring has 6 members, the other ring has 6 members, and wherein at least one ring is a heteroaryl ring. And 6,5-fused ring heteroarylene refers to two rings fused together, wherein one ring has 6 members, the other ring has 5 members, and wherein at least one ring is a heteroaryl ring. Heteroaryl can be connected to the rest of the molecule by carbon or heteroatom. Non-limiting examples of aryl and heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 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-isoxazolyl, 4-isoxazolyl, 5- isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl and 6-quinolyl. Substituents for each of the above mentioned aryl and heteroaryl ring systems are selected from the group consisting of the acceptable substituents described below.

[0088] "Arylene" and "heteroarylene" by themselves or as part of another substituent refer to a divalent radical derived from aryl and heteroaryl, respectively.

[0089] For brevity, the term "aryl" when used in combination with other terms (e.g., aryloxy, arylthio, aralkyl) includes aryl and heteroaryl rings as defined above. Thus, the term "aralkyl" is meant to include those groups in which an aryl group is attached to an alkyl group (e.g., benzyl, phenethyl, pyridylmethyl, etc.), including those alkyl groups in which a carbon atom (e.g., a methylene group) has been replaced, for example, by an oxygen atom (e.g., phenoxymethyl, 2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, etc.).

[0090] Each of the above terms (eg, "alkyl," "heteroalkyl," "aryl," and "heteroaryl") is meant to include both substituted and unsubstituted forms of the indicated radical. Preferred substituents for each radical are provided below.

[0091] As used herein, the term "heteroatom" or "ring heteroatom" is meant to include oxygen (O), nitrogen (N), sulfur (S), phosphorus (P) and silicon (Si).

[0092] Certain commonly used alternative chemical names may be used. For example, divalent groups, such as divalent "alkyl" groups, divalent "aryl" groups, etc., may also be referred to as "alkylene" groups or "alkylenyl" groups, "arylene" groups or "arylenyl" groups, respectively. In addition, unless otherwise expressly stated, when a combination of groups is referred to herein as a moiety, such as aralkyl, the last mentioned group includes the atoms by which the moiety is attached to the rest of the molecule.

[0093] The term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not occur. Likewise, the term "optionally substituted" means that any one or more hydrogen atoms on a designated atom or group may or may not be substituted with a moiety other than hydrogen.

[0094] Some compounds exist in tautomer forms. Tautomers are in balance with each other. For example, compounds containing amides can exist in balance with imidic acid (imidic acid) tautomers. No matter which tautomer is shown, and no matter how the equilibrium properties between the tautomers are, those of ordinary skill in the art will interpret the compound as both comprising amide tautomers and imidic acid tautomers. Therefore, compounds containing amides are interpreted as including their imidic acid tautomers. Equally, compounds containing imidic acid are interpreted as including their amide tautomers.

[0095] Any formula or structure given herein is also intended to represent unlabeled forms of the compound as well as isotopically labeled forms. An isotopically labeled compound has a structure described by the formula given herein except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into the disclosed compounds include 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 125I. Various isotopically labeled compounds of the present disclosure, for example, compounds in which radioactive isotopes such as 3 H. 13 C and 14 Such isotopically labeled compounds are useful in metabolic studies, reaction kinetic studies, detection or imaging techniques, such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including drug or substrate tissue distribution assays or radioisotope therapy of patients.

[0096] The present disclosure also includes "deuterated analogs" of compounds of Formula I, wherein 1 to n hydrogens attached to the carbon atom are replaced by deuterium, wherein n is the number of hydrogens in the molecule. Such compounds exhibit increased metabolic resistance and can therefore be used to increase the half-life of any compound of Formula I when administered to mammals, particularly humans. See, e.g., Foster, "Deuterium Isotope Effects in Studies of Drug Metabolism," Trends Pharmacol. Sci. 5(12): 524-527 (1984). Such compounds are synthesized by methods well known in the art, such as by using raw materials in which one or more hydrogens have been replaced by deuterium.

[0097] Deuterium-labeled or substituted therapeutic compounds of the present disclosure can have improved DMPK (drug metabolism and pharmacokinetics) properties, which relate to distribution, metabolism and excretion (ADME). Due to higher metabolic stability, such as longer in vivo half-life, reduced dosage requirements and / or an increase in therapeutic index, substitution with heavier isotopes such as deuterium can provide certain therapeutic advantages. 18 F-labeled compounds can be used for PET or SPECT studies. Isotope-labeled compounds and prodrugs thereof disclosed herein can generally be prepared by carrying out the processes disclosed in the schemes or examples and preparations described below, by 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.

[0098] The concentration of such heavier isotopes, particularly deuterium, can be defined by an isotopic enrichment factor. In the compounds of the present disclosure, 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", the position is understood to have hydrogen in its natural abundance isotopic composition. Therefore, in the compounds of the present disclosure, any atom specifically designated as deuterium (D) represents deuterium.

[0099] In many cases, the compounds of the present disclosure are capable of forming acid and / or base salts by virtue of the presence of amino and / or carboxyl groups or groups similar thereto.

[0100] Also provided are pharmaceutically acceptable salts, hydrates, solvates, tautomeric forms, 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.

[0101] The term "pharmaceutically acceptable salt" of a given compound refers to a salt that retains the biological effectiveness and properties of the given compound and is not biologically or otherwise undesirable. "Pharmaceutically acceptable salts" or "physiologically acceptable salts" include, for example, salts with inorganic acids and salts with organic acids. In addition, if the compounds described herein are obtained as acid addition salts, the free base can be obtained by alkalizing a solution of the acid salt. On the contrary, if the product is a free base, an addition salt, particularly a pharmaceutically acceptable addition salt, can be prepared according to conventional methods 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 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 acids and organic acids. Salts derived from inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. 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, salicylic acid, and the like. Similarly, pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases. By way of example only, salts derived from inorganic bases include sodium, potassium, lithium, ammonium, calcium, and magnesium salts. Salts derived 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), substituted alkylamines (i.e., NH2(substituted alkyl)), di(substituted alkyl)amines (i.e., HN(substituted alkyl)2), tri(substituted alkyl)amines (i.e., N(substituted alkyl)3), alkenylamines (i.e., NH2(alkenyl)), dienylamines (i.e., HN(alkenyl)2), trialnylamines (i.e., N(alkenyl)3), substituted alkenylamines (i.e., NH2(substituted alkenyl)), Di(substituted alkenyl)amines (i.e. HN(substituted alkenyl)2), tri(substituted alkenyl)amines (i.e. N(substituted alkenyl)3, mono-, di- or tri-cycloalkylamines (i.e. NH2(cycloalkyl), HN(cycloalkyl)2, N(cycloalkyl)3), mono-, di- or tri-arylamines (i.e. NH2(aryl), HN(aryl)2, N(aryl)3), or mixed amines, etc. Specific examples of suitable amines include, by way of example only, isopropylamine, trimethylamine, diethylamine, tri(isopropyl)amine, tri(n-propyl)amine, ethanolamine, 2-dimethylaminoethanol, piperazine, piperidine, morpholine, N-ethylpiperidine, and the like.

[0102] The term "substituted" refers to any one or more hydrogen atoms on a designated atom or group being replaced by one or more substituents other than hydrogen, provided that the normal valence of the designated atom is not exceeded. One or more substituents include, but are not limited to, alkyl, alkenyl, alkynyl, alkoxy, acyl, amino, amide, amidino, aryl, azido, carbamoyl, carboxyl, carboxyl ester, cyano, guanidino, halo, haloalkyl, haloalkoxy, heteroalkyl, heteroaryl, heterocyclic, hydroxyl, hydrazine, imino, oxo, nitro, alkylsulfinyl, sulfonic acid, alkylsulfonyl, thiocyanate, thiol, thioketone or a combination thereof. Polymers or similar indefinite structures (e.g., substituted aryls with substituted alkyls, substituted alkyls themselves being substituted by substituted aryls, further substituted by substituted heteroalkyls, etc.) obtained by limiting the unlimited addition of further substituents are not intended to be included herein. Unless otherwise indicated, the maximum number of consecutive substitutions in the compounds described herein is three. For example, the continuous substitution of substituted aryl by two other substituted aryl is limited to ((substituted aryl) substituted aryl) substituted aryl. Similarly, the above definition is not intended to include unallowed substitution patterns (e.g., methyl substituted by 5 fluorines or heteroaryl with two adjacent oxygen ring atoms). Such unallowed substitution patterns are well known to technicians. When used to modify chemical groups, the term "substituted" can describe other chemical groups defined herein. Unless otherwise specified, when a group is described as optionally substituted, any substituent of the group itself is unsubstituted. For example, in some embodiments, the term "substituted alkyl" refers to an alkyl with one or more substituents, and the substituents include hydroxyl, halogen, alkoxy, cycloalkyl, heterocyclic radical, aryl and heteroaryl. In other embodiments, one or more substituents can be further substituted by halogen, alkyl, haloalkyl, hydroxyl, alkoxy, cycloalkyl, heterocyclic radical, aryl or heteroaryl (each of which is substituted). In other embodiments, the substituents may be further substituted with halogen, alkyl, haloalkyl, alkoxy, hydroxy, cycloalkyl, heterocyclyl, aryl, or heteroaryl (each of which is unsubstituted).

[0103] As used herein, "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Unless any conventional media or agent is incompatible with the active ingredient, it is contemplated for use in the therapeutic compositions. Supplementary active ingredients may also be incorporated into the compositions.

[0104] A "solvate" is formed by the interaction of a solvent and a compound. Also provided are solvates of the salts of the compounds described herein. Also provided are hydrates of the compounds described herein.

[0105] Detection of Traumatic Brain Injury (TBI)

[0106] Consider traumatic brain injury (TBI), even mild TBI, which can rapidly cause the release and / or accumulation of beta-amyloid (Aβ) in the patient's retina. As shown in the experimental examples, such presence of Aβ in the retina can be detected with probes capable of binding to Aβ, and the binding can then be detected by means such as laser-activated retinal fluorescence scanning.

[0107] A "traumatic brain injury" (TBI) occurs when a bump, blow, jolt, or other injury to the head causes damage to the brain. TBI can cause permanent brain damage or death. Half of all TBIs result from motor vehicle accidents. Military personnel in combat zones are also at risk. Symptoms of a TBI may not appear until days or weeks after the injury. A concussion is the mildest type. It can cause headaches or neck pain, nausea, ringing in the ears, dizziness, and tiredness. People with moderate or severe TBI may experience symptoms such as headaches that get worse or don't go away, repeated vomiting or nausea, convulsions or seizures, inability to wake from sleep, slurred speech, weakness or numbness in the arms and legs, and dilated pupils.

[0108] "Amyloid beta" (Aβ or Abeta) refers to a peptide of about 36 to 43 amino acids associated with Alzheimer's disease (AD), as the main component of amyloid plaques found in the brains of Alzheimer's patients. The peptide is derived from amyloid precursor protein (APP; example GenBank accession number: NP_000475), which is cleaved by β-secretase and γ-secretase to produce Aβ. Aβ molecules can aggregate to form flexible soluble oligomers, which can exist in a variety of forms. It is believed that certain misfolded oligomers (called "seeds") can induce other Aβ molecules to also take the form of misfolded oligomers, resulting in a chain reaction similar to prion protein infection. Oligomers are toxic to nerve cells.

[0109] Therefore, according to one embodiment of the present disclosure, a method for determining whether a patient has suffered a traumatic brain injury (TBI) is provided. The method requires detecting the presence of amyloid beta protein in the patient's eye. Considering that the accumulation of amyloid beta protein mainly occurs in the retina. Therefore, the detection can mainly target the amyloid beta protein in the retina of the eye.

[0110] It is contemplated that the accumulation of amyloid beta may begin quickly following a physical impact to the head, which results in a TBI. It is understood that physical impact may not be the only cause of a TBI. Sound, light, and temperature may also result in a TBI and are therefore within the scope of the present disclosure. In some embodiments, the detection is performed within 1 or 2 hours after the cause of the potential TBI. Alternatively, the detection may be performed within 6, 12, 18, 24, 36, or 48 hours. In some embodiments, the detection is performed at least 1, 2, 3, 4, 6, 8, 12, 18, or 24 hours after the cause of the potential TBI. In some embodiments, the detection is performed at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 days after the cause of the potential TBI. In some embodiments, the detection is performed within 3, 4, 5, 6, 7, 8, 9, or 10 days after the cause of the potential TBI or within 1, 2, 3, 4, or 5 weeks.

[0111] In some embodiments, the patient may suffer from another disease or condition that causes amyloid beta protein accumulation. An example of such a disease is Alzheimer's disease (AD). However, it is contemplated that the patient receiving the detection method disclosed herein does not suffer from such other diseases or conditions (e.g., AD). In some embodiments, the patient is young enough (e.g., less than 70, 65, 60, 55, 50, 45, or 40 years old) so that it is not reasonably suspected that he or she suffers from such a disease or condition.

[0112] A direct physical impact to the eye (or light, sound or temperature) may cause amyloid beta protein to be released or accumulate in the eye. In some embodiments, the patient of the method is not subjected to such a direct impact to the eye.

[0113] In another embodiment, a method of preparing a patient for diagnosis of traumatic brain injury (TBI) is provided, the method comprising administering to the patient's eye a probe that specifically binds to amyloid beta protein. The types of patients suitable for this method are described above without limitation. Once the probe is administered to the patient's eye, its binding to amyloid beta protein can be detected using the methods described herein, and the binding indicates accumulation of amyloid beta protein, which is a sign of brain trauma.

[0114] Amyloid β Binding Probes and Assays

[0115] Detection of amyloid β protein can be performed using probes that can selectively bind to amyloid β protein, referred to herein as amyloid β binding (Aβ binding) probes. Such probes are known or can be easily developed or prepared.

[0116] In one embodiment, the Aβ binding probe can be detected by the fluorescent signal it emits when bound to Aβ after being activated by laser light. The following section provides examples of Aβ binding fluorescent probes.

[0117] The Aβ binding probe can also be an antibody that specifically binds to Aβ. Antibodies against proteins or peptides can be routinely developed and prepared from animal sources or by methods such as phage display. Detection of antibodies can also be performed by methods known in the art, such as probes that can bind to antibodies and can emit detectable signals. Non-limiting examples of antibodies include the 6E10 antibody used herein and ab2539 available from Abcam (Cambridge, MA).

[0118] A retinal imaging device can be used to facilitate in situ detection of the binding of the Aβ binding probe to Aβ in the patient's retina, and the retinal imaging device is preferably handheld or portable. The retinal imaging device may include a lens and an image sensor, and optionally a laser light source. When the light source emits laser light to the retina, if Aβ accumulates on the retina and binds to the Aβ binding probe, the accumulation can be easily detected and quantified by the lens and image sensor that collect and sense the fluorescent signal.

[0119] Amyloid-sensitive fluorescent probe (ASF)

[0120] The present disclosure further provides a small molecule fluorescent probe capable of binding to amyloid β (Aβ). In one embodiment, the probe can be selected from WO 2011 / 072257, WO 2015 / 143185 or WO

[0121] 2017 / 004560, which is incorporated herein by reference in its entirety. In one embodiment, the probe can be curcumin or other compounds currently used to stain Aβ, such as thioflavin or Congo red.

[0122] In one embodiment, the probe may be selected from the compounds described in WO 2015 / 143185, which are described below.

[0123] In certain embodiments, the present disclosure provides a compound of Formula I, or a salt or solvate thereof:

[0124]

[0125] in

[0126] EDG is an electron donating group;

[0127] Each Ar is independently C1-C 14 Arylene or C1-C 14 heteroarylene, each optionally substituted with one or more R1;

[0128] Each R1 is independently halogen, -OR2, -NR3R4, C 1- C 10Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 heteroarylene;

[0129] wherein alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted with one or more R5;

[0130] R2, R3 and R4 are independently hydrogen, C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 heteroarylene, each of which, except hydrogen, is optionally substituted with one or more R5;

[0131] Each R5 is independently halogen, -OR6, -NR7R8, C 1- C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 heteroarylene;

[0132] R6, R7, R8 and R 84 are independently hydrogen or C1-C 10 alkyl;

[0133] R 84 is hydrogen or C1-C 10 alkyl;

[0134] EWG is an electron withdrawing group;

[0135] WSG is a water-soluble group;

[0136] X is C=O or SO2 or X and R 84 Combine to form pyridyl;

[0137] Y is NH or S;

[0138] Each x is independently an integer from 0 to 10;

[0139] Each w is independently an integer from 1 to 5;

[0140] Each y is independently an integer from 0 to 10; and

[0141] z is an integer from 1 to 10.

[0142] In certain embodiments, the compound is a compound of Formula II or a salt or solvate thereof:

[0143]

[0144] in

[0145] EDG is an electron donating group;

[0146] Ar2 and each Ar1 are independently C1-C 14 Arylene or C1-C 14 heteroarylene;

[0147] Each optionally represented by one or more R 41 replace;

[0148] Each R 41 are independently halogen, -CN, -OR 42 、-NR 43 R 44 、C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 wherein alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted by one or more R 45 replace;

[0149] R 42 , R 43 and R 44 are independently hydrogen, C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 Heteroarylene, each of which, except hydrogen, is optionally substituted by one or more R 45 replace;

[0150] Each R 45 are independently halogen, -OR 46 、-NR 47 R 48 、C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C10 Heterocycloalkyl, C1-C 10 Arylene, or C1-C 10 heteroarylene; and

[0151] R 46 , R 47 and R 48 are independently hydrogen or C1-C 10 alkyl;

[0152] EWG is an electron withdrawing group;

[0153] Y is absent, O, NH or S;

[0154] WSG is hydrogen or water-soluble group;

[0155] x is an integer from 0 to 10;

[0156] y is an integer between 0 and 10; and

[0157] z is an integer from 1 to 10.

[0158] In certain embodiments, R 84 In certain embodiments, R 84 C1-C 10 In certain embodiments, R 84 is methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, heptyl or decyl. In certain embodiments, R 84 It is methyl.

[0159] As is known in the art, the substituent EDG is an electron donor group. In certain embodiments, EDG is any atom or functional group that is capable of donating some of its electron density to a conjugated π system, thereby making the π system more nucleophilic.

[0160] In certain embodiments,

[0161] EDG is -OR9, -NR 10 R 11 、-SR 12 , -PR 13 R 14 、-NR 15 C(O)R 16 、C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 Heteroarylene,

[0162] wherein alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted by one or more R 17 replace;

[0163] Each R 17 are independently halogen, -OR 18 、-NR 19 R 20 、C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene, or C1-C 10 heteroarylene;

[0164] R9, R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 18 , R 19 and R 20 Each of which is independently hydrogen, C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 Heteroarylene,

[0165] Each of these, except hydrogen, is optionally replaced by one or more R 21 substituted, and wherein R 10 and R 11 optionally linked together to form optionally R 21 substituted heterocycloalkyl or heteroaryl;

[0166] Each R 21 are independently halogen, -OR 22 、-NR 23 R 24 、C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene, or C1-C 10 Heteroarylene,

[0167] wherein alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted by one or more R 25 replace;

[0168] R 22 , R 23 and R 24 Each of 10 Alkyl; and

[0169] Each R 25 Independently C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 Heteroarylene.

[0170] In certain embodiments, the EDG is selected from:

[0171]

[0172] In certain embodiments, EDG is In certain embodiments, EDG is

[0173]

[0174] EWG is an electron withdrawing group. In certain embodiments, an electron withdrawing group as used herein is any atom or group that is capable of withdrawing electron density from neighboring atoms toward itself through resonance or inductive effects.

[0175] In certain embodiments,

[0176] EWG is selected from halogen, -CN, -NO2, -SO3H, -CR 26 R 27 R 28 、-COR 29 or -COOR 30 ;

[0177] Each R 26 , R 27 and R 28 are independently hydrogen or halogen;

[0178] R 29 Halogen, hydrogen, C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10Arylene or C1-C 10 Heteroarylene,

[0179] wherein alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted by one or more R 31 replace;

[0180] R 30 is hydrogen, C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 Heteroarylene,

[0181] wherein alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted by one or more R 32 Replacement; and

[0182] Each R 31 and R 32 Independently C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 Heteroarylene.

[0183] In certain embodiments, EWG is selected from -F, -Cl, -Br, -CH=O, NO2, -CF3, -CCl3, -SO3, and -CN. In certain embodiments, EWG is F, Cl, or Br. In certain embodiments, EWG is -CN.

[0184] WSG is a water-soluble group. In certain embodiments, the WSG group is used to alter the solubility of a compound in an aqueous system.

[0185] In certain embodiments,

[0186] WSG is hydrogen, C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 Heteroarylene,

[0187] wherein alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted by one or more R 33 replace;

[0188] Each R 33 are independently halogen, -OR 34 、-NR 35 R 36 、C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 Heteroarylene,

[0189] wherein alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted by one or more R 37 replace;

[0190] Each R 34 , R 35 and R 36 are independently hydrogen, C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 Heteroarylene,

[0191] wherein alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted by one or more R 37 replace;

[0192] Each R 37 are independently halogen, -OR 38 、-NR 39 R 40 、C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, -(C 1- C6 alkyl)(C1-C 10 Heterocycloalkylene), C1-C 10 Arylene or C1-C 10 heteroarylene; and

[0193] R 38 , R 39 and R 40 Each of 10 alkyl.

[0194] In certain embodiments, WSG is

[0195] In certain embodiments, the WSG is polyethylene glycol, polypropylene glycol, a copolymer of polyethylene glycol and polypropylene glycol, or an alkoxy derivative thereof.

[0196] In certain embodiments, WSG is Where n is an integer from 1 to 50, R 81 is hydrogen, C1-C 10 Alkyl, C1-C 10 Alkenyl or C1-C 10 Alkynyl, wherein each alkyl, alkenyl or alkynyl group is optionally substituted with one or more C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 In certain embodiments, R 81 In certain embodiments, R 81 In certain embodiments, R 81 In certain embodiments, R 81 In certain embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50. In certain embodiments, n is an integer of value 1-10, 1-20, 1-30, 1-40, 1-50, 10-20, 10-30, 10-40, 10-50, 20-30, 20-40, 20-50, 30-40, 30-50, or 40-50. In certain embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In certain embodiments, n is 3 or 6.

[0197] In certain embodiments, R 81 For hydrogen.

[0198] In certain embodiments, WSG is

[0199] In certain embodiments, WSG is

[0200] In certain embodiments, WSG is

[0201] In certain embodiments, WSG is

[0202] In certain embodiments, WSG is Each R 82 are independently hydrogen or C1-C 10 alkyl.

[0203] In certain embodiments, each R 82 are independently hydrogen, methyl, ethyl, propyl or butyl.

[0204] In certain embodiments, WSG is

[0205]

[0206] In certain embodiments, WSG is

[0207]

[0208] In certain embodiments, WSG is Each R 83 is hydrogen or C1-C 10 In certain embodiments, each R 83 are independently hydrogen, methyl, ethyl, propyl or butyl.

[0209] In certain embodiments, WSG is

[0210]

[0211] In certain embodiments, WSG is

[0212]

[0213] In certain embodiments, WSG is -(C1-C 10 Alkylene)-R 33 -R 37 In certain embodiments, WSG is -(C1-C 10 Alkylene)-R 33 -R 37 And R 33 C1-C 10 In certain embodiments, WSG is -(C1-C 10 Alkyl)-R 33 -R 37 , R 33 C1-C 10 heteroarylene, and R 37 is -(C1-C6 alkyl)(C1-C10) In certain embodiments, WSG is -CH2-R 33 -R 37 In certain embodiments, WSG is -CH2-R 33 -R 37 And R 33 is triazole, imidazole or pyrazole. In certain embodiments, WSG is -CH2-R 33 -R 37 And R 33 In certain embodiments, WSG is

[0214] –CH2-R 33 -R 37 And R 33 is 1,2,4-triazole. In certain embodiments, WSG is -CH2-R 33 -R 37 And R 33 is 1,2,3-triazole. In certain embodiments, WSG is -CH2-R 33 -R 37 , R 33 is 1,2,3-triazole, and R 37 is -(C1-C6 alkyl)(C1-C 10 In certain embodiments, WSG is -CH2-R 33 -R 37 , R 33 is 1,2,3-triazole, and R 37 is -(C1 alkyl)(C1-C 10 In certain embodiments, WSG is -CH2-R 33 -R 37 , R 33 is 1,2,3-triazole, R 37 is -(C1 alkyl)(C1-C 10 Heterocycloalkyl), and C1-C 10 Heterocycloalkyl is a tetrahydropyran derivative.

[0215] In certain embodiments, WSG is Each R 87 is hydrogen, C1-C 10 Alkyl or -C(=O)C1-C 10 In certain embodiments, each R 87 R is independently hydrogen, methyl, ethyl, propyl, butyl, acetate, propionate or butyrate. 87 is independently hydrogen or methyl. In certain embodiments, each R 87is independently methyl or acetate.

[0216] In certain embodiments, WSG is

[0217] In certain embodiments, WSG is

[0218] In certain embodiments, WSG is -(C1-C 10 Heteroalkyl)-R 33 -R 37 In certain embodiments, WSG is -(C1-C 10 Heteroalkyl)-R 33 -R 37 And R 33 C1-C 10 In certain embodiments, WSG is -(C1-C 10 Heteroalkyl)-R 33 -R 37 And R 33 C1-C 10 heteroarylene, and R 37 is -(C1-C6 alkyl)(C1-C 10 heterocycloalkyl).

[0219] In certain embodiments, WSG is and p is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50. In certain embodiments, p is an integer having a value of 1-10, 1-20, 1-30, 1-40, 1-50, 10-20, 10-30, 10-40, 10-50, 20-30, 20-40, 20-50, 30-40, 30-50, or 40-50. In certain embodiments, p is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In certain embodiments, p is 3 or 6. In certain embodiments, p is 3.

[0220] In certain embodiments, WSG is And R 33 C1-C 10 In certain embodiments, R 33 is C5 heteroarylene. In certain embodiments, R 33 is triazole, imidazole or pyrazole. 33In certain embodiments, R 33 is 1,2,4-triazole. In certain embodiments, R 33 is 1,2,3-triazole. In certain embodiments, R 33 is 1,2,3-triazole, and p is 3. In certain embodiments, R 33 is 1,2,3-triazole, and R 37 is -(C1-C6 alkyl)(C1-C 10 In certain embodiments, R 33 is 1,2,3-triazole, and R 37 is -(C1 alkyl)(C1-C 10 In certain embodiments, R 33 is 1,2,3-triazole, R 37 is a tetrahydropyran derivative. In certain embodiments, R 33 is 1,2,3-triazole, and R 37 for

[0221] In certain embodiments, WSG is R 33 is 1,2,3-triazole, R 37 for p is 3.

[0222] In certain embodiments, WSG is Each R 87 is hydrogen, C1-C 10 Alkyl or -C(=O)C1-C 10 In certain embodiments, each R 87 R is independently hydrogen, methyl, ethyl, propyl, butyl, acetate, propionate or butyrate. 87 is independently hydrogen or methyl. In certain embodiments, each R 87 is independently methyl or acetate.

[0223] In certain embodiments, WSG is

[0224] In certain embodiments, WSG is

[0225] In certain embodiments, X is C=O or SO2. In certain embodiments, X is C=O. In certain embodiments, X is SO2.

[0226] In certain embodiments, Y is NH or S. In certain embodiments, Y is NH. In certain embodiments, Y is S.

[0227] The variable w in Formula I is an integer from 1 to 5. In certain embodiments, w is 1. In certain embodiments, w is 2. In certain embodiments, w is 3. In certain embodiments, w is 4. In certain embodiments, w is 5.

[0228] The variable x in Formula I is an integer from 0 to 10. In certain embodiments, x is 0. In certain embodiments, x is 1. In certain embodiments, x is 2. In certain embodiments, x is 3. In certain embodiments, x is 4. In certain embodiments, x is 5. In certain embodiments, x is 6. In certain embodiments, x is 7. In certain embodiments, x is 8. In certain embodiments, x is 9. In certain embodiments, x is 10.

[0229] The variable y in Formula I is an integer from 0 to 10. In certain embodiments, y is 0. In certain embodiments, y is 1. In certain embodiments, y is 2. In certain embodiments, y is 3. In certain embodiments, y is 4. In certain embodiments, y is 5. In certain embodiments, y is 6. In certain embodiments, y is 7. In certain embodiments, y is 8. In certain embodiments, y is 9. In certain embodiments, y is 10.

[0230] The variable z in Formula I is an integer from 1 to 10. In certain embodiments, z is 1. In certain embodiments, z is 2. In certain embodiments, z is 3. In certain embodiments, z is 4. In certain embodiments, z is 5. In certain embodiments, z is 6. In certain embodiments, z is 7. In certain embodiments, z is 8. In certain embodiments, z is 9. In certain embodiments, z is 10.

[0231] In certain embodiments, x is 0, w is 1, y is 0, z is 1, X is C═O, and Y is NH.

[0232] In certain embodiments, x is 0, w is 1, y is 0, z is 1, X is SO2, and Y is NH.

[0233] In certain embodiments, x is 0, w is 2, y is 0, z is 1, X is C=O, and Y is NH.

[0234] In certain embodiments, x is 0, w is 2, y is 0, z is 1, X is SO2, and Y is NH.

[0235] In certain embodiments, the present disclosure provides compounds of Formula Ia:

[0236] Among them, EDG, Ar, R 84, x, w, y, z, EWG and WSG are defined as described above.

[0237] In certain embodiments, the present disclosure provides compounds of Formula Ib:

[0238] Among them, EDG, Ar, R 84 , x, w, y, z, EWG and WSG are defined as described above.

[0239] In one aspect, the present disclosure provides compounds of Formula Ic:

[0240] Among them, EDG, Ar, R 84 , X, Y, EWG and WSG are defined as described above.

[0241] In certain embodiments, the compound of formula (Ic) or a salt or solvate thereof

[0242]

[0243] in

[0244] EDG is:

[0245] a) a heterocycloalkyl group of not more than 10 carbon atoms, which is optionally substituted by one or more R 17 replace; or

[0246] b)-NR 10 R 11 ;

[0247] Each R 17 are independently halogen, -OR 18 、-NR 19 R 20 、C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons;

[0248] R 10 , R 11 , R 18 , R 19 and R 20 Each of which is independently hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbon atoms, heterocycloalkyl of up to 10 carbon atoms, arylene of up to 10 carbon atoms, or heteroarylene of up to 10 carbon atoms, each of which is optionally replaced by one or more R 21 replace;

[0249] R 21 Each of which is independently halogen, -OR 22 、-NR 23 R 24 、C1-C 10 Alkyl, C1-C 10 is a heteroalkyl, a cycloalkyl of up to 10 carbon atoms, a heterocycloalkyl of up to 10 carbon atoms, an arylene of up to 10 carbon atoms, or a heteroarylene of up to 10 carbon atoms, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally replaced by one or more R 25 replace;

[0250] R 22 , R 23 and R 24 Each of 10 Alkyl; and

[0251] Each R 25 Independently C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons;

[0252] Ar is an arylene group having not more than 14 carbon atoms or a heteroarylene group having not more than 14 carbon atoms,

[0253] each optionally substituted with one or more R1;

[0254] Each of R1 is independently halogen, -OR2, -NR3R4, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted with one or more R5;

[0255] R2, R3 and R4 are independently hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons, each of which is optionally substituted with one or more R5, excluding hydrogen;

[0256] Each R5 is independently halogen, -OR6, -NR7R8, C1-C 10 Alkyl, C1-C 10heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons;

[0257] R6, R7, R8 and R 84 are independently hydrogen or C1-C 10 alkyl;

[0258] EWG is selected from -F, -Cl, -Br, -CH=O, NO2, -CF3, -CCl3, -SO3H and -CN;

[0259] WSG is:

[0260] i)

[0261]

[0262] ii) polyethylene glycol, polypropylene glycol, copolymers of polyethylene glycol and polypropylene glycol or alkoxy derivatives thereof;

[0263] iii)

[0264]

[0265] Where n is an integer from 1 to 50 and R 81 is hydrogen, C1-C 10 Alkyl, C1-C 10 Alkenyl or C1-C 10 Alkynyl, wherein the alkyl, alkenyl or alkynyl group is optionally substituted with one or more C1-C 10 Alkyl, C1-C 10 substituted with heteroalkyl, cycloalkyl of up to 10 carbon atoms, heterocycloalkyl of up to 10 carbon atoms, arylene of up to 10 carbon atoms, or heteroarylene of up to 10 carbon atoms;

[0266] iv)

[0267]

[0268] v)

[0269]

[0270] vi)

[0271] -(C1-C 10 Alkyl)-R 33 -R 37 ,in:

[0272] R 33 is a heteroarylene group having no more than 10 carbon atoms; and

[0273] R 37is -(C1-C6 alkyl)(heterocycloalkyl with no more than 10 carbon atoms); vii)

[0274]

[0275] viii)

[0276] –(C1-C 10 Heteroalkyl)-R 33 -R 37 ,in:

[0277] R 33 is a heteroarylene group having no more than 10 carbon atoms; and

[0278] R 37 is -(C1-C6 alkyl)(heterocycloalkyl not exceeding 10 carbon atoms); or ix)

[0279]

[0280] X is C=O or SO2 or X and R 84 Combine to form pyridyl;

[0281] Y is NH or S.

[0282] In certain embodiments, R 81 For hydrogen.

[0283] In certain embodiments, WSG is In certain embodiments, WSG is

[0284] In one aspect, the present disclosure provides compounds of Formula Id:

[0285] Among them, EDG, R 84 , Ar, EWG and WSG are as defined above.

[0286] In one aspect, the present disclosure provides compounds of Formula Ie:

[0287] Among them, EDG, R 84 , Ar, EWG and WSG are as defined above.

[0288] In some cases, the compound is selected from:

[0289]

[0290]

[0291] wherein n is an integer having a value of 1 to 50. In some cases, n is an integer having a value of 1 to 10, for example, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0292] In certain embodiments, the compound is selected from:

[0293]

[0294]

[0295]

[0296] In certain embodiments, the compound is selected from:

[0297]

[0298]

[0299] In certain embodiments, the compound is selected from the group consisting of:

[0300]

[0301]

[0302] wherein p is an integer having a value of 1 to 50. In certain instances, p is an integer having a value of 1 to 10, for example, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0303] In certain embodiments, the compound is selected from:

[0304]

[0305] wherein n is an integer having a value of 1 to 50. In some cases, n is an integer having a value of 1 to 10, for example, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0306] In certain embodiments, the compound is selected from:

[0307]

[0308]

[0309] In certain embodiments, the compound is selected from:

[0310]

[0311]

[0312] In certain embodiments, the compound is selected from the group consisting of:

[0313]

[0314]

[0315] In certain embodiments, the compound is selected from:

[0316]

[0317]

[0318] In certain embodiments, the compound is selected from:

[0319] wherein n is an integer having a value of 1 to 50. In some cases, n is an integer having a value of 1 to 10, for example, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0320] In certain embodiments, the compound is selected from:

[0321]

[0322] wherein n is an integer having a value of 1 to 50. In some cases, n is an integer having a value of 1 to 10, for example, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0323] In certain embodiments, the compound is selected from:

[0324] wherein n is an integer having a value of 1 to 50. In some cases, n is an integer having a value of 1 to 10, for example, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0325] In certain embodiments, the compound is selected from:

[0326] wherein n is an integer having a value of 1 to 50. In some cases, n is an integer having a value of 1 to 10, for example, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0327] In certain embodiments, the compound is selected from:

[0328]

[0329] wherein n is an integer having a value of 1 to 50. In some cases, n is an integer having a value of 1 to 10, for example, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0330] In certain embodiments, the compound is selected from:

[0331]

[0332] wherein n is an integer having a value of 1 to 50. In some cases, n is an integer having a value of 1 to 10, for example, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0333] In certain embodiments, the compound is selected from:

[0334]

[0335] In certain embodiments, the compound is selected from:

[0336]

[0337] In certain embodiments, the compound is selected from:

[0338]

[0339]

[0340] In certain embodiments, the compound is selected from:

[0341]

[0342] In certain embodiments, the compound is selected from:

[0343]

[0344] In certain embodiments, the compound is selected from:

[0345]

[0346]

[0347] In certain embodiments, the compound is selected from:

[0348]

[0349]

[0350] In certain embodiments, the compound is selected from:

[0351]

[0352]

[0353] In certain embodiments, the compound is selected from:

[0354]

[0355]

[0356] In certain embodiments, the compound is selected from:

[0357]

[0358]

[0359] In certain embodiments, the compound is selected from:

[0360]

[0361]

[0362] In certain embodiments, the compound is selected from:

[0363]

[0364]

[0365] In certain embodiments, the compound is selected from:

[0366]

[0367]

[0368] In certain embodiments, the compound is selected from:

[0369]

[0370] In certain embodiments, the compound is selected from:

[0371]

[0372] In certain embodiments, the compound is selected from:

[0373]

[0374]

[0375] In certain embodiments, the compound is selected from:

[0376]

[0377]

[0378] In certain embodiments, the compound is selected from:

[0379]

[0380]

[0381] In certain embodiments, the compound is

[0382] In certain embodiments, the compound is (E)-2-cyano-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-3-(6-(piperidin-1-yl)naphthalen-2-yl)acrylamide. In certain embodiments, the compound is (Z)-2-cyano-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-3-(6-(piperidin-1-yl)naphthalen-2-yl)acrylamide.

[0383] In certain embodiments, the compound is:

[0384]

[0385] In certain embodiments, the compound is (E)-1-cyano-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)ethylenesulfonamide. In certain embodiments, the compound is (Z)-1-cyano-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)ethylenesulfonamide.

[0386] In certain embodiments, the compound is

[0387]

[0388] In certain embodiments, the compound is (E)-2-cyano-N-(2,5,8,11,14,17-hexaoxa nonadecan-19-yl)-3-(6-(piperidin-1-yl)naphthalen-2-yl)acrylamide. In certain embodiments, the compound is (Z)-2-cyano-N-(2,5,8,11,14,17-hexaoxa nonadecan-19-yl)-3-(6-(piperidin-1-yl)naphthalen-2-yl)acrylamide.

[0389] In certain embodiments, the compound is

[0390]

[0391] In certain embodiments, the compound is (E)-1-cyano-N-(2,5,8,11,14,17-hexaoxa nonadecan-19-yl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)ethylenesulfonamide. In certain embodiments, the compound is (Z)-1-cyano-N-(2,5,8,11,14,17-hexaoxa nonadecan-19-yl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)ethylenesulfonamide.

[0392] In certain embodiments, the compound is

[0393]

[0394] In certain embodiments, the compound is (E)-2-cyano-N-(2,3-dihydroxypropyl)-3-(6-(piperidin-1-yl)naphthalen-2-yl)acrylamide. In certain embodiments, the compound is (Z)-2-cyano-N-(2,3-dihydroxypropyl)-3-(6-(piperidin-1-yl)naphthalen-2-yl)acrylamide.

[0395] In certain embodiments, the compound is:

[0396]

[0397] In certain embodiments, the compound is (E)-1-cyano-N-(2,3-dihydroxypropyl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)ethylenesulfonamide. In certain embodiments, the compound is (Z)-1-cyano-N-(2,3-dihydroxypropyl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)ethylenesulfonamide.

[0398] In certain embodiments, the compound is:

[0399]

[0400] In certain embodiments, the compound is (E)-2-cyano-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-3-(6-(piperidin-1-yl)naphthalen-2-yl)but-2-enamide. In certain embodiments, the compound is (Z)-2-cyano-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-3-(6-(piperidin-1-yl)naphthalen-2-yl)but-2-enamide.

[0401] In certain embodiments, the compound is:

[0402]

[0403] In certain embodiments, the compound is (E)-1-cyano-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)prop-1-ene-1-sulfonamide. In certain embodiments, the compound is (Z)-1-cyano-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)prop-1-ene-1-sulfonamide.

[0404] In certain embodiments, the compound is

[0405]

[0406] In certain embodiments, the compound is (E)-2-cyano-N-(2,5,8,11,14,17-hexaoxa nonadecan-19-yl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)but-2-enamide. In certain embodiments, the compound is (Z)-2-cyano-N-(2,5,8,11,14,17-hexaoxa nonadecan-19-yl)-3-(6-(piperidin-1-yl)naphthalen-2-yl)but-2-enamide.

[0407] In certain embodiments, the compound is:

[0408]

[0409] In certain embodiments, the compound is (E)-1-cyano-N-(2,5,8,11,14,17-hexaoxa nonadecan-19-yl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)prop-1-ene-1-sulfonamide. In certain embodiments, the compound is (Z)-1-cyano-N-(2,5,8,11,14,17-hexaoxa nonadecan-19-yl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)prop-1-ene-1-sulfonamide.

[0410] In certain embodiments, the compound is:

[0411]

[0412] In certain embodiments, the compound is (E)-2-cyano-N-(2,3-dihydroxypropyl)-3-(6-(piperidin-1-yl)naphthalen-2-yl)but-2-enamide. In certain embodiments, the compound is (Z)-2-cyano-N-(2,3-dihydroxypropyl)-3-(6-(piperidin-1-yl)naphthalen-2-yl)but-2-enamide.

[0413] In certain embodiments, the compound is:

[0414]

[0415] In certain embodiments, the compound is (E)-1-cyano-N-(2,3-dihydroxypropyl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)prop-1-ene-1-sulfonamide. In certain embodiments, the compound is (Z)-1-cyano-N-(2,3-dihydroxypropyl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)prop-1-ene-1-sulfonamide.

[0416] In certain embodiments, the compound is

[0417]

[0418] In certain embodiments, the compound is (R,E)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl

[0419] In certain embodiments, the compound is (R,Z)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl)-N-((3,4,5,6-tetrahydroxytetrahydro-2H-pyran-2-yl)methyl)acrylamide.

[0420] In certain embodiments, the compound is:

[0421]

[0422] In certain embodiments, the compound is (E)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl

[0423] )-N-(((2R,3S,4S,5R)-3,4,5,6-tetrahydroxytetrahydro-2H-pyran-2-yl)methyl)acrylamide. In certain embodiments, the compound is (Z)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl

[0424] )-N-(((2R,3S,4S,5R)-3,4,5,6-tetrahydroxytetrahydro-2H-pyran-2-yl)methyl)acrylamide.

[0425] In certain embodiments, the compound is:

[0426]

[0427] In certain embodiments, the compound is (E)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl

[0428] In certain embodiments, the compound is (Z)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl)-N-(2,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acrylamide.

[0429] In certain embodiments, the compound is:

[0430]

[0431] In certain embodiments, the compound is (E)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl

[0432] )-N-((3R,4R,5S,6R)-2,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acrylamide. In certain embodiments, the compound is (Z)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl

[0433] )-N-((3R,4R,5S,6R)-2,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acrylamide.

[0434] In certain embodiments, the compound is:

[0435]

[0436] In certain embodiments, the compound is (E)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl

[0437] In certain embodiments, the compound is (Z)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl)-N-((1-((3,4,5-trihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)methyl)-1H-1,2,3-triazol-4-yl)methyl)acrylamide.

[0438] In certain embodiments, the compound is:

[0439]

[0440] In certain embodiments, the compound is (E)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl

[0441] )-N-((1-(((2R,3S,4S,5R,6S)-3,4,5-trihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)methyl

[0442] In certain embodiments, the compound is (Z)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl)-N-((1-(((2R,3S,4S,5R,6S)-3,4,5-trihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)methyl)-1H-1,2,3-triazol-4-yl)methyl)acrylamide.

[0443] In certain embodiments, the compound is:

[0444]

[0445] In certain embodiments, the compound is (E)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl

[0446] In certain embodiments, the compound is (Z)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl)-N-(2-(2-(2-((1-((3,4,5-trihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)methyl)-1H-1,2,3-triazol-4-yl)methoxy)ethoxy)ethoxy)ethyl)acrylamide.

[0447] In certain embodiments, the compound is:

[0448]

[0449] In certain embodiments, the compound is (E)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl

[0450] )-N-(2-(2-(2-((1-(((2R,3S,4S,5R,6S)-3,4,5-trihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)methyl)-1H-1,2,3-triazol-4-yl)methoxy)ethoxy)ethoxy)ethyl)acrylamide. In certain embodiments, the compound is (Z)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl

[0451] )-N-(2-(2-(2-((1-(((2R,3S,4S,5R,6S)-3,4,5-trihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)methyl)-1H-1,2,3-triazol-4-yl)methoxy)ethoxy)ethoxy)ethyl)acrylamide.

[0452] In certain embodiments, the compound is

[0453] In certain embodiments, the compound is a pharmaceutically acceptable salt or solvate of Compound 21.

[0454] In certain embodiments, the compound is

[0455] In certain embodiments, the compound is a pharmaceutically acceptable salt or solvate of Compound 22.

[0456] In certain embodiments, the compound is

[0457]

[0458] In certain embodiments, each Ar1 is independently substituted or unsubstituted naphthylene or substituted or unsubstituted phenylene. In certain embodiments, Ar2 is substituted or unsubstituted naphthylene or substituted or unsubstituted phenylene. In certain embodiments, Ar2 is substituted or unsubstituted pyridyl, substituted or unsubstituted pyrimidyl, substituted or unsubstituted pyrazinyl or substituted or unsubstituted pyridazinyl. In certain embodiments, Ar2 is substituted or unsubstituted pyridyl.

[0459] The substituent EDG in Formula II is an electron donating group. In certain embodiments, EDG is any electron donating group known in the art. In certain cases, any atom or functional group is capable of donating some of its electron density to the conjugated pi system by resonance or induced electron withdrawal, thereby making the pi system more nucleophilic. In certain embodiments, EDG is -OR 49 、-NR 50 R 51 、-SR 52 , -PR 53 R 54 、-NR 55 C(O)R 56 、C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 Heteroarylene, wherein alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted by one or more R 57 Replace; wherein each R 57 are independently halogen, -OR 58 、-NR 59 R 60 、C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene, or C1-C 10 Heteroarylene; R 49 , R 50 , R 51 , R 52 , R 53 , R 54 , R 55 , R 56 , R 58 , R 59 and R 60 Each of which is independently hydrogen, C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 Heteroarylene, each of which, except hydrogen, is optionally substituted by one or more R 61 substituted, and wherein R 50 and R 51optionally linked together to form optionally R 61 substituted heterocycloalkyl or heteroaryl; each R 61 are independently halogen, -OR 62 、-NR 63 R 64 、C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene, or C1-C 10 Heteroarylene, wherein alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted by one or more R 65 Replacement; R 62 , R 63 and R 64 Each of 10 alkyl; and each R 65 Independently C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene or C1-C 10 Heteroarylene.

[0460] In certain embodiments, Y is absent, O, NH, or S. In certain embodiments, Y is absent (i.e., Y is a bond). In certain embodiments, Y is O. In certain embodiments, Y is NH. In certain embodiments, Y is S.

[0461] The variable x in Formula II is an integer from 0 to 10. In certain embodiments, x is 0. In certain embodiments, x is 1. In certain embodiments, x is 2. In certain embodiments, x is 3. In certain embodiments, x is 4. In certain embodiments, x is 5. In certain embodiments, x is 6. In certain embodiments, x is 7. In certain embodiments, x is 8. In certain embodiments, x is 9. In certain embodiments, x is 10.

[0462] The variable y in Formula II is an integer from 0 to 10. In certain embodiments, y is 0. In certain embodiments, y is 1. In certain embodiments, y is 2. In certain embodiments, y is 3. In certain embodiments, y is 4. In certain embodiments, y is 5. In certain embodiments, y is 6. In certain embodiments, y is 7. In certain embodiments, y is 8. In certain embodiments, y is 9. In certain embodiments, y is 10.

[0463] The variable z in Formula II is an integer from 1 to 10. In certain embodiments, z is 1. In certain embodiments, z is 2. In certain embodiments, z is 3. In certain embodiments, z is 4. In certain embodiments, z is 5. In certain embodiments, z is 6. In certain embodiments, z is 7. In certain embodiments, z is 8. In certain embodiments, z is 9. In certain embodiments, z is 10.

[0464] In certain embodiments, x is 0, y is 0, z is 1, and Y is O.

[0465] In certain embodiments, x is 0, y is 0, z is 1, and Y is S.

[0466] In certain embodiments, x is 0, y is 0, z is 1, and Y is NH.

[0467] In certain embodiments, x is 0, y is 0, z is 1, and Y is absent.

[0468] In certain embodiments, x is 0, y is 0, z is 2, and Y is O.

[0469] In certain embodiments, x is 0, y is 0, z is 2, and Y is S.

[0470] In certain embodiments, x is 0, y is 0, z is 2, and Y is NH.

[0471] In certain embodiments, x is 0, y is 0, z is 2, and Y is absent.

[0472] In one aspect, the present disclosure provides compounds of Formula IIa:

[0473] wherein EDG, Ar1, Ar2, Y, EWG and WSG are as defined in Formula II above.

[0474] In one aspect, the present disclosure provides compounds of Formula IIb:

[0475] wherein EDG, Ar1, Ar2, Y, EWG and WSG are as defined in Formula II above.

[0476] In certain cases, the compound according to Formula II is selected from:

[0477]

[0478] wherein n is an integer having a value of 0 to 50. In certain embodiments, n is an integer having a value of 0 to 10, for example, n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0479] In certain embodiments, the compound is selected from:

[0480]

[0481]

[0482] In certain embodiments, the compound is selected from:

[0483]

[0484]

[0485]

[0486] Where R 85 Is H or CN. In certain embodiments, the compound is selected from:

[0487]

[0488]

[0489] Where R 85 is H or CN, R 86 for wherein n is an integer with a value of 0 to 50. In certain embodiments, n is an integer with a value of 0 to 10, for example, n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0490] In certain embodiments, the compound is selected from:

[0491]

[0492] Where R 85 is H or CN and R 86 For H.

[0493] In certain embodiments, the compound is selected from:

[0494] Where R 85 is H or CN and R 86 for wherein n is an integer with a value of 0 to 50. In certain embodiments, n is an integer with a value of 0 to 10, for example, n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0495] In certain embodiments, the compound is:

[0496]

[0497] In certain embodiments, the compound is 2-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)-4-(6-(piperidin-1-yl)naphthalen-2-yl)nicotinonitrile.

[0498] In certain embodiments, the compound is:

[0499]

[0500] In certain embodiments, the compound is 4-(6-(piperidin-1-yl)naphthalen-2-yl)nicotinonitrile.

[0501] In certain embodiments, the compound is selected from the following:

[0502]

[0503]

[0504]

[0505] or a salt or solvate thereof.

[0506] Treatments and uses

[0507] After a patient has been diagnosed with a traumatic brain injury (TBI), certain procedures may be provided to treat or alleviate the symptoms of the TBI, or to reduce the chance of more damage to the brain. Once a TBI is diagnosed, the progression of the disease may also be monitored by the methods described herein. Once diagnosed, the attending physician may also recommend other treatments as described herein. In one embodiment, once a TBI is detected or suspected, the patient is instructed to avoid active physical activity (e.g., sports, military service).

[0508] In some embodiments, an agent that treats or ameliorates TBI may be administered to a patient. Examples of treatments (agents) for TBI are provided below.

[0509] drug

[0510] Sedation: This can help prevent agitation and excessive muscle activity and help relieve pain. Examples include profanol.

[0511] Pain relief: Opioids may be used.

[0512] Diuretics: These increase urine output and reduce the amount of fluid in tissues. These are given intravenously. Mannitol is the most commonly used diuretic for people with TBI.

[0513] Anti-seizure drugs: People who experience moderate to severe TBI may have seizures for up to a week after the event. Medication can help prevent further brain damage from seizures.

[0514] Coma-inducing drugs: During a coma, a person needs less oxygen. Sometimes, a coma can be intentionally induced if the blood vessels are unable to supply adequate amounts of food and oxygen to the brain.

[0515] Operation

[0516] Surgery may be necessary in some cases.

[0517] Removing a hematoma: Internal bleeding can cause some or all of the clotted blood to accumulate in certain parts of the brain, which can increase pressure on brain tissue. Emergency surgery can remove a hematoma between the skull and brain, thereby reducing pressure inside the skull and preventing further brain damage.

[0518] Repairing a skull fracture: Any part of the skull that is broken and pressing into the brain needs to be surgically repaired. Skull fractures that are not pressing into the brain usually heal on their own. The main problem with a skull fracture is that the force that was enough to cause the skull fracture can cause further potential damage.

[0519] Creating an opening in the skull: This can relieve pressure inside the skull if other interventions have not worked.

[0520] Long-term treatment

[0521] People with severe TBI may need rehabilitation.

[0522] Depending on the extent and type of injury, they may need to relearn how to walk, talk, and perform other daily tasks.

[0523] This may include treatment in a hospital or specialized treatment center. Depending on the type of injury, it may involve physical therapists, occupational therapists, and others.

[0524] "Treatment" or "treatment" is a method of obtaining beneficial or desired results (including clinical results). Beneficial or desired clinical results may include one or more of the following: a) inhibiting a disease or condition (e.g., reducing one or more symptoms caused by the disease or condition, and / or reducing the extent of the disease or condition); b) improving, slowing, or arresting 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) relieving the disease, i.e., causing regression of clinical symptoms (e.g., improving the disease state, partially or completely relieving the disease or condition, enhancing the efficacy of another drug, delaying the progression of the disease, improving the quality of life, and / or prolonging survival).

[0525] "Prevention" or "preventing" refers to the treatment of any disease or condition that results in clinical symptoms of the disease or condition not developing. In some embodiments, the compound can be administered to a subject (including a human) who is at risk for or has a family history of the disease or condition.

[0526] "Subject" refers to an animal, such as a mammal (including a human), that has been or will be the subject of treatment, observation, or experiment. The methods described herein can be used for human treatment and / or veterinary applications. In some embodiments, the subject is a mammal. In one embodiment, the subject is a human.

[0527] The term "therapeutically effective amount" or "effective amount" of a compound described herein, or a pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug, or deuterated analog thereof, refers to an amount sufficient to treat, provide a therapeutic benefit such as symptom improvement or slowed disease progression when administered to a subject. For example, a therapeutically effective amount may be an amount sufficient to alleviate the symptoms of a disease or condition of TBI. The therapeutically effective amount may vary depending on the subject and the disease or condition being treated, the subject's weight and age, the severity of the disease or condition, and the mode of administration, which can be readily determined by one of ordinary skill in the art.

[0528] The methods described herein can be applied to cell populations in vivo or in vitro. "In vivo" means in a living individual, such as in an animal or human. In this article, the methods described herein can be used therapeutically in an individual. "Ex vivo" means outside a living individual. Examples of ex vivo cell populations include in vitro cell cultures and biological samples, which include fluid 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 article, the compounds and compositions described herein can be used for a variety of purposes, including therapeutic and experimental purposes. For example, for a given indication, cell type, individual, and other parameters, the compounds and compositions described herein can be used in vitro to determine the optimal dosing regimen and / or dosage of the compounds of the present invention. The information collected from this use can be used for experimental purposes or in the clinic for setting up an in vivo treatment regimen. Other ex vivo uses for which the compounds and compositions described herein may be suitable are described below, or will become apparent to those skilled in the art. The selected compounds can be further characterized to check the safety or tolerable dose of human or non-human subjects. Such properties can be checked using methods known to those skilled in the art.

[0529] Administration and pharmaceutical compositions

[0530] In some cases, the probe or compound is applied to the eye. In some cases, the pharmaceutical composition of the present disclosure applied to the eye is delivered to the retina, the intraocular space, the ocular surface, interconnected nerves, conjunctiva, lacrimal glands or meibomian glands. In some cases, the compound is topically applied to the eye. In some cases, the compound is administered in the form of eye drops.

[0531] The probes or compounds may also be formulated for intravenous and subcutaneous use without limitation. Intravenous administration may be by bolus administration or continuous infusion.

[0532] The probe is effective in a wide dosage range. In some cases, in the case of adults, 0.01 to 1000 mg, 0.5 to 100 mg, 1 to 50 mg, 5 to 40 mg per day are examples of dosages used. An exemplary dosage is 10 to 30 mg per day. In the dosage for teenagers, the dosage may be equal to or less than the adult dosage. In some cases, the effective amount of the probe is equivalent to about 50-500 mg of the compound per adult subject. The exact dosage will depend on the route of administration, the form of administration of the compound, the subject to be treated, the weight of the subject to be treated, and the preference and experience of the attending physician.

[0533] In some cases, the effective amount of the probe corresponds to about 0.01-1000 mg of the compound per dose per human subject. In some cases, the effective dose of the compound is 50-500 mg per dose per human subject. In some cases, the effective amount corresponds to about 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, 0.1-500 mg, 0.1-600 mg, 0 .1-700mg, 0.1-800mg, 0.1-900mg, 0.1-1000mg, 1-100mg, 1-200mg, 1-300mg, 1-400mg, 1-500mg, 1-600mg, 1-700mg , 1-800mg, 1-900mg, 100-200mg, 100-300mg, 100-400mg, 100-500mg, 100-600mg, 100-700mg, 100-800mg, 100-900mg , 100-1000mg, 200-300mg, 200-400mg, 200-500mg, 200-600mg, 200-700mg, 200-800mg, 200-900mg, 200-1000mg, 30 0-400mg, 300-500mg, 300-600mg, 300-700mg, 300-800mg, 300-900mg, 300-1000mg, 400-500mg, 400-600mg, 400-70 In some embodiments, the present invention relates to an oral dosage form of at least one iodine-containing pharmaceutical composition. The oral dosage form of at least one iodine-containing pharmaceutical composition can be 0 mg, 400-800 mg, 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 about 900-1000 mg. In some cases, the effective amount corresponds to about 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, 300-400 mg, 300-500 mg or 400-500 mg per adult per dose.

[0534] In some cases, the probe is administered in a single dose. In some cases, the probe of the present disclosure is administered in multiple doses. In some cases, the administration is about once, twice, three times, four times, five times, six times or more than six times a day. In some cases, the administration is about once a month, once every two weeks, once a week or once every other day. In another case, the probe and another agent are administered together about once a day to about 6 times a day. In some cases, the administration of the probe and the agent lasts less than about 7 days. In another case, the administration lasts more than about 6, 10, 14, 28 days, two months, six months or one year. In some cases, continuous administration can be achieved and maintained as long as necessary.

[0535] In some cases, the probe is applied 1 to 10 times, 1 to 4 times or once a day. In some cases, the probe is applied 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 times a day. In some cases, the probe is applied in the form of drops. In some cases, the size of the drops applied is about 10-100 μL, about 10-90 μL, about 10-80 μL, about 10-70 μL, about 10-60 μL, about 10-50 μL, about 10-40 μL, about 10-30 μL, about 20-100 μL, about 20-90 μL, about 20-80 μL, about 20-70 μL, about 20-60 μL, about 20-50 μL, about 20-40 μL or about 20-30 μL. An example of the present disclosure applies drops in the range of about 10 to about 30 μL. An example of the present disclosure applies drops in the range of about 10 to about 100 μL. An example of the present disclosure applies drops in the range of about 20 to about 50 μL. An example of the present disclosure applies drops in the range of about 20 to about 40 μL. An example of the present disclosure applies drops in the range of about 10 to about 60 μL. In some cases, the ophthalmic preparation of the present disclosure applies a few drops each time, such as 1-3 drops each time, 1-3 drops each time, 1-4 drops each time, 1-5 drops each time, 1-6 drops each time, 1-7 drops each time, 1-8 drops each time, 1-9 drops each time, 1-10 drops each time, 3-4 drops each time, 3-5 drops each time, 3-6 drops each time, 3-7 drops each time, 3-8 drops each time, 3-9 drops each time, 3-10 drops each time, 5-6 drops each time, 5-7 drops each time, 5-8 drops each time, 5-9 drops each time, 5-10 drops each time, 7-8 drops each time, 7-9 drops each time or 9-10 drops each time. In one example, the formulation of the present disclosure is administered about 1 drop at a time and 1-6 times per day.

[0536] Pharmaceutical compositions / preparations

[0537] In some cases, the probes described herein are formulated into pharmaceutical compositions. In some cases, pharmaceutical compositions are formulated in a conventional manner using one or more physiologically acceptable carriers, including excipients and adjuvants, which aid in processing the active compound into a pharmaceutically acceptable preparation. The correct formulation depends on the selected route of administration. Any pharmaceutically acceptable techniques, carriers, and excipients are used to prepare the pharmaceutical compositions described herein: 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).

[0538] The pharmaceutical compositions provided herein include probes as described herein and a pharmaceutically acceptable diluent, excipient or carrier. In some cases, such as in combination therapy, the compound is administered as a pharmaceutical composition in which one or more probes are mixed with other active ingredients. In certain cases, the pharmaceutical composition includes one or more probes as described herein.

[0539] As used herein, a pharmaceutical composition refers to a mixture of a compound of any probe described herein with other chemical components such as carriers, stabilizers, diluents, dispersants, suspending agents, thickeners and / or excipients. In some cases, a pharmaceutical composition facilitates administration of a compound to an organism. In some cases for practicing the methods of treatment 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 a mammal suffering from a disease or condition to be detected, diagnosed or treated. In certain cases, the mammal is a human being. In some cases, the therapeutically effective amount depends on the severity of the disease, the age and relative health of the subject, the efficacy 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.

[0540] In some cases, one or more probes are formulated in an aqueous solution. In certain cases, by way of example only, the aqueous solution is selected from a physiologically compatible buffer such as Hank's solution, Ringer's solution, acetate buffered aqueous solution, citrate buffered aqueous solution, carbonate buffered aqueous solution, phosphate buffered aqueous solution, or physiological saline buffer.

[0541] In some cases, the compounds described herein are formulated for intraocular administration. In some cases, ophthalmic preparations are liquid (solutions, suspensions, reconstituted powders, sol-to-gel systems), semisolid (ointments and gels), solid (ophthalmic preparations) and intraocular dosage forms (injections, irrigants and implants).

[0542] Provided herein are ophthalmic formulations comprising a compound described herein and an ophthalmologically acceptable component. The ophthalmic formulations can be administered in any form suitable for ophthalmic pharmaceutical administration, for example, as a solution, suspension, ointment, gel, liposomal dispersion, colloidal particle suspension, or the like, or in an ocular insert, for example, in an optionally biodegradable controlled release polymer matrix. Importantly, at least one component of the formulation, and preferably two or more of the formulation components, are "multifunctional" in that they can be used to prevent or treat multiple conditions and diseases, or have more than one mechanism of action, or both.

[0543] A "pharmaceutically acceptable" or "ophthalmologically acceptable" component refers to a component that is not biologically or otherwise undesirable, i.e., the component can be incorporated into the ophthalmic preparation of the present invention and topically applied to the patient's eye without causing any undesirable biological effect or interacting in a harmful manner with any other component contained in the preparation composition. When the term "pharmaceutically acceptable" is used to refer to a component other than a pharmacologically active agent, it means that the component has met the required standards of toxicology and manufacturing testing, or is included in the Inactive Ingredient Guide prepared by the U.S. Food and Drug Administration.

[0544] The formulation also includes an effective amount of a penetration enhancer that promotes the penetration of formulation components through cell membranes, tissues, and extracellular matrices, including the cornea. An "effective amount" of a penetration enhancer means a concentration sufficient to provide a measurable increase in the penetration of one or more formulation components through membranes, tissues, and extracellular matrices, as just described. Suitable penetration enhancers include, for example, methylsulfonylmethane (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, with MSM being particularly preferred.

[0545] Kits and Packaging

[0546] Also provided herein are kits and packages comprising a probe of the invention, a retinal imaging device, and optionally suitable packaging. In one embodiment, the kit further comprises instructions for use.

[0547] The retinal imaging device may include a lens and an image sensor (thereby forming a suitable retinal scanner) for detecting signals emitted from the probe used. In some embodiments, the retinal imaging device detects fluorescent signals. In some embodiments, the retinal imaging device also includes a laser light source, which can be used to activate the fluorescent signal from the probe.

[0548] Example

[0549] The following examples are included to illustrate specific embodiments of the present disclosure. It will be appreciated by those skilled in the art that the techniques disclosed in the following examples represent techniques that function well in the practice of the present disclosure, and therefore can be considered to constitute a specific mode of its practice. However, it will be appreciated by those skilled in the art that, without departing from the spirit and scope of the present disclosure, many changes can be made to the disclosed specific embodiments and the same or similar results can be obtained according to the content disclosed in the present invention.

[0550] Example 1: In vitro analysis of probes and human AD tissues

[0551] Postmortem retinal tissue and brains from humans with confirmed or probable Alzheimer's disease (AD) were obtained and examined by staining for amyloid beta (Aβ) using the test amyloid-sensitive fluorescent probe (ASF) Compound 1. This example was conducted to determine whether ASF could be used to identify patients with amyloid burden in the brain by fluorescent examination of ASF-stained retinas.

[0552] like Figure 1 As shown, dense deposits of 5-10 μm in diameter can be observed fluorescently in flat-mounted retinal samples stained with ASF. Although these deposits can be observed in AD (panels AB) and cognitively normal patients (panels CD), there is a qualitative correlation between the density of these deposits in the retina and the extent of amyloidosis in the brain (as estimated by immunohistochemistry (IHC) with anti-Aβ antibodies).

[0553] IHC staining of retinal cryosections provided further evidence for the presence of Aβ in retinal deposits, which were similar in location, size, and morphology to those found in ASF-stained retinal flat mounts. This provided promising evidence that ASF is able to bind to amyloid in isolated human retinal tissue.

[0554] Example 2: Ocular Testing Using the TBI Blast Mouse Model

[0555] This example was performed to determine whether ASF Compound 1 could detect Aβ in the retina of a TBI mouse model after injury.

[0556] Retinal tissues from blast mice and uninjured mice 24 hours after injury were stained with ASF and co-stained with anti-Aβ antibody (6E10). Figure 2 As shown, mice that had sustained blast injuries showed immunoreactivity to Aβ in their retinal tissue, whereas uninjured mice showed no response to 6E10. More importantly, ASF labeled these retinal deposits with fluorescence ( Figure 2 , top row), but in uninjured mice ( Figure 2 , lower row) did not show any fluorescence enhancement.

[0557] This provides evidence that ASF probe compound 1 has the ability to detect retinal amyloid present in the TBI mouse model. Based on this evidence, this example further attempted to confirm that Aβ was also detected in brain tissue of the same TBI blast mouse model. Figure 3 As shown, mice subjected to blast injury showed immunoreactivity for Aβ in brain tissue, whereas uninjured mice showed no response to 6E10. The evidence for Aβ in brain tissue complements that found in retinal tissue from the same injured mice. It also supports previous studies of a correlation between the brain and retina in Aβ accumulation.

[0558] Example 3: Controlled cortical shock in C5BL / 6 mice

[0559] 3-month-old C5BL / 6 mice were briefly anesthetized with isoflurane and placed in a stereotaxic frame. Mice were craniotomized in the right primary and secondary motor cortex and parietotemporal cortex (+1 to -4 before and after the bregma, 4 mm laterally from the sagittal suture). Half of the mice were not further injured to serve as control samples. For the remaining mice, a 3mm diameter piston was placed centered above the motor cortex, approximately +0.5 to -2.5mm from the bregma, and 3mm laterally from the sagittal suture. Using a stereotaxic impactor (Impact One, myNeuroLab.com), the piston was accelerated to an impact depth of 1mm below the cortical surface at a speed of 3m / sec. After 24 hours, the mice were euthanized and the brains and eyes were collected. The retina was extracted, stained with DAPI, compound 1, and 6E10 antibodies, and then compared.

[0560] The retinas were mounted on slides and washed twice with phosphate buffered saline (PBS) for five minutes each. The retinas were exposed to 98% formic acid for one minute and then washed twice with distilled water for five minutes each. The retinas were then equilibrated in 1xPBS for 15 minutes.

[0561] 20 (PBST) in 1x phosphate buffered saline with 10% goat / donkey serum for 1 hour. The retina was then incubated with 610 antibody in 10% goat serum in 1x PBST at 4°C overnight. The retina was then rinsed 3 times with 1x PBST for 5 minutes each. The retina was then incubated with the secondary antibody Alexa Fluor 568 anti-mouse in 1x PBST for 1 hour while protecting from light. The retina was rinsed 3 times with 1x PBST for 5 minutes each. The retina was then stained with compound 1 working solution for 30 minutes at room temperature while protecting from light. The retina was then washed 3 times with 1x PBS for 5 minutes each and then stained with 300nM or 100ng / mL of DAPI for 10 minutes in the dark. The retina was then washed 3 times with PBS for 10 minutes each. DAKO mounting medium was applied and the samples were stored in the dark until imaging.

[0562] Fluorescence imaging was performed on a Leica DMI 4000B microscope equipped with a TCS SPE camera and Leica 10x, 20x, and 40x objectives. The following lasers were used to visualize fluorescent probes associated with DAPI (blue), compound 1 (green), 6E10 antibody (red): 408, 488, and 568 nm. Z-stack images were taken at 40x in 0.5 μm increments to visualize the entire thickness of the tissue.

[0563] Figure 4-7 Demonstrated that after CCI, 3-month-old C5BL / 6 mice developed deposits of aggregated proteins, which were detected in the retina by both compound 1 and 6E10 antibodies. Aggregated proteins were detected in the retina by 6E10 and co-localized by compound 1.

[0564] Example 4: Detection of retinal Aβ aggregation in an ex vivo mouse model

[0565] This example was performed to examine whether TBI induces retinal Aβ aggregation in a mouse model.

[0566] Mice were subjected to controlled cortical impact (CCI). The CCI model applies a controlled impact to the intact dura mater after craniotomy and is a commonly used animal model of TBI. Mice were anesthetized, the head was mounted in a stereotaxic frame, and a craniotomy was performed on the right side of the frontal cortex. Using a stereotactic impactor, mice were impacted on the right side of the frontal cortex with a piston at a speed of 3m / s, a depth of 1mm, and a diameter of 3mm. After injury, the incision was closed with staples, anesthesia was terminated, and the animals were placed in a heated cage to maintain normal core temperature. Sham injury involves exposure to anesthesia, stereotactic mounting, skin and fascia reflexes, and closure of the incision with staples. The mice were then euthanized 24 hours after injury, and retinal tissue was dissected for further analysis.

[0567] The mouse retinas were extracted and stained with compound 1 and 6E10 antibody (sequence-specific antibody for Aβ). Figure 8 As shown, mice that underwent CCI showed immunoreactivity for Aβ along blood vessels in the retina, whereas uninjured mice showed no response to 6E10. More importantly, compound 1 fluorescently labeled these deposits and colocalized with 6E10 immunoreactivity ( Figure 8 Upper row, white arrows), but did not show any fluorescence enhancement in uninjured mice ( Figure 8 This example demonstrates that the compounds of the present disclosure can effectively detect retinal amyloid in a TBI mouse model.

[0568] Example 4: Detection of retinal Aβ aggregation in an ex vivo mouse model

[0569] This example was performed to examine whether TBI induces retinal Aβ aggregation in a mouse model.

[0570] Mice were subjected to controlled cortical impact (CCI). The CCI model applies a controlled impact to the intact dura mater after craniotomy and is a commonly used animal model of TBI. Mice were anesthetized, the head was mounted in a stereotaxic frame, and a craniotomy was performed on the right side of the frontal cortex. Using a stereotactic impactor, mice were impacted on the right side of the frontal cortex with a piston at a speed of 3m / s, a depth of 1mm, and a diameter of 3mm. After injury, the incision was closed with staples, anesthesia was terminated, and the animals were placed in a heated cage to maintain normal core temperature. Sham injury involves exposure to anesthesia, stereotactic mounting, skin and fascia reflexes, and closure of the incision with staples. The mice were then euthanized 24 hours after injury, and retinal tissue was dissected for further analysis.

[0571] The mouse retinas were extracted and stained with compound 1 and 6E10 antibody (sequence-specific antibody for Aβ). Figure 8As shown, mice that underwent CCI showed immunoreactivity for Aβ along blood vessels in the retina, whereas uninjured mice showed no response to 6E10. More importantly, compound 1 fluorescently labeled these deposits and colocalized with 6E10 immunoreactivity ( Figure 8 Upper row, white arrows), but did not show any fluorescence enhancement in uninjured mice ( Figure 8 This example demonstrates that the compounds of the present disclosure can effectively detect retinal amyloid in a TBI mouse model.

[0572] Example 5: Synthesis

[0573] Synthesis of compound 21

[0574]

[0575] Synthesis of (6-bromonaphthalen-2-yl)methanol (25)

[0576] Under N2, at 0 ℃, to LiAlH4(8.2g, 217mmol in 500mL THF) solution) dropwise add 6-bromo-2-naphthoic acid methyl ester (24) (50.0g, 189mmol) in 500mL anhydrous THF solution.Reaction mixture was stirred at 0 ℃ for 1 hour.After completing the reaction by TLC monitoring, the mixture was treated with H2O, 15%NaOH, H2O (1: 1: 3, v / v / v).After filtering, the filtrate was concentrated and extracted with EA, through NaSO4 drying.From (PE: EA=3: 1) purification of crude product, obtain title compound.

[0577] Synthesis of 6-bromo-2-naphthaldehyde (26)

[0578] In a suspension of (6-bromonaphthalene-2-yl)methanol (25) (42.0 g, 177 mmol) and silica gel (76.4 g) in DCM (500 mL), PCC (76.4 g, 354 mmol) was added. The reaction was stirred at RT (room temperature, 25 ± 5 ° C) for 1.5 hours. After completion, it was filtered through a silica pad and concentrated under reduced pressure to obtain the title compound.

[0579] Synthesis of 6-(piperidin-1-yl)-2-naphthaldehyde (27)

[0580] In dry 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 was stirred at 115°C for 8 hours. After cooling, the mixture was filtered and washed with EA, then concentrated to one third of the volume, to which 200 ml of 6N hydrochloric acid was added with sufficient stirring. The aqueous phase was separated and extracted three times with DCM, then adjusted to alkalinity with 5N NaOH and extracted with EA. The organic phase was concentrated to give a crude product, which was further purified by silica gel chromatography (PE:EA=20:1 to 2:1) to give the title compound.

[0581] Synthesis of 2-cyano-N-(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)acetamide (29)

[0582] In a pear-shaped flask, 28 (6.0 g, 40 mmol) was added to methyl 2-cyanoacetate (4.0 g, 40 mmol) under stirring. The mixture was stirred at room temperature overnight and concentrated to a crude material. The crude material was used directly in the next step.

[0583] Synthesis of (E)-2-cyano-N-(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)-3-(6-(piperidin-1-yl)naphthalen-2-yl)acrylamide (Compound 21)

[0584] To a solution of 6-(piperidin-1-yl)-2-naphthaldehyde (27) (7.0 g, 29.3 mmol) and 2-cyano-N-(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)acetamide (29) (7.9 g, 36.5 mmol) in anhydrous THF (250 mL) was added piperidine (0.5 g, 5.9 mmol), and the resulting mixture was refluxed for 12 hours. The reaction mixture was then concentrated under reduced pressure to a crude material, 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%, O14.63%.

[0585] Synthesis of compound 22

[0586]

[0587] Synthesis of 31

[0588] To 1 g (2.3 mM) of compound 21 in 15 mL of THF at 0 degrees was added 1.6 g (3 mM) of tetrabenzyl pyrophosphate prepared using the Merck organic synthesis method to give a dark red solution. Sodium hydride (100 mg, 2.5 mM, 60% in oil) was added. After 15 minutes, warm to room temperature and solids began to precipitate. DMF (5 mL) was added and 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 residue was purified by centrifugation on an 80 g silica gel column using 0-100% hexane / ethyl acetate. Purification gave the title compound. 1 H NMR spectrum is shown in Figure 1 A to 1C. MS (m / z) 701.3 [M+H] + .

[0589] Synthesis of compound 22

[0590] In 1 gram (compound 31), add 95% ethanol (150mL) degassed with argon. Add 120mg of 10% Pd / C, and blast hydrogen into the reaction mixture for 5 minutes. Then the mixture was stirred for 3 hours under a hydrogen balloon. The reaction mixture was degassed with argon and evaporated. Purification was carried out by using 0 to 100% water (containing 2g per liter of ammonium acetate) on a C18 chromatographic column of 25x250 mm. After free drying, compound 22 was obtained.

[0591] Synthesis of compound 23

[0592]

[0593] A solution of compound 31 (5 mmol, 3.49 g, 1 eq.) in anhydrous CH2Cl2 (100 ml) was cooled to 0°C under argon and trimethylsilyl bromide (50 mmol, 6.8 ml, 10 eq.) was added via 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.

[0594] The organic solvent was evaporated in vacuo, the residue was suspended in a trace amount of MeOH, and EtOAc was added to induce precipitation of phosphonic acid. The residue was filtered and washed with EtOAc (x2). The residue was then dried in vacuo to obtain the desired phosphoric acid.

[0595] The phosphonic acid was treated with NH4OAc (25 mmol, 1.93 g, 5 eq) and 150 ml of water at room temperature and stirred for another 15 minutes 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] + . 1H-NMR: (400MHz, CD3OD) δ8.31–8.22(m,2H),8.08(dd,J=8.8,1.9Hz,1H),7.81(d,J=9.2Hz,1H),7.74(d,J=8.8Hz,1H),7.40(dd,J=9.2 ,2.5Hz,1H),7.18(d,J=2.5Hz,1H),4.08–3.98(m,2H),3.79–3.65(m,8H),3.58(t,J=5.5Hz,2H),3.45–3.39(m,4H),1.86–1.63(m,6H).

[0596] Example 6: In vivo detection of retinal Aβ aggregation

[0597] Following the positive ex vivo experiments in Example 4, in vivo retinal imaging studies were performed using mice before and after receiving CCI.

[0598] Mice were imaged prior to TBI via CCI to obtain baseline retinal images. Anesthetized mice received intravenous administration of compound 23 (15 mg / kg in 20 mg / mL in 0.1 M phosphate buffer, pH 7.4) prior to CCI and 24 hours after CCI. Fig. 9 As shown, an increase in background fluorescence was observed in the retinal vasculature 3 minutes after administration of compound 23 to uninjured mice. This increase was no longer observed 15 minutes after administration of compound 23 ( Fig. 9 , top row).

[0599] 24 hours after baseline imaging, mice were anesthetized, their heads fixed in a stereotaxic frame, and a craniotomy was performed on the right side of the motor cortex. Using a stereotaxic impactor, mice were impacted with a piston at a speed of 5 m / s, a depth of 2 mm, and a diameter of 3 mm on the right side of the motor cortex. 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. 24 hours after CCI, real-time retinal imaging was performed on anesthetized mice after intravenous injection of compound 23. As Fig. 9 As shown, the retinal vasculature remained illuminated during the 15-minute imaging period, indicating that Aβ can persist in the vessels ( Fig. 9 , bottom row). This is in contrast to pre-CCI imaging, which showed increased fluorescence in the vessels 3 minutes after injection but decreased over the 15-minute imaging period.

[0600] This example demonstrates that compound 23 can be used to detect changes in the retinal vasculature that may indicate accumulation of Aβ within 24 hours after CCI. Figure 8 This is consistent with the ex vivo data shown, where amyloid accumulation was observed along the blood vessels of mice 24 hours after CCI.

[0601] ***

[0602] Unless defined otherwise, 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 invention belongs.

[0603] The invention illustratively described herein may be suitably practiced in the absence of any element or elements, limitation or limitations not specifically disclosed herein. Thus, for example, the terms "comprises," "comprising," "containing," and the like should be interpreted broadly and without limitation. Furthermore, the terms and expressions employed herein have been used as terms of description rather than limitation, and in the use of said terms and expressions it is not intended to exclude any equivalents of the features shown and described, or portions thereof, but it should be recognized that various modifications within the scope of the claimed invention are possible.

[0604] Therefore, it should be understood that, although the present invention has been specifically disclosed by preferred embodiments and optional features, those skilled in the art may modify, improve and vary the invention specifically disclosed herein, and these modifications, improvements and variations are considered to be within the scope of the present invention. The materials, methods and examples provided herein represent preferred embodiments, are exemplary, and are not intended to limit the scope of the present invention.

[0605] The invention has been described broadly and generally herein. Each of the narrower species and subgeneric groupings falling within the generic disclosure also form part of the invention. This includes the generic description of the invention with proviso or negative limitation removing any subject matter from the genus, whether or not the removed material is specifically recited herein.

[0606] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0607] All publications, patent applications, patents, and other references mentioned herein are expressly incorporated by reference in their entirety to the same extent as if each were individually incorporated by reference. In the event of conflict, the present specification, including definitions, will control.

[0608] It should be understood that although the present disclosure has been described in conjunction with the above embodiments, the foregoing description and examples are intended to illustrate rather than limit the scope of the present disclosure. For those skilled in the art to which the present disclosure belongs, other aspects, advantages and modifications within the scope of the present disclosure will be apparent.

Claims

1. A method of determining whether a patient has suffered a traumatic brain injury (TBI), comprising detecting the presence of amyloid beta protein in an eye of the patient.

2. The method of claim 1, wherein the detection is for amyloid beta protein in the retina of the eye.

3. The method according to claim 1 or 2, wherein the patient has received a physical impact to the head within 30 days prior to the detection.

4. The method of claim 3, wherein the patient has received a physical impact to the head within 24 hours prior to the testing.

5. A method according to any preceding claim, wherein the patient is not subjected to direct physical or light impact on the eye.

6. A method according to any preceding claim, wherein the patient is not known or suspected to have Alzheimer's disease.

7. The method according to any preceding claim, wherein the patient is a human under 40 years of age.

8. The method of any preceding claim, wherein the detecting comprises contacting the amyloid-β protein with a probe in vivo.

9. The method of claim 8, wherein the contacting causes emission of a detectable signal when activated by light.

10. The method of claim 9, wherein the detectable signal is a fluorescent or infrared signal.

11. The method according to any one of claims 8 to 10, wherein the probe comprises a compound of formula Ic: in EDG is: a) a heterocycloalkyl group of not more than 10 carbon atoms, which is optionally substituted by one or more R 17 replace; or b)-NR 10 R 11 ; Each R 17 are independently halogen, -OR 18 、-NR 19 R 20 、C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons; R 10 , R 11 , R 18 , R 19 and R 20 Each of which is independently hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbon atoms, heterocycloalkyl of up to 10 carbon atoms, arylene of up to 10 carbon atoms, or heteroarylene of up to 10 carbon atoms, each of which is optionally replaced by one or more R 21 replace; R 21 Each of which is independently halogen, -OR 22 、-NR 23 R 24 、C1-C 10 Alkyl, C1-C 10 is a heteroalkyl, a cycloalkyl of up to 10 carbon atoms, a heterocycloalkyl of up to 10 carbon atoms, an arylene of up to 10 carbon atoms, or a heteroarylene of up to 10 carbon atoms, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally replaced by one or more R 25 replace; R 22 , R 23 and R 24 Each of 10 Alkyl; and Each R 25 Independently C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons; Ar is an arylene group of up to 14 carbon atoms or a heteroarylene group of up to 14 carbon atoms, each optionally substituted with one or more R1; Each of R1 is independently halogen, -OR2, -NR3R4, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted with one or more R5; R2, R3 and R4 are independently hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons, each of which is optionally substituted with one or more R5, excluding hydrogen; Each R5 is independently halogen, -OR6, -NR7R8, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons; R6, R7, R8 and R 84 are independently hydrogen or C1-C 10 alkyl; EWG is selected from -F, -Cl, -Br, -CH=O, NO2, -CF3, -CCl3, -SO3H and -CN; WSG is: i) ii) polyethylene glycol, polypropylene glycol, copolymers of polyethylene glycol and polypropylene glycol or alkoxy derivatives thereof; iii) Where n is an integer from 1 to 50 and R 81 is hydrogen, C1-C 10 Alkyl, C1-C 10 Alkenyl or C1-C 10 Alkynyl, wherein the alkyl, alkenyl or alkynyl group is optionally substituted with one or more C1-C 10 Alkyl, C1-C 10 substituted with heteroalkyl, cycloalkyl of up to 10 carbon atoms, heterocycloalkyl of up to 10 carbon atoms, arylene of up to 10 carbon atoms, or heteroarylene of up to 10 carbon atoms; iv) v) vi) -(C1-C 10 Alkyl)-R 33 -R 37 ,in: R 33 is a heteroarylene group having no more than 10 carbon atoms; and R 37 is -(C1-C6 alkyl)(heterocycloalkyl of not more than 10 carbons); vii) viii) –(C1-C 10 Heteroalkyl)-R 33 -R 37 ,in: R 33 is a heteroarylene group having no more than 10 carbon atoms; and R 37 is -(C1-C6 alkyl)(heterocycloalkyl of not more than 10 carbons); or ix) X is C=O or SO2, or X and R 84 Combine to form pyridyl; Y is NH or S.

12. The method of any one of claims 8-10, wherein the probe comprises an antibody.

13. The method of any preceding claim, further comprising determining that the patient has TBI if amyloid beta is detected in the eye.

14. The method of claim 13, further comprising instructing the patient to refrain from active physical activity.

15. The method of claim 13, further comprising administering to the patient an agent that treats or ameliorates TBI.

16. A method of preparing a patient for diagnosis of traumatic brain injury (TBI) comprising administering to an eye of the patient a probe that specifically binds to amyloid beta protein.

17. The method of claim 16, further comprising detecting binding of the probe to amyloid-β protein in the eye.

18. The method of claim 16 or 17, wherein the binding causes emission of a detectable signal when activated by light.

19. The method of claim 18, wherein the signal is a fluorescent or infrared signal.

20. The method of any one of claims 16-19, wherein the probe comprises a compound of formula Ic: in EDG is: a) a heterocycloalkyl group of not more than 10 carbon atoms, which is optionally substituted by one or more R 17 replace; or b)-NR 10 R 11 ; Each R 17 are independently halogen, -OR 18 、-NR 19 R 20 、C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons; R 10 , R 11 , R 18 , R 19 and R 20 Each of which is independently hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbon atoms, heterocycloalkyl of up to 10 carbon atoms, arylene of up to 10 carbon atoms, or heteroarylene of up to 10 carbon atoms, each of which is optionally replaced by one or more R 21 replace; R 21 Each of which is independently halogen, -OR 22 、-NR 23 R 24 、C1-C 10 Alkyl, C1-C 10 is a heteroalkyl, a cycloalkyl of up to 10 carbon atoms, a heterocycloalkyl of up to 10 carbon atoms, an arylene of up to 10 carbon atoms, or a heteroarylene of up to 10 carbon atoms, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally replaced by one or more R 25 replace; R 22 , R 23 and R 24 Each of 10 Alkyl; and Each R 25 Independently C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons; Ar is an arylene group of up to 14 carbon atoms or a heteroarylene group of up to 14 carbon atoms, each optionally substituted with one or more R1; Each of R1 is independently halogen, -OR2, -NR3R4, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted with one or more R5; R2, R3 and R4 are independently hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons, each of which is optionally substituted with one or more R5, excluding hydrogen; Each R5 is independently halogen, -OR6, -NR7R8, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons; R6, R7, R8 and R 84 are independently hydrogen or C1-C 10 alkyl; EWG is selected from -F, -Cl, -Br, -CH=O, NO2, -CF3, -CCl3, -SO3H and -CN; WSG is: i) ii) polyethylene glycol, polypropylene glycol, copolymers of polyethylene glycol and polypropylene glycol or alkoxy derivatives thereof; iii) Where n is an integer from 1 to 50 and R 81 is hydrogen, C1-C 10 Alkyl, C1-C 10 Alkenyl or C1-C 10 Alkynyl, wherein the alkyl, alkenyl or alkynyl group is optionally substituted with one or more C1-C 10 Alkyl, C1-C 10 substituted with heteroalkyl, cycloalkyl of up to 10 carbon atoms, heterocycloalkyl of up to 10 carbon atoms, arylene of up to 10 carbon atoms, or heteroarylene of up to 10 carbon atoms; iv) v) vi) -(C1-C 10 Alkyl)-R 33 -R 37 ,in: R 33 is a heteroarylene group having no more than 10 carbon atoms; and R 37 is -(C1-C6 alkyl)(heterocycloalkyl of not more than 10 carbons); viii) –(C1-C 10 Heteroalkyl)-R 33 -R 37 ,in: R 33 is a heteroarylene group having no more than 10 carbon atoms; and R 37 is -(C1-C6 alkyl)(heterocycloalkyl of not more than 10 carbons); or ix) X is C=O or SO2, or X and R 84 Combine to form pyridyl; Y is NH or S.

21. The method of any one of claims 16-20, wherein the administration is intravenous or into the retina of the eye.

22. The method of any one of claims 16-21, wherein the patient has been subjected to a physical impact to the head within 30 days prior to the testing.

23. The method of any one of claims 16-22, wherein the patient is not known or suspected to have Alzheimer's disease.

24. The method of any one of claims 16-23, wherein the patient is not subjected to direct physical or light impact on the eye.

25. A kit or package comprising a probe that specifically binds to amyloid beta protein and a retinal imaging device.

26. A kit or package according to claim 25, wherein the retinal imaging device comprises a laser light source.

27. The kit or package of claim 25, wherein the retinal imaging device further comprises a retinal scanner.

28. The kit or package of any one of claims 25-27, wherein the probe comprises a compound of formula Ic: in EDG is: a) a heterocycloalkyl group of not more than 10 carbon atoms, which is optionally substituted by one or more R 17 replace; or b)-NR 10 R 11 ; Each R 17 are independently halogen, -OR 18 、-NR 19 R 20 、C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons; R 10 , R 11 , R 18 , R 19 and R 20 Each of which is independently hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbon atoms, heterocycloalkyl of up to 10 carbon atoms, arylene of up to 10 carbon atoms, or heteroarylene of up to 10 carbon atoms, each of which is optionally replaced by one or more R 21 replace; R 21 Each of which is independently halogen, -OR 22 、-NR 23 R 24 、C1-C 10 Alkyl, C1-C 10 is a heteroalkyl, a cycloalkyl of up to 10 carbon atoms, a heterocycloalkyl of up to 10 carbon atoms, an arylene of up to 10 carbon atoms, or a heteroarylene of up to 10 carbon atoms, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally replaced by one or more R 25 replace; R 22 , R 23 and R 24 Each of 10 Alkyl; and Each R 25 Independently C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons; Ar is an arylene group of up to 14 carbon atoms or a heteroarylene group of up to 14 carbon atoms, each optionally substituted with one or more R1; Each of R1 is independently halogen, -OR2, -NR3R4, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene or heteroarylene is optionally substituted with one or more R5; R2, R3 and R4 are independently hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons, each of which is optionally substituted with one or more R5, excluding hydrogen; Each R5 is independently halogen, -OR6, -NR7R8, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of up to 10 carbons, heterocycloalkyl of up to 10 carbons, arylene of up to 10 carbons, or heteroarylene of up to 10 carbons; R6, R7, R8 and R 84 are independently hydrogen or C1-C 10 alkyl; EWG is selected from -F, -Cl, -Br, -CH=O, NO2, -CF3, -CCl3, -SO3H and -CN; WSG is: i) ii) polyethylene glycol, polypropylene glycol, copolymers of polyethylene glycol and polypropylene glycol or alkoxy derivatives thereof; iii) Where n is an integer from 1 to 50 and R 81 is hydrogen, C1-C 10 Alkyl, C1-C 10 Alkenyl or C1-C 10 Alkynyl, wherein the alkyl, alkenyl or alkynyl group is optionally substituted with one or more C1-C 10 Alkyl, C1-C 10 substituted with heteroalkyl, cycloalkyl of up to 10 carbon atoms, heterocycloalkyl of up to 10 carbon atoms, arylene of up to 10 carbon atoms, or heteroarylene of up to 10 carbon atoms; iv) v) vi) -(C1-C 10 Alkyl)-R 33 -R 37 ,in: R 33 is a heteroarylene group having no more than 10 carbon atoms; and R 37 is -(C1-C6 alkyl)(heterocycloalkyl of not more than 10 carbons); vii) viii) –(C1-C 10 Heteroalkyl)-R 33 -R 37 ,in: R 33 is a heteroarylene group having no more than 10 carbon atoms; and R 37 is -(C1-C6 alkyl)(heterocycloalkyl of not more than 10 carbons); or ix) X is C=O or SO2 or X and R 84 Combine to form pyridyl; Y is NH or S.

29. The method of any one of claims 8-10, 13-19 or 21-24, wherein the probe comprises a compound selected from the group consisting of 30. A kit or package according to any one of claims 25-27, wherein the probe comprises a compound selected from the group consisting of

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