Compositions and methods for detecting traumatic brain injury

Abstract

To provide compositions and methods for determining whether a patient suffers from a traumatic brain injury (TBI) by detecting presence of an amyloid beta protein in an eye of the patient.SOLUTION: Compositions and methods for preparing a patient for diagnosis and treatment of traumatic brain injury (TBI) are also provided. The present disclosure, in some embodiments, provides compositions and methods capable of diagnosing traumatic brain injury (TBI) to reduce a risk of more permanent brain damages. Such methods can be quick and non-invasive. In one example, a non-invasive fluorescent diagnostic probe capable of detecting misfolded protein amyloid-beta (Ap) can be used in a simple ophthalmic examination to detect accumulation of the Aβ in the patient's retina. Such detection which is discovered by the present inventors and can be performed with retinal imaging performs fast and reliable diagnosis of TBI.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 62 / 678,900, filed May 31, 2018, and No. 62 / 733,025, filed September 18, 2018, the contents of each of which are incorporated by reference in their entirety into this application. [Background technology]

[0002] Traumatic brain injury (TBI) is a chronic condition defined as an injury to the brain caused by an external force such as a crash, blow, impact, rapid acceleration or deceleration, or projectile penetration. The injury that leads to TBI can cause a reduced or altered state of consciousness, resulting in temporary or permanent deficits in cognitive, sensorimotor, and psychosocial functioning. Recent studies have shown that nearly 80% of military personnel who have experienced both blast- and non-blast-related mild to severe TBI suffer from moderate to severe global impairment within one year after the injury. Furthermore, several studies have shown that mild TBI is associated with post-traumatic stress disorder (PTSD), depression, and other psychiatric 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 recognized as a major risk factor for the subsequent development of age-related neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), and chronic traumatic encephalopathy (CTE). Recent studies of veterans suggest that the overall risk of PD increases by 71% after any TBI, and even mild TBI may increase the risk by 56%. Veterans are also more likely to develop PD at least 2 years earlier than veterans who develop PD without a TBI.

[0003] The disease can be classified as mild, moderate, or severe, but identifying mild and severe TBI has been problematic because doctors often only assess severity by reference to existing symptoms and subjective measures. Diagnosis of mild TBI, the majority of all brain injuries, is often missed by doctors because the presenting symptoms are similar to those of mental health disorders such as bipolar disorder or depression. As such, mild TBI tends to be underreported and untreated. The Concussion Legacy Foundation estimates that only one in six mild TBIs (also known as concussions) is diagnosed.

[0004] Diagnosis can be further complicated by inconsistent diagnostic criteria that rely heavily on patient self-reported symptoms rather than objective testing. The only way to definitively diagnose TBI is through clinical evaluation and postmortem examination of the brain using standard neuroimaging techniques.

[0005] However, it is now generally established that misfolded protein amyloid beta (Aβ) plaques are found within hours or days after a single TBI, similar to the characteristic Aβ plaque pathology of AD. Studies in humans and pigs immediately after TBI have shown prolonged accumulation of Aβ in injured axons, as well as other proteins involved in the production of Aβ peptides. Recent studies have found Aβ aggregates in the brains of up to one-third of patients who died shortly after TBI, as well as in those who survived for more than a year. Furthermore, approximately one-third of American military personnel who suffered brain injury from a bomb blast showed immediate evidence of injured nerve fibers in the brain, also known as diffuse axonal injury (DAI). Even mild TBI (mTBI) can have profound effects, inducing DAI and causing physical injury and dysfunction of damaged axons. This can result in repetitive mild TBI (rmTBI)—involving multiple mild TBIs—which can exacerbate the injured individual's response and induce Aβ and tau pathology. This evidence suggests that Aβ may be an acute biomarker for diagnosing TBI.

[0006] The current standard diagnosis of severe TBI, or TBI affecting infants and adults over 60 years of age, requires invasive and costly procedures such as intracranial pressure monitoring (ICP) or MRI. Diagnosis of mild TBI is even more complicated because fewer than 10% of patients with mild head trauma have positive findings on computed tomography (CT). Furthermore, the majority of TBI diagnoses are self-report tests, which are subjective and easily manipulated. Self-report tests are particularly problematic because some soldiers may be reluctant to diagnose and manipulate test results to avoid the stigma of being injured or separated from their platoon. Furthermore, the Department of Defense found that 60% of soldiers suffering from TBI symptoms refused assistance because they worried they would be treated differently or because they worried their condition would prevent them from obtaining jobs as police officers and firefighters after they were suspended from duty.

[0007] A very recent 2018 study reported a significantly longer recovery period for women than men after mild TBI. Symptom severity was also higher in women who did not use hormonal contraception (HC) as opposed to those who used HC, and this difference was associated with lower subjective ratings of symptom severity in women who used HC. Because female military personnel make up a significant portion of the military and enlist at the age of reproductive age, the impacts of TBI are even more detrimental. There remains a major clinical need for an objective diagnosis of TBI. Summary of the Invention [Means for solving the problem]

[0008] The present disclosure provides, in some embodiments, compositions and methods that can diagnose 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 that can detect the misfolded protein amyloid beta (Aβ) can be used in a simple ophthalmic examination to detect the accumulation of Aβ in the retina of a patient. Such detection, which the inventors have discovered can be performed with retinal imaging, makes a rapid and reliable diagnosis of TBI.

[0009] 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 that may be used in conjunction with an indirect ophthalmoscope or a smartphone to capture real-time retinal images at the point-of-care.

[0010] The present disclosure provides, in one embodiment, a method for determining whether a patient suffers from traumatic brain injury (TBI). The method can 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.

[0011] In some embodiments, the patient suffered a physical blow to the head within 30, 25, 20, 15, 10, 5, or 2 days prior to detection. In some embodiments, the physical blow was more than 1, 2, 3, 4, 5, 10, 15, 20, 25, or 30 days prior to detection. In some embodiments, the patient suffered a physical blow to the head within 24 hours prior to detection.

[0012] In some embodiments, the patient has not experienced a direct physical or optical impact on the eye. In some embodiments, the patient is not known or suspected to have Alzheimer's disease. In some embodiments, the patient is a human under the age of 40.

[0013] In some embodiments, the detection comprises contacting the amyloid beta protein with the probe in vivo. In some embodiments, the contact causes the 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.

[0014] In some embodiments, the detection comprises imaging the amyloid beta protein with the probe ex vivo. In some embodiments, the contact between the amyloid beta protein and the probe is performed in vivo, and the detection and / or imaging is performed ex vivo. In some embodiments, the sample containing the amyloid beta protein is removed from the subject before the detection and / or imaging.

[0015] In some embodiments, the probe comprises a compound of formula Ic: [ka] During the ceremony, EDG is, a) optionally, one or more R 17 or b) -NR 10 R 11 and In the formula, each R 17 are independently halogen, -OR 18 , -NR 19 R 20 , C1~C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons; R 10 , R 11 , R 18 , R 19 and R20 Each of the groups is independently hydrogen, C1 to C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less, each of which, other than hydrogen, is optionally represented by one or more R 21 is replaced by R 21 Each of the groups is independently a halogen, -OR 22 , -NR 23 R 24 , C1~C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene is optionally further represented by one or more R 25 is replaced by R 22 , R 23 and R 24 Each of is independently hydrogen or C1 to C 10 is alkyl, R 25 Each of C1 to C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons; Ar is an arylene of up to 14 carbon atoms or a heteroarylene of up to 14 carbon atoms, each optionally substituted with one or more R1, each R1 independently being halogen, -OR2, -NR3R4, C1-C 10 Alkyl, C1-C 10heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene is optionally substituted with one or more R; R2, R3, and R4 are independently hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less, each of which, other than hydrogen, is optionally substituted with one or more R; Each R5 is independently halogen, -OR6, -NR7R8, C1 to C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons; R6, R7, R8 and R 84 are independently hydrogen or C1-C 10 is alkyl, EWG is selected from the group consisting of -F, -Cl, -Br, -CH=O, NO2, -CF3, -CCl3, -SO3H and -CN; WSG is i) [ka] or ii) polyethylene glycol, polypropylene glycol, copolymers of polyethylene glycol and polypropylene glycol, or alkoxy derivatives thereof; iii) [ka] (In the formula, n is an integer of 1 to 50, and R 81is hydrogen, C1-C 10 Alkyl, C1-C 10 Alkenyl, or C1-C 10 Alkynyl, where the alkyl, alkenyl, or alkynyl is optionally one or more C1-C 10 Alkyl, C1-C 10 substituted with heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less; iv) [ka] or v) [ka] or vi) -(C1~C 10 Alkyl)-R 33 -R 37 (In the formula, R 33 is a heteroarylene of 10 carbons or less, R 37 is -(C1-C6 alkyl)(heterocycloalkyl of 10 or less carbons) vii) [ka] or viii) -(C1~C 10 Heteroalkyl)-R 33 -R 37 (In the formula, R 33 is a heteroarylene of 10 carbons or less, R 37 is -(C1-C6 alkyl)(heterocycloalkyl of 10 or less carbons); or ix) [ka] and X is C=O or SO2, or X and R 84 and combine to form pyridinyl, Y is NH or S.

[0016] In some embodiments, the probe comprises an antibody. In some embodiments, the antibody is specific to the protein amyloid beta (Aβ). In some embodiments, the antibody selectively binds to the misfolded protein amyloid beta (Aβ). The antibody can be labeled with a detectable marker or can be detected by binding to a molecule associated with the detectable marker.

[0017] In some embodiments, the method further comprises determining that the patient suffers from TBI if amyloid beta protein is detected in the eye. In some embodiments, the method further comprises instructing the patient to refrain from vigorous physical activity. In some embodiments, the method further comprises administering to the patient an agent that treats or ameliorates TBI.

[0018] Also provided in one embodiment is a method for preparing a patient for diagnosis of traumatic brain injury (TBI), comprising administering a probe that specifically binds to amyloid beta protein to the eye of the patient.In some embodiments, the method further comprises detecting the binding of the probe to amyloid beta protein in the eye.Examples of the probe are as provided above.

[0019] 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 eye of a patient, and detecting the binding of the probe to amyloid beta protein ex vivo.In some embodiments, the contact of the amyloid beta protein with the probe is performed in vivo, and the detection is performed ex vivo.In some embodiments, a sample comprising the amyloid beta protein and the probe is removed from the subject prior to detection.

[0020] In some embodiments, the administration is intravenous or localized to the retina of the eye. In some embodiments, the patient has received a physical blow to the head within 30, 25, 20, 15, 10, 5, or 2 days prior to detection. In some embodiments, the physical blow is more than 1, 2, 3, 4, 5, 10, 15, 20, 25, or 30 days prior to detection. In some embodiments, the patient has received a physical blow to the head within 24 hours prior to detection.

[0021] In some embodiments, the patient has not experienced a direct physical or optical impact on the eye. In some embodiments, the patient is not known or suspected to have Alzheimer's disease. In some embodiments, the patient is a human under the age of 40.

[0022] Also provided are kits and packages that include 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. In an embodiment of the present invention, for example, the following items are provided: (Item 1) 1. A method for determining whether a patient has suffered a traumatic brain injury (TBI), comprising detecting the presence of amyloid beta protein in the eye of the patient. (Item 2) 2. The method of claim 1, wherein the detection is for amyloid beta protein in the retina of the eye. (Item 3) 3. The method of claim 1 or 2, wherein the patient has suffered a physical blow to the head within 30 days prior to said detection. (Item 4) 4. The method of claim 3, wherein the patient experienced a physical blow to the head within 24 hours prior to the detection. (Item 5) 5. The method according to any one of items 1 to 4, wherein the patient has not been subjected to direct physical or optical impact on the eye. (Item 6) 6. The method of any one of items 1 to 5, wherein the patient is not known or suspected to have Alzheimer's disease. (Item 7) 7. The method according to any one of items 1 to 6, wherein the patient is a human under the age of 40. (Item 8) 8. The method according to any one of items 1 to 7, wherein the detection comprises contacting the amyloid beta protein with a probe in vivo. (Item 9) 9. The method of claim 8, wherein the contact, when activated by light, causes emission of a detectable signal. (Item 10) Item 11. The method according to item 9, wherein the detectable signal is a fluorescent or infrared signal. The probe comprises a compound of formula Ic, [ka] During the ceremony, EDG, a) optionally, one or more R 17or b) -NR 10 R 11 and In the formula, each R 17 are independently halogen, -OR 18 , -NR 19 R 20 , C1~C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons; R 10 , R 11 , R 18 , R 19 and R 20 each independently represents hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less, each of which, other than hydrogen, is optionally represented by one or more R 21 is replaced by R 21 each independently represents a halogen, -OR 22 , -NR 23 R 24 , C1~C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less, wherein said alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene is optionally further represented by one or more R 25 is replaced by R 22 , R 23 and R 24 Each of is independently hydrogen or C1-C 10 is alkyl, R 25 Each of C1 to C10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons; Ar is arylene of up to 14 carbon atoms or heteroarylene of up to 14 carbon atoms, each of which is optionally substituted with one or more R1; Each R1 is independently halogen, -OR2, -NR3R4, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons, wherein said alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene is optionally substituted with one or more R; R2, R3, and R4 are independently hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less, each of which, other than hydrogen, is optionally substituted with one or more R; Each R5 is independently halogen, -OR6, -NR7R8, C1 to C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons; R6, R7, R8 and R 84 are independently hydrogen, C1-C 10 is alkyl, EWG is -F, -Cl, -Br, -CH=O, NO2, -CF3, -CCl3, selected from the group consisting of -SO3H and -CN; WSG, i) [ka] or ii) polyethylene glycol, polypropylene glycol, copolymers of polyethylene glycol and polypropylene glycol, or alkoxy derivatives thereof; iii) [ka] (In the formula, n is an integer of 1 to 50, and R 81 But hydrogen, C1-C 10 Alkyl, C1-C 10 Alkenyl, or C1-C 10 alkynyl, wherein the alkyl, alkenyl, or alkynyl is optionally one or more C1-C 10 Alkyl, C1-C 10 substituted with heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less; iv) [ka] or v) [ka] or vi) -(C1~C 10 Alkyl)-R 33 -R 37 (In the formula, R 33 is a heteroarylene of 10 or less carbons; R 37 is -(C1-C6 alkyl)(heterocycloalkyl of 10 or less carbons) vii) [ka] or viii) -(C1~C 10 Heteroalkyl)-R 33 -R 37 (In the formula, R 33 is a heteroarylene of 10 or less carbons; R 37 is -(C1-C6 alkyl)(heterocycloalkyl of 10 or less carbons); or ix) [ka] and X is C=O or SO2, or X and R 84 and combine to form pyridinyl, 11. The method according to any one of items 8 to 10, wherein Y is NH or S. (Item 12) 11. The method according to any one of items 8 to 10, wherein the probe comprises an antibody. (Item 13) 13. The method of any one of items 1 to 12, further comprising determining that the patient is suffering from TBI if amyloid beta protein is detected in the eye. (Item 14) 14. The method of claim 13, further comprising instructing the patient to refrain from vigorous physical activity. (Item 15) 14. The method of claim 13, further comprising administering to the patient an agent that treats or ameliorates TBI. (Item 16) 1. A method for 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. (Item 17) 17. The method of claim 16, further comprising detecting the binding of the probe to the amyloid beta protein in the eye. (Item 18) 18. The method according to claim 16 or 17, wherein the bond, when activated by light, causes emission of a detectable signal. (Item 19) 19. The method of claim 18, wherein the signal is a fluorescent or infrared signal. (Item 20) The probe is Comprising a compound of formula Ic [ka] During the ceremony, EDG, a) optionally, one or more R 17 or b) -NR 10 R 11 and In the formula, each R 17 are independently halogen, -OR 18 , -NR 19 R 20 , C1~C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons; R 10 , R 11 , R 18 , R 19 and R 20 each independently represents hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less, each of which, other than hydrogen, is optionally represented by one or more R21 is replaced by R 21 each independently represents a halogen, -OR 22 , -NR 23 R 24 , C1~C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less, wherein said alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene is optionally further represented by one or more R 25 is replaced by R 22 , R 23 and R 24 Each of is independently hydrogen or C1-C 10 is alkyl, R 25 Each of C1 to C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons; Ar is arylene of up to 14 carbon atoms or heteroarylene of up to 14 carbon atoms, each of which is optionally substituted with one or more R1; Each R1 is independently halogen, -OR2, -NR3R4, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons, wherein said alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene is optionally substituted with one or more R; R2, R3, and R4 are independently hydrogen, C1-C 10 Alkyl, C1-C 10heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less, each of which, other than hydrogen, is optionally substituted with one or more R; Each R5 is independently halogen, -OR6, -NR7R8, C1 to C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons; R6, R7, R8 and R 84 are independently hydrogen, C1-C 10 is alkyl, EWG is selected from the group consisting of -F, -Cl, -Br, -CH=O, NO2, -CF3, -CCl3, -SO3H and -CN; WSG, i) [ka] or ii) polyethylene glycol, polypropylene glycol, copolymers of polyethylene glycol and polypropylene glycol, or alkoxy derivatives thereof; iii) [ka] (In the formula, n is an integer of 1 to 50, and R 81 But hydrogen, C1-C 10 Alkyl, C1-C 10 Alkenyl, or C1-C 10 alkynyl, wherein the alkyl, alkenyl, or alkynyl is optionally one or more C1-C 10 Alkyl, C1-C 10substituted with heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less; iv) [ka] or v) [ka] or vi) -(C1~C 10 Alkyl)-R 33 -R 37 (In the formula, R 33 is a heteroarylene of 10 or less carbons; R 37 is -(C1-C6 alkyl)(heterocycloalkyl of 10 or less carbons) vii) [ka] or viii) -(C1~C 10 Heteroalkyl)-R 33 -R 37 (In the formula, R 33 is a heteroarylene of 10 or less carbons; R 37 is -(C1-C6 alkyl)(heterocycloalkyl of 10 or less carbons); or ix) [ka] and X is C=O or SO2, or X and R 84 and combine to form pyridinyl, 20. The method according to any one of items 16 to 19, wherein Y is NH or S. (Item 21) 21. The method of any one of items 16 to 20, wherein said administration is intravenous or localized to the retina of said eye. (Item 22) 22. The method according to any one of items 16 to 21, wherein the patient has suffered a physical blow to the head within 30 days prior to the detection. (Item 23) 23. The method of any one of items 16 to 22, wherein the patient is not known or suspected to have Alzheimer's disease. (Item 24) 24. The method of any one of items 16 to 23, wherein the patient has not been subjected to direct physical or optical impact on the eye. (Item 25) A kit or package comprising a probe that specifically binds to amyloid beta protein and a retinal imaging device. (Item 26) Item 26. The kit or package of item 25, wherein the retinal imaging device is equipped with a laser light source. (Item 27) 26. The kit or package of item 25, wherein the retinal imaging device further comprises a retinal scanner. (Item 28) The probe comprises a compound of formula Ic, [ka] During the ceremony, EDG, a) optionally, one or more R 17 or b) -NR 10 R 11 and In the formula, each R 17 are independently halogen, -OR 18 , -NR 19 R 20 , C1~C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons; R 10 , R 11 , R 18 , R 19 and R 20 each independently represents hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less, each of which, other than hydrogen, is optionally represented by one or more R 21 is replaced by R 21 each independently represents a halogen, -OR 22 , -NR 23 R 24 , C1~C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less, wherein said alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene is optionally further represented by one or more R 25 is replaced by R 22 , R 23 and R 24 Each of is independently hydrogen or C1-C 10 is alkyl, R 25 Each of C1 to C 10 Alkyl, C1-C 10heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons; Ar is arylene of up to 14 carbon atoms or heteroarylene of up to 14 carbon atoms, each of which is optionally substituted with one or more R1; Each R1 is independently halogen, -OR2, -NR3R4, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons, wherein said alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene is optionally substituted with one or more R; R2, R3, and R4 are independently hydrogen, C1-C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less, each of which, other than hydrogen, is optionally substituted with one or more R; Each R5 is independently halogen, -OR6, -NR7R8, C1 to C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons; R6, R7, R8 and R 84 are independently hydrogen, C1-C 10 is alkyl, EWG is selected from the group consisting of -F, -Cl, -Br, -CH=O, NO2, -CF3, -CCl3, -SO3H and -CN; WSG, i) [ka] or ii) polyethylene glycol, polypropylene glycol, copolymers of polyethylene glycol and polypropylene glycol, or alkoxy derivatives thereof; iii) [ka] (In the formula, n is an integer of 1 to 50, and R 81 But hydrogen, C1-C 10 Alkyl, C1-C 10 Alkenyl, or C1-C 10 alkynyl, wherein the alkyl, alkenyl, or alkynyl is optionally one or more C1-C 10 Alkyl, C1-C 10 substituted with heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less; iv) [ka] or v) [ka] or vi) -(C1~C 10 Alkyl)-R 33 -R 37 (In the formula, R 33 is a heteroarylene of 10 or less carbons; R 37 is -(C1-C6 alkyl)(heterocycloalkyl of 10 or less carbons) vii) [ka] or viii) -(C1~C 10 Heteroalkyl)-R 33 -R 37 (In the formula, R 33 is a heteroarylene of 10 or less carbons; R 37 is -(C1-C6 alkyl)(heterocycloalkyl of 10 or less carbons); or ix) [ka] and X is C=O or SO2, or X and R 84 and combine to form pyridinyl, 28. The kit or package according to any one of items 25 to 27, wherein Y is NH or S. (Item 29) The probe is [ka] The method according to any one of items 8 to 10, 13 to 19, or 21 to 24, comprising a compound selected from: (Item 30) The probe is [ka] 28. The kit or package according to any one of items 25 to 27, comprising a compound selected from the group consisting of [Brief description of the drawings]

[0023] [Figure 1]Shown are ex vivo fluorescence images of the surface of flat mounted human retina (upper temporal lobe) stained with DAPI (4',6-diamidino-2-phenylindole) and Compound 1. A) Braak stage V / VI AD patient. B) Braak stage V / VI AD patient. C) Cognitively normal patient. D) Cognitively normal patient. Staining is as follows: blue = DAPI nuclear staining, red = autofluorescence, green / yellow = staining with Compound 1. Scale bar = 100 μm. [Diagram 2] Top: Images showing that Compound 1 causes amyloid beta to fluoresce in retinal tissue of a TBI blast mouse model. Bottom: No visible fluorescence in normal mice. Staining is as follows: green = stained with Compound 1, red = stained with 6E10 antibody, an antibody specific for amyloid beta. [Diagram 3] Top: Images showing that Compound 1 fluoresces amyloid beta in brain tissue from a TBI blast mouse model. Bottom: No visible fluorescence in normal mice. Staining is as follows: green = stained with Compound 1, red = stained with 6E10 antibody, an antibody specific for amyloid beta. [Figure 4] In the top row, images are presented showing retinal tissue from a C5BL mouse subjected to controlled cortical impact (CCI) stained with A) DAPI, B) compound 1, C) 6E10 antibody, and D) a merged image of the three stains. Bottom row: E) Retinal tissue from a C5BL / 6 mouse subjected to craniotomy stained with DAPI, compound 1, and 6E10 antibody. Fluorescence spectra of F) DAPI, G) compound 1, H) 6E10 antibody, and I) merged are shown on the left. Arrows indicate visible aggregated proteins. [Diagram 5] In the top row, images are presented showing retinal tissue from a C5BL / 6 mouse undergoing CCI stained with A) DAPI, B) compound 1, C) 6E10 antibody, and D) a merged image of the three stains. In the bottom row: D) merged image of the three stains, E) fluorescence spectrum of DAPI, F) fluorescence spectrum of compound 1, G) fluorescence spectrum of 6E10 antibody, and H) merged fluorescence spectrum of all three stains. [Figure 6]Top: Images showing retinal tissue from a C5BL / 6 mouse subjected to CCI stained with A) DAPI, B) Compound 1, C) 6E10 antibody, D) merged image of the three stains. Bottom: Images showing retinal tissue from a C5BL / 6 mouse subjected to craniotomy stained with E) DAPI, F) Compound 1, G) 6E10 antibody, H) merged image of the three stains. Arrows indicate visible aggregated protein. [Figure 7] 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 from a C5BL / 6 mouse undergoing CCI and stained with DAPI, Compound 1, and 6E10 antibody. [Figure 8] Images of retinal staining from mice subjected to controlled cortical impact (CCI) (top) or sham (bottom) mice are presented. Top: Flat mount images of CCI mouse retinas stained with Compound 1, 6E10, and merged images. Arrows indicate areas of colocalization with Compound 1 and 6E10. Bottom: Images of sham mice stained with Compound 1, 6E10, and merged images. [Figure 9] Images of live retinal imaging with Compound 23 in mice before and after CCI are presented. Top: Time-lapse retinal imaging of a 3-month-old mouse pre-CCI after iv administration of Compound 23. Labeled time courses at the top indicate the time points of in vivo retinal imaging before (t=0) and after iv administration of Compound 23. Bottom: Time-lapse retinal imaging of the same mouse 24 hours after CCI with Compound 23.

[0024] It will be appreciated that some or all of the figures are schematic for illustrative purposes. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0025] definition The following description describes exemplary embodiments of the present technology, however, it should be recognized that such description is not intended as a limitation on the scope of the present disclosure, but is instead provided as a description of exemplary embodiments.

[0026] As used herein, the following words, phrases, and symbols are intended to have the meanings generally set forth below, except to the extent that the context in which they are used indicates otherwise.

[0027] The term “alkyl,” by itself or as part of another substituent, refers to straight-chain (i.e., unbranched) or branched chain, or combinations thereof, which may be fully saturated, mono- or polyunsaturated, and has a specified number of carbon atoms (i.e., C1 to C6). 10 means 1 to 10 carbons. Examples of saturated hydrocarbon radicals include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, (cyclohexyl)methyl, homologs and isomers of, e.g., n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like.

[0028] The term "heteroalkyl," by itself or in combination with another term, means, unless otherwise stated, a stable linear or branched chain, or cyclic hydrocarbon radical, or combination thereof, consisting of at least one carbon atom and at least one heteroatom selected from the group consisting of O, N, P, Si, and S, the nitrogen and sulfur atoms of which 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 group or at the position at which the alkyl group is attached to the remainder 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-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 be consecutive.

[0029] The terms "cycloalkyl" and "heterocycloalkyl", by themselves or in combination with other terms, represent cyclic versions of "alkyl" and "heteroalkyl", respectively, unless otherwise specified. Additionally, in the case of heterocycloalkyl, a heteroatom can occupy the position at which the heterocycle is attached to the remainder of the molecule. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples of heterocycloalkyl include, but are not limited to, tetrahydropyran, 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl, 2-piperazinyl, and the like. Examples of heterocycloalkyls include, but are not limited to, glucose, mannose, allose, altrose, gulose, idose, galactose, and talose. [ka] These include, but are not limited to:

[0030] The terms "halo" or "halogen," by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom. Additionally, terms such as "haloalkyl" are meant to include monohaloalkyl and polyhaloalkyl. For example, the term "halo(C1-C4)alkyl" is meant to include, but is not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0031] The term "aryl," unless otherwise specified, means a polyvalent unsaturated aromatic hydrocarbon substituent that may be monocyclic or polycyclic (preferably 1-3 rings) fused together (i.e., fused ring aryl) or covalently linked together. Fused ring aryl refers to multiple rings fused together, at least one of the fused rings being an aryl ring.

[0032] The term "heteroaryl" refers to an aryl group (or ring) containing 1 to 4 heteroatoms selected from N, O, and S, where the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atom is optionally quaternized. Thus, the term "heteroaryl" includes fused ring heteroaryl groups (i.e., multiple rings fused together, where at least one of the fused rings is a heteroaromatic ring). 5,6-fused ring heteroarylene refers to two rings fused together, where one ring has five ring members and the other ring has six ring members, and where at least one ring is a heteroaryl ring. Similarly, 6,6-fused ring heteroarylene refers to two rings fused together, where one ring has six ring members and the other ring has six ring members, and where at least one ring is a heteroaryl ring. and 6,5-fused ring heteroarylene refers to two rings fused together, one ring having six ring members and the other ring having five ring members, with at least one ring being a heteroaryl ring. The heteroaryl group can be bonded to the remainder of the molecule through a 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 ...

[0033] Examples of aryl and heteroaryl ring systems include aryl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 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 aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below.

[0033] An "arylene" and a "heteroarylene," alone or as part of another substituent, mean a divalent radical derived from an aryl and heteroaryl, respectively.

[0034] For brevity, the term "aryl" when used in combination with other terms (e.g., aryloxy, arylthioxy, arylalkyl) includes both aryl and heteroaryl rings as defined above. Thus, the term "arylalkyl" is meant to include radicals in which an aryl group is attached to an alkyl group (e.g., benzyl, phenethyl, pyridylmethyl, etc.), including alkyl groups in which a carbon atom (e.g., a methylene group) has been replaced by, for example, an oxygen atom (e.g., phenoxymethyl, 2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, etc.).

[0035] Each of the above terms (e.g., "alkyl," "heteroalkyl," "aryl" and "heteroaryl") are meant to include both substituted and unsubstituted forms of the indicated radical. Preferred substituents for each type of radical are provided below.

[0036] As used herein, the term "heteroatom" or "ring heteroatom" is meant to include oxygen (O), nitrogen (N), sulfur (S), phosphorus (P), and silicon (Si).

[0037] Certain commonly used alternative chemical names may be used. For example, divalent groups such as divalent "alkyl" groups, divalent "aryl" groups, etc. may be referred to as "alkylene" or "alkylenyl" groups, "arylene" or "arylenyl" groups, respectively. Also, unless otherwise indicated, when a combination of groups is referred to herein as a single moiety, e.g., arylalkyl, the last-mentioned group contains the atom by which that moiety is attached to the remainder of the molecule.

[0038] The term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes both the occurrence and non-occurrence of the event or circumstance. Also, the term "optionally substituted" refers to any one or more hydrogen atoms on a specified atom or group that may or may not be replaced by a non-hydrogen moiety.

[0039] Some compounds exist as tautomers. Tautomers are in equilibrium with each other. For example, amide-containing compounds may exist in equilibrium with imidic acid tautomers. Regardless of which tautomer is shown and regardless of the nature of the equilibrium between tautomers, the compounds are understood by those skilled in the art to include both amide and imidic acid tautomers. Thus, amide-containing compounds are understood to include their imidic acid tautomers. Similarly, imidic acid-containing compounds are understood to include their amide tautomers.

[0040] Any formula or structure given herein is also intended to represent unlabeled form of the compound as well as isotopically labeled form.Isotopically labeled compounds have the structure shown by the formula given herein, except that one or more atoms are replaced by an atom with a selected atomic weight or mass number.Examples of isotopes that can be incorporated into the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, but 2 H (deuterium, D), 3 H (tritium), 11 C. 13 C. 14 C. 15 N, 18 F, 31 P, 32 P, 35 S, 36 Cl and 125 I. Various isotopically labeled compounds of the present disclosure, such as 3 H, 13 C, and 14Those incorporating a radioactive isotope such as C. Such isotopically labeled compounds may be 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 in radiotherapy of patients.

[0041] The present disclosure also includes "deuterated analogs" of compounds of formula I in which one to n hydrogens attached to a carbon atom are replaced with deuterium, where n is the number of hydrogens in the molecule. Such compounds exhibit increased resistance to metabolism and are therefore useful for increasing the half-life of any compound of formula I when administered to a mammal, particularly a human. 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 means well known in the art, for example, by using starting materials in which one or more hydrogens have been replaced by deuterium.

[0042] Deuterium-labeled or substituted therapeutic compounds of the present disclosure may have improved DMPK (drug metabolism and pharmacokinetic) properties related to distribution, metabolism, and excretion (ADME). Substitution with heavier isotopes such as deuterium may provide certain therapeutic advantages resulting from higher metabolic stability, such as increased in vivo half-life, reduced dosage requirements, and / or improved therapeutic index. 18 F-labeled compounds can be useful for PET or SPECT studies.The isotopically labeled compounds and their prodrugs of the present disclosure can generally be prepared by carrying out the procedures disclosed in the schemes or in the examples and preparations described below, by replacing readily available isotopically labeled reagents with non-isotopically labeled reagents.It is understood that deuterium in this context is considered as a substituent in the compounds described herein.

[0043] The concentration of such heavier isotopes, specifically deuterium, can be defined by the isotopic enrichment factor. In the compounds of the present disclosure, any atom not specifically designated as a particular isotope is meant to represent any stable isotope of that atom. Unless otherwise stated, when a position is specifically designated as "H" or "hydrogen", that position is understood to have hydrogen in its natural abundance isotopic composition. Thus, in the compounds of the present disclosure, any atom specifically designated as deuterium (D) is meant to represent deuterium.

[0044] 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.

[0045] Pharmaceutically acceptable salts, hydrates, solvates, tautomeric forms, polymorphs, and prodrugs of the compounds described herein are also provided. "Pharmaceutically acceptable" or "physiologically acceptable" refers to compounds, salts, compositions, dosage forms, and other materials useful in preparing pharmaceutical compositions suitable for veterinary or human medical use.

[0046] 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 salt" or "physiologically acceptable salt" includes, for example, salts with inorganic acids and salts with organic acids. In addition, when a compound described herein is obtained as an acid addition salt, the free base can be obtained by basifying a solution of the acid salt. Conversely, when the product is a free base, an addition salt, particularly a pharmaceutically acceptable addition salt, can be prepared by dissolving the free base in a suitable organic solvent and treating the solution with an acid according to conventional procedures for preparing acid addition salts from basic compounds. Those skilled in the art will recognize various synthetic methodologies that can be used to prepare non-toxic pharmaceutically acceptable addition salts. Pharmaceutically acceptable acid addition salts can be prepared from inorganic and organic acids. Salts 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-toluene-sulfonic acid, salicylic acid, and the like. Similarly, pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases. Salts derived from inorganic bases include, by way of example only, sodium, potassium, lithium, ammonium, calcium, and magnesium salts.Salts derived from organic bases include, for example, alkyl amines (i.e., NH2(alkyl)), dialkyl amines (i.e., HN(alkyl)2), trialkyl amines (i.e., N(alkyl)3), substituted alkyl amines (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), alkenyl amines (i.e., NH2(alkenyl)), dialkenyl amines (i.e., HN(alkenyl)2), trialkenyl amines (i.e., N(alkenyl)3), substituted alkenyl amines (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), Suitable amines include, but are not limited to, primary, secondary, and tertiary amines such as mono-, di-, or tri-cycloalkylamines (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. 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.

[0047] The term "substituted" means that any one or more hydrogen atoms on the designated atom or group are replaced with one or more substituents other than hydrogen, provided that the normal valence of the designated atom is not exceeded. The one or more substituents include, but are not limited to, alkyl, alkenyl, alkynyl, alkoxy, acyl, amino, amido, amidino, aryl, azido, carbamoyl, carboxyl, carboxyl ester, cyano, guanidino, halo, haloalkyl, haloalkoxy, heteroalkyl, heteroaryl, heterocyclyl, hydroxy, hydrazino, imino, oxo, nitro, alkylsulfinyl, sulfonic acid, alkylsulfonyl, thiocyanate, thiol, thione, or combinations thereof. Polymers or similar indefinite structures arrived at by defining a substituent with an infinite number of additional substituents added (e.g., substituted aryl with a substituted alkyl, which is itself substituted with a substituted aryl group, which is further substituted with a substituted heteroalkyl group, etc.) are not intended to be included herein. Unless otherwise specified, the maximum number of consecutive substitutions in the compounds described herein is three. For example, the sequential substitution of a substituted aryl group with two other substituted aryl groups is limited to ((substituted aryl)substituted aryl)substituted aryl. Similarly, the above definition is not intended to include impermissible substitution patterns (e.g., methyl substituted with five fluorines or a heteroaryl group with two adjacent oxygen ring atoms). Such impermissible substitution patterns are well known to those of skill in the art. When used to modify a chemical group, the term "substituted" may describe other chemical groups as defined herein. Unless otherwise specified, when a group is described as being optionally substituted, the substituents of the group are themselves unsubstituted. For example, in some embodiments, the term "substituted alkyl" refers to an alkyl group having one or more substituents, including hydroxyl, halo, alkoxy, cycloalkyl, heterocyclyl, aryl, and heteroaryl. In other embodiments, one or more of the substituents may be further substituted with halo, alkyl, haloalkyl, hydroxyl, alkoxy, cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is substituted.In other embodiments, the substituents may be further substituted with halo, alkyl, haloalkyl, alkoxy, hydroxyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is unsubstituted.

[0048] 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 retarding agents, etc. The use of such media and agents for pharmaceutically active substances is well known in the art. Except where a conventional media or agent is incompatible with the active ingredient, its use in the therapeutic composition is contemplated. Auxiliary active ingredients can also be incorporated into the composition.

[0049] A "solvate" is formed by the interaction of a solvent with a compound. Solvates of the salts of the compounds described herein are also provided. Hydrates of the compounds described herein are also provided.

[0050] Detecting Traumatic Brain Injury (TBI) It is contemplated that traumatic brain injury (TBI), even mild TBI, can rapidly cause the release and / or accumulation of amyloid beta (Aβ) in the retina of a patient. As the experimental examples show, the presence of such Aβ in the retina can be detected with a probe capable of binding to Aβ, and the binding can then be detected by means such as laser-activated fluorescent scanning of the retina.

[0051] A "traumatic brain injury" (TBI) occurs when a crash, blow, impact, or other head injury causes injury to the brain. TBI can lead to permanent brain damage or death. Half of all TBIs are the result of car accidents. Military personnel in combat zones are also at risk. Symptoms of 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 fatigue. People with moderate or severe TBI may experience symptoms such as a worsening or non-resolving headache, repeated vomiting or nausea, convulsions or seizures, difficulty waking from sleep, slurred speech, weakness or numbness in the arms and legs, and dilated pupils.

[0052] "Amyloid beta" (Aβ or Abeta) refers to a peptide of about 36-43 amino acids that is involved in Alzheimer's disease (AD) as the main component of amyloid plaques found in the brains of Alzheimer's patients. The peptide is derived from the amyloid precursor protein (APP, e.g., GenBank accession number: NP_000475), which is cleaved by beta-secretase and gamma-secretase to yield Aβ. Aβ molecules can aggregate to form flexible soluble oligomers that can exist in several forms. It is believed that certain misfolded oligomers (known as "seeds") can induce other Aβ molecules to also adopt the misfolded oligomeric form, setting off a chain reaction similar to prion infection. The oligomers are toxic to neuronal cells.

[0053] Therefore, according to one embodiment of the present disclosure, a method is provided for determining whether a patient suffers from traumatic brain injury (TBI).The method involves detecting the presence of amyloid beta protein in the eye of the patient.It is contemplated that the accumulation of amyloid beta protein occurs mainly in the retina.Therefore, detection can be mainly targeted to the amyloid beta protein in the retina of the eye.

[0054] It is contemplated that amyloid beta protein accumulation may begin soon after a physical impact to the head that causes TBI. It is understood that physical impact may not be the only cause of TBI. Sound, light, and temperature may also cause TBI and are therefore within the scope of this disclosure. In some embodiments, detection is performed within 1 hour or 2 hours of the potential cause of TBI. Alternatively, detection can be performed within 6, 12, 18, 24, 36, or 48 hours. In some embodiments, detection is performed at least 1, 2, 3, 4, 6, 8, 12, 18, or 24 hours after the potential cause of TBI. In some embodiments, detection is performed at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 days after the potential cause of TBI. In some embodiments, detection is performed within 3, 4, 5, 6, 7, 8, 9, or 10 days, or within 1, 2, 3, 4, or 5 weeks, after the potential cause of TBI.

[0055] In some embodiments, the patient may suffer from another disease or condition that causes the accumulation of amyloid beta protein.An example of such a disease is Alzheimer's disease (AD).However, it is contemplated that the patient of the presently disclosed detection method does not suffer from such other disease or condition (e.g., AD).In some embodiments, the patient is young enough (e.g., under 70, 65, 60, 55, 50, 45 or 40 years old) that it is not reasonably suspected that the patient suffers from such disease or condition.

[0056] Direct physical impact (or light, sound, or temperature) on the eye can cause the release or accumulation of amyloid beta protein in the eye. In some embodiments, the patient of this method is not affected by such direct impact on the eye.

[0057] In another embodiment, a method for preparing a patient for diagnosis of traumatic brain injury (TBI) is provided, comprising administering a probe that specifically binds to amyloid beta protein to the patient's eye.The types of patients suitable for such methods are described above, but are not limited thereto.When the probe is administered to the patient's eye, its binding to amyloid beta protein can be detected by the method described herein, and the binding indicates the accumulation of amyloid beta protein, which is an indicator of traumatic brain injury.

[0058] Amyloid beta binding probes and detection The detection of amyloid beta protein can be carried out using a probe that can selectively bind to amyloid beta protein, which is referred to herein as amyloid beta binding (Aβ binding) probe. Such probes are known or can be easily developed or prepared.

[0059] In one embodiment, the Aβ-binding probe, upon binding to Aβ, can be detected via its emitted fluorescent signal upon activation by laser light. Examples of Aβ-binding fluorescent probes are provided in the following section.

[0060] 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 a probe that can bind to the antibody and emit a detectable signal. Non-limiting examples of antibodies include the 6E10 antibody used herein and ab2539 available from Abcam (Cambridge, MA).

[0061] The in situ detection of the binding of Aβ-binding probe to Aβ in the retina of a patient can be facilitated by using a retinal imaging device, which is preferably handheld or portable. The retinal imaging device can 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 in the retina and binds to the Aβ-binding probe, the accumulation can be easily detected and quantified by the lens and image sensor that collects and senses the fluorescent signal.

[0062] Amyloid-sensitive fluorescent probe (ASF) The present disclosure further provides a small molecule fluorescent probe that can bind to amyloid beta (Aβ). In one embodiment, the probe can be selected from compounds as described in WO2011 / 072257, WO2015 / 143185, or WO2017 / 004560, which are incorporated by reference in their entirety. In one embodiment, the probe can be curcumin or other compounds currently used to stain Aβ, such as thioflavin or Congo Red.

[0063] In one embodiment, the probe may be selected from the compounds described in WO2015 / 143185, which compounds are described below.

[0064] In certain embodiments, the present disclosure provides a compound of formula I, or a salt or solvate thereof: [ka] During the ceremony, EDG is an electron donating group, Each Ar is independently selected from C1 to C6, each of which is optionally substituted with one or more R1. 14 Arylene or C1-C 14 is heteroarylene, Each R1 is independently halogen, -OR2, -NR3R4, C1-C 10 Alkyl, C1-C 10Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene, or C1-C 10 is heteroarylene, 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, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene, or C1-C 10 heteroarylene, each of which, except for hydrogen, is optionally substituted with one or more R5; Each R5 is independently selected from halogen, -OR6, -NR7R8, C1-C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene, or C1-C 10 is heteroarylene, R6, R7, R8 and R 84 are independently hydrogen, C1 to C 10 is alkyl, R 84 is hydrogen or C1-C 10 is alkyl, EWG is an electron withdrawing group, WSG is a water-soluble group, X is C=O or SO2, or X and R 84 and combine to form pyridinyl, Y is NH or S; each x is independently an integer from 0 to 10; each w is independently an integer from 1 to 5; each y is independently an integer from 0 to 10; z is an integer from 1 to 10.

[0065] In certain embodiments, the compound is of formula II, or a salt or solvate thereof: [ka] , During the ceremony, EDG is an electron donating group, Ar2 and each Ar1 are independently C1 to C 14 Arylene or C1-C 14 is heteroarylene, Each optionally contains one or more R 41 is replaced by 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 is heteroarylene, The alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene may optionally be one or more R 45 is replaced by 、 R 42 , R 43 and R 44 are independently hydrogen, C1 to 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 for hydrogen, is optionally represented by one or more R 45 is replaced by Each R 45are independently halogen, -OR 46 , -NR 47 R 48 , 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 46 , R 47 and R 48 are independently hydrogen or C1-C 10 is alkyl, EWG is an electron withdrawing group, Y is absent, O, NH, or S; WSG is hydrogen or a water soluble group; x is an integer from 0 to 10; y is an integer from 0 to 10; and z is an integer from 1 to 10.

[0066] In certain embodiments, R 84 is hydrogen. In certain embodiments, R 84 is C1~C 10 In certain embodiments, R 84 is methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, neptyl, or decyl. 84 is methyl.

[0067] The substituent EDG, as known in the art, is an electron donating group. In certain embodiments, EDG is any atom or functional group that can donate a portion of its electron density to a conjugated pi system, thus making the pi system more nucleophilic.

[0068] In certain embodiments, EDG, -OR9, -NR 10 R 11 , -SR 12 , -PR 13R 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 is heteroarylene, The alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene may optionally be one or more R 17 is replaced by 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 is heteroarylene, R9, R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 18 , R 19 , R 20 Each of the groups is independently hydrogen, C1 to C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene, or C1-C 10 is heteroarylene, Each of these, except hydrogen, may optionally be one or more R 21 is replaced by R 10 and R 11 are optionally joined together and optionally R 21forming a heterocycloalkyl or heteroaryl substituted with R 21 Each of the groups is independently a halogen, -OR 22 , N.R. 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, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene is optionally further represented by one or more R 25 is replaced by R 22 , R 23 and R 24 Each of is independently hydrogen or C1-C 10 is alkyl, Each R 25 are independent, C1~C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene, or C1-C 10 It is a heteroarylene.

[0069] In certain embodiments, the EDG is selected from the group consisting of: [ka]

[0070] In certain embodiments, the EDG is [ka] In certain embodiments, the EDG is [ka] It is.

[0071] EWG is an electron withdrawing group. In certain embodiments, an electron withdrawing group as used herein is any atom or group that can attract electron density from adjacent atoms towards itself, either by resonance or inductive effects.

[0072] In certain embodiments, EWG is halogen, -CN, -NO2, -SO3H, -CR 26 R 27 R 28 , -COR 29 , or -COOR 30 is selected from the group consisting of Each R 26 , R 27 and R 28 are independently hydrogen or halogen; R 29 are halogens, 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 is heteroarylene, An alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene may optionally be represented by one or more R 31 is replaced by 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 is heteroarylene, The alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene may optionally be one or more R 32is replaced by Each R 31 and R 32 are C1 to C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene, or C1-C 10 It is a heteroarylene.

[0073] In certain embodiments, the EWG is -F, -Cl, -Br, Selected from the group consisting of -CH=O, NO2, -CF3, -CCl3, -SO3 and -CN. In certain embodiments, the EWG is F, Cl, or Br. In certain embodiments, the EWG is -CN.

[0074] WSG is a water-soluble group. In certain embodiments, the WSG group serves to modify the solubility of the compound in aqueous systems.

[0075] In certain embodiments, 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 is heteroarylene, The alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene may optionally be one or more R 33 is replaced by 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-C10 Arylene, or C1-C 10 heteroarylene, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene is optionally further represented by one or more R 37 is replaced by Each R 34 , R 35 and R 36 are independently hydrogen, C1 to C 10 Alkyl, C1-C 10 Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene, or C1-C 10 is heteroarylene, The alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene may optionally be one or more R 37 is replaced by 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, -(C1-C6 alkyl)(C1-C 10 Heterocycloalkyl), C1-C 10 Arylene, or C1-C 10 is heteroarylene, R 38 , R 39 and R 40 Each of is independently hydrogen or C1-C 10 It is an alkyl.

[0076] In one particular embodiment, the WSG [ka] It is.

[0077] In certain embodiments, the WSG is polyethylene glycol, polypropylene glycol, a copolymer of polyethylene glycol and polypropylene glycol, or an alkoxy derivative thereof.

[0078] In one particular embodiment, the WSG [ka] In the formula, 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, and in each occurrence, alkyl, alkenyl, or alkynyl optionally includes 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 is hydrogen. In certain embodiments, R 81 is methyl. In certain embodiments, R 81 is ethyl. In certain embodiments, R 81 teeth [ka] 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 value of 1 to 10, 1 to 20, 1 to 30, 1 to 40, 1 to 50, 10 to 20, 10 to 30, 10 to 40, 10 to 50, 20 to 30, 20 to 40, 20 to 50, 30 to 40, 30 to 50, or 40 to 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.

[0079] In certain embodiments, R 81 is hydrogen.

[0080] In one particular embodiment, the WSG [ka] It is.

[0081] In one particular embodiment, the WSG [ka] It is.

[0082] In one particular embodiment, the WSG [ka] It is.

[0083] In one particular embodiment, the WSG [ka] It is.

[0084] In one particular embodiment, the WSG [ka] where each R 82 are independently hydrogen or C1-C 10 It is an alkyl.

[0085] In certain embodiments, each R 82 is independently hydrogen, methyl, ethyl, propyl, or butyl.

[0086] In one particular embodiment, the WSG [ka] It is.

[0087] In one particular embodiment, the WSG [ka] It is.

[0088] In one particular embodiment, the WSG [ka] where each R 83 is hydrogen or C1-C 10 In certain embodiments, each R 83 is independently hydrogen, methyl, ethyl, propyl, or butyl.

[0089] In one particular embodiment, the WSG [ka] It is.

[0090] In one particular embodiment, the WSG [ka] It is.

[0091] In certain embodiments, WSG is -(C 10 Alkylene)-R 33 -R 37 In certain embodiments, WSG is -(C 10 Alkylene)-R 33 -R 37 and R 33 is C1~C 10 In certain embodiments, WSG is -(C 10 Alkyl)-R 33 -R 37 and R 33 is C1~C 10 Heteroarylene, R 37 is -(C1-C6 alkyl)(C1-C 10 In certain embodiments, WSG is -CH-R 33 -R 37 In certain embodiments, WSG is -CH-R 33 -R 37 and R 33 is triazole, imidazole, or pyrazole. In certain embodiments, WSG is -CH-R 33 -R 37 and R 33 is triazole. In certain embodiments, WSG is -CH-R 33 -R 37 and R 33 is 1,2,4-triazole. In certain embodiments, WSG is -CH-R 33 -R 37 and R 33 is 1,2,3-triazole. In certain embodiments, WSG is -CH-R 33 -R 37 and R 33 is 1,2,3-triazole, R37 is -(C1-C6 alkyl)(C1-C 10 In certain embodiments, WSG is -CH-R 33 -R 37 and R 33 is 1,2,3-triazole, R 37 is -(C1 alkyl)(C1-C 10 In certain embodiments, WSG is -CH-R 33 -R 37 and R 33 is 1,2,3-triazole, R 37 is -(C1 alkyl)(C1-C 10 Heterocycloalkyl) C1-C 10 The heterocycloalkyl is a tetrahydropyran derivative.

[0092] In one particular embodiment, the WSG [ka] where each R 87 is hydrogen, C1-C 10 Alkyl, or -C(=O)C1~C 10 In certain embodiments, each R 87 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.

[0093] In one particular embodiment, the WSG [ka] It is.

[0094] In one particular embodiment, the WSG [ka] It is.

[0095] In certain embodiments, WSG is -(C 10 Heteroalkyl)-R 33 -R 37 In certain embodiments, WSG is -(C 10 Heteroalkyl)-R 33 -R 37 and R 33 is C1~C 10 In certain embodiments, WSG is -(C 10 Heteroalkyl)-R 33 -R 37 and R 33 is C1~C 10 Heteroarylene, R 37 is -(C1-C6 alkyl)(C1-C 10 heterocycloalkyl).

[0096] In one particular embodiment, the WSG [ka] 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 value of 1 to 10, 1 to 20, 1 to 30, 1 to 40, 1 to 50, 10 to 20, 10 to 30, 10 to 40, 10 to 50, 20 to 30, 20 to 40, 20 to 50, 30 to 40, 30 to 50, or 40 to 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.

[0097] In one particular embodiment, the WSG [ka] and R 33 is C1~C 10 In certain embodiments, R 33 is C5 heteroarylene. In certain embodiments, R 33 is triazole, imidazole, or pyrazole. In certain embodiments, R 33 is triazole. In 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, R 37 is -(C1-C6 alkyl)(C1-C 10 In certain embodiments, R 33 is 1,2,3-triazole, 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, R 37 teeth [ka] It is.

[0098] In one particular embodiment, the WSG [ka] and R 33 is 1,2,3-triazole, R 37 teeth [ka] and p is 3.

[0099] In one particular embodiment, the WSG [ka] where each R 87 is hydrogen, C1-C 10 Alkyl, or -C(=O)C1~C 10 In certain embodiments, each R 87 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.

[0100] In one particular embodiment, the WSG comprises: [ka] It is.

[0101] In one particular embodiment, the WSG [ka] It is.

[0102] In certain embodiments, X is C=O or SO. In certain embodiments, X is C=O. In certain embodiments, X is SO.

[0103] In certain embodiments, Y is NH or S. In certain embodiments, Y is NH. In certain embodiments, Y is S.

[0104] 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.

[0105] 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.

[0106] 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.

[0107] 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.

[0108] In certain embodiments, x is 0, w is 1, y is 0, z is 1, X is C=O, and Y is NH.

[0109] In certain embodiments, x is 0, w is 1, y is 0, z is 1, X is SO2, and Y is NH.

[0110] In certain embodiments, x is 0, w is 2, y is 0, z is 1, X is C=O, and Y is NH.

[0111] In certain embodiments, x is 0, w is 2, y is 0, z is 1, X is SO2, and Y is NH.

[0112] In certain embodiments, the present disclosure provides a compound of formula Ia: [ka] , where EDG, Ar, R 84 , x, w, y, z, EWG, and WSG are defined as above.

[0113] In certain embodiments, the present disclosure provides a compound of formula Ib: [ka] , where EDG, Ar, R 84, x, w, y, z, EWG, and WSG are defined above.

[0114] In one aspect, the disclosure provides a compound of formula Ic: [ka] , where EDG, Ar, R 84 , X, Y, EWG, and WSG are defined above.

[0115] In certain embodiments, a compound of formula (Ic), or a salt or solvate thereof, [ka] During the ceremony, EDG, a) optionally, one or more R 17 or b) -NR 10 R 11 and In the formula, each R 17 are independently halogen, -OR 18 , -NR 19 R 20 , C1~C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons; R 10 , R 11 , R 18 , R 19 and R 20 Each of the groups is independently hydrogen, C1 to C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less, each of which, other than hydrogen, is optionally represented by one or more R 21is replaced by R 21 Each of the groups is independently a halogen, -OR 22 , -NR 23 R 24 , C1~C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene is optionally further represented by one or more R 25 is replaced by R 22 , R 23 and R 24 Each of is independently hydrogen or C1-C 10 is alkyl, R 25 Each of C1 to C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons; Ar is an arylene of up to 14 carbon atoms or a heteroarylene of up to 14 carbon atoms, each of which is optionally substituted with one or more R1; Each R1 is independently halogen, -OR2, -NR3R4, C1 to C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons, wherein the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene is optionally substituted with one or more R; R2, R3, and R4 are independently hydrogen, C1-C 10 Alkyl, C1-C 10heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less, each of which, other than hydrogen, is optionally substituted with one or more R; Each R5 is independently halogen, -OR6, -NR7R8, C1 to C 10 Alkyl, C1-C 10 heteroalkyl, cycloalkyl of 10 or fewer carbons, heterocycloalkyl of 10 or fewer carbons, arylene of 10 or fewer carbons, or heteroarylene of 10 or fewer carbons; R6, R7, R8 and R 84 are independently hydrogen, C1-C 10 is alkyl, EWG is selected from the group consisting of -F, -Cl, -Br, -CH=O, NO2, -CF3, -CCl3, -SO3H and -CN; WSG is i) [ka] or ii) polyethylene glycol, polypropylene glycol, copolymers of polyethylene glycol and polypropylene glycol, or alkoxy derivatives thereof; iii) [ka] (In the formula, n is an integer of 1 to 50, and R 81 is hydrogen, C1-C 10 Alkyl, C1-C 10 Alkenyl, or C1-C 10 Alkynyl, where the alkyl, alkenyl, or alkynyl is optionally one or more C1-C 10 Alkyl, C1-C 10substituted with heteroalkyl, cycloalkyl of 10 carbons or less, heterocycloalkyl of 10 carbons or less, arylene of 10 carbons or less, or heteroarylene of 10 carbons or less; iv) [ka] or v) [ka] or vi) -(C1~C 10 Alkyl)-R 33 -R 37 (In the formula, R 33 is a heteroarylene of 10 carbons or less, R 37 is -(C1-C6 alkyl)(heterocycloalkyl of 10 or less carbons) vii) [ka] or viii) -(C1~C 10 Heteroalkyl)-R 33 -R 37 (In the formula, R 33 is a heteroarylene of 10 carbons or less, R 37 is -(C1-C6 alkyl)(heterocycloalkyl of 10 or less carbons); or ix) [ka] and X is C=O or SO2, or X and R 84 and combine to form pyridinyl, Y is NH or S. In certain embodiments, R 81 is hydrogen. In one particular embodiment, the WSG comprises: [ka] It is. In one particular embodiment, the WSG comprises: [ka] It is. In one aspect, the disclosure provides a compound of formula Id: [ka] , where EDG, R 84 , Ar, EWG, and WSG are defined as above.

[0116] In one aspect, the disclosure provides a compound of formula Ie: [ka] , where EDG, R 84 , Ar, EWG, and WSG are defined as above.

[0117] In some cases, the compound is [ka] where n is an integer with a value from 1 to 50. In some cases, n is an integer with a value from 1 to 10, e.g., n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0118] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0119] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0120] In certain embodiments, the compound is [ka] where p is an integer having a value from 1 to 50. In some cases, p is an integer having a value from 1 to 10, e.g., p is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0121] In certain embodiments, the compound is [ka] where n is an integer with a value from 1 to 50. In some cases, n is an integer with a value from 1 to 10, e.g., n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0122] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0123] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0124] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0125] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0126] In certain embodiments, the compound is [ka] where n is an integer with a value from 1 to 50. In some cases, n is an integer with a value from 1 to 10, e.g., n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0127] In certain embodiments, the compound is [ka] where n is an integer with a value from 1 to 50. In some cases, n is an integer with a value from 1 to 10, e.g., n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0128] In certain embodiments, the compound is [ka] where n is an integer with a value from 1 to 50. In some cases, n is an integer with a value from 1 to 10, e.g., n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0129] In certain embodiments, the compound is [ka] where n is an integer with a value from 1 to 50. In some cases, n is an integer with a value from 1 to 10, e.g., n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0130] In certain embodiments, the compound is [ka] where n is an integer with a value from 1 to 50. In some cases, n is an integer with a value from 1 to 10, e.g., n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0131] In certain embodiments, the compound is [ka] where n is an integer with a value from 1 to 50. In some cases, n is an integer with a value from 1 to 10, e.g., n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0132] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0133] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0134] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0135] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0136] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0137] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0138] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0139] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0140] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0141] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0142] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0143] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0144] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0145] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0146] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0147] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0148] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0149] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0150] In certain embodiments, the compound is [ka] It is.

[0151] 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.

[0152] In certain embodiments, the compound is [ka] It is.

[0153] In certain embodiments, the compound is (E)-1-cyano-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)ethenesulfonamide. In certain embodiments, the compound is (Z)-1-cyano-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)ethenesulfonamide.

[0154] In certain embodiments, the compound is [ka] It is.

[0155] In certain embodiments, the compound is (E)-2-cyano-N-(2,5,8,11,14,17-hexaoxanonadecan-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-hexaoxanonadecan-19-yl)-3-(6-(piperidin-1-yl)naphthalen-2-yl)acrylamide.

[0156] In certain embodiments, the compound is [ka] It is.

[0157] In certain embodiments, the compound is (E)-1-cyano-N-(2,5,8,11,14,17-hexaoxanonadecane-19-yl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)ethenesulfonamide. In certain embodiments, the compound is (Z)-1-cyano-N-(2,5,8,11,14,17-hexaoxanonadecane-19-yl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)ethenesulfonamide.

[0158] In certain embodiments, the compound is [ka] It is.

[0159] 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.

[0160] In certain embodiments, the compound is [ka] It is.

[0161] In certain embodiments, the compound is (E)-1-cyano-N-(2,3-dihydroxypropyl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)ethenesulfonamide. In certain embodiments, the compound is (Z)-1-cyano-N-(2,3-dihydroxypropyl)-2-(6-(piperidin-1-yl)naphthalen-2-yl)ethenesulfonamide.

[0162] In certain embodiments, the compound is [ka] It is.

[0163] 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.

[0164] In certain embodiments, the compound is [ka] It is.

[0165] 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.

[0166] In certain embodiments, the compound is [ka] It is.

[0167] In certain embodiments, the compound is (E)-2-cyano-N-(2,5,8,11,14,17-hexaoxanonadecane-19-yl)-3-(6-(piperidin-1-yl)naphthalene-2-yl)buta-2-enamide. In certain embodiments, the compound is (Z)-2-cyano-N-(2,5,8,11,14,17-hexaoxanonadecane-19-yl)-3-(6-(piperidin-1-yl)naphthalene-2-yl)buta-2-enamide.

[0168] In certain embodiments, the compound is [ka] It is.

[0169] In certain embodiments, the compound is (E)-1-cyano-N-(2,5,8,11,14,17-hexaoxanonadecane-19-yl)-2-(6-(piperidin-1-yl)naphthalene-2-yl)prop-1-ene-1-sulfonamide. In certain embodiments, the compound is (Z)-1-cyano-N-(2,5,8,11,14,17-hexaoxanonadecane-19-yl)-2-(6-(piperidin-1-yl)naphthalene-2-yl)prop-1-ene-1-sulfonamide.

[0170] In certain embodiments, the compound is [ka] It is.

[0171] 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.

[0172] In certain embodiments, the compound is [ka] It is.

[0173] 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.

[0174] In certain embodiments, the compound is [ka] It is.

[0175] In certain embodiments, the compound is (R,E)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl)-N-((3,4,5,6-tetrahydroxytetrahydro-2H-pyran-2-yl)methyl)acrylamide. 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.

[0176] In certain embodiments, the compound is [ka] It is.

[0177] In certain embodiments, the compound is (E)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl)-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)-N-(((2R,3S,4S,5R)-3,4,5,6-tetrahydroxytetrahydro-2H-pyran-2-yl)methyl)acrylamide.

[0178] In certain embodiments, the compound is [ka] It is.

[0179] In certain embodiments, the compound is (E)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl)-N-(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)-N-(2,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acrylamide.

[0180] In certain embodiments, the compound is [ka] It is.

[0181] In certain embodiments, the compound is (E)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl)-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)-N-((3R,4R,5S,6R)-2,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acrylamide.

[0182] In certain embodiments, the compound is [ka] It is.

[0183] In certain embodiments, the compound is (E)-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. 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.

[0184] In certain embodiments, the compound is [ka] It is.

[0185] In certain embodiments, the compound is (E)-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. 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.

[0186] In certain embodiments, the compound is [ka] It is.

[0187] In certain embodiments, the compound is (E)-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. 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.

[0188] In certain embodiments, the compound is [ka] It is.

[0189] In certain embodiments, the compound is (E)-2-cyano-3-(6-(piperidin-1-yl)naphthalen-2-yl)-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)-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.

[0190] In certain embodiments, the compound is [ka] In certain embodiments, the compound is a pharma- ceutically acceptable salt or solvate of compound 21.

[0191] In certain embodiments, the compound is [ka] In certain embodiments, the compound is a pharma- ceutically acceptable salt or solvate of compound 22.

[0192] In certain embodiments, the compound is [ka] It is.

[0193] 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 pyrazidinyl. In certain embodiments, Ar2 is substituted or unsubstituted pyridyl.

[0194] 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 some cases, it is any atom or functional group that can donate part of its electron density to a conjugated pi-system through resonance or inductive electron withdrawal, thus 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 is heteroarylene, The alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene may optionally be one or more R 57 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 10Heterocycloalkyl, 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 , R 60 Each of the groups is independently hydrogen, C1 to 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 for hydrogen, is optionally represented by one or more R 61 is replaced by R 50 and R 51 are optionally joined together and optionally R 61 forming a heterocycloalkyl or heteroaryl substituted with R 61 Each of the groups is independently a halogen, -OR 62 , N.R. 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 the alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, arylene, or heteroarylene is optionally further represented by one or more R 65 is replaced by R 62 , R 63 and R 64 Each of is independently hydrogen or C1-C 10 alkyl, and each R 65 are independent, C1~C 10 Alkyl, C1-C 10Heteroalkyl, C1-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C1-C 10 Arylene, or C1-C 10 It is a heteroarylene.

[0195] 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.

[0196] 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.

[0197] 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.

[0198] 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.

[0199] In certain embodiments, x is 0, y is 0, z is 1, and Y is O.

[0200] In certain embodiments, x is 0, y is 0, z is 1, and Y is S.

[0201] In certain embodiments, x is 0, y is 0, z is 1, and Y is NH.

[0202] In certain embodiments, x is 0, y is 0, z is 1, and Y is absent.

[0203] In certain embodiments, x is 0, y is 0, z is 2, and Y is O.

[0204] In certain embodiments, x is 0, y is 0, z is 2, and Y is S.

[0205] In certain embodiments, x is 0, y is 0, z is 2, and Y is NH.

[0206] In certain embodiments, x is 0, y is 0, z is 2, and Y is absent.

[0207] In one aspect, the disclosure provides a compound of formula IIa: [ka] where EDG, Ar1, Ar2, Y, EWG, and WSG are defined above for Formula II.

[0208] In one aspect, the disclosure provides a compound of formula IIb: [ka] where EDG, Ar1, Ar2, EWG, and WSG are defined above for Formula II.

[0209] Optionally, the compound according to formula II is [ka] where n is an integer with a value from 0 to 50. In certain embodiments, n is an integer with a value from 0 to 10, e.g., n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0210] In certain embodiments, the compound is [ka] is selected from the group consisting of:

[0211] In certain embodiments, the compound is [ka] wherein R 85 is H or CN.

[0212] In certain embodiments, the compound is [ka] wherein R 85 is H or CN, and R 86 teeth [ka] where n is an integer with a value from 0 to 50. In certain embodiments, n is an integer with a value from 0 to 10, e.g., n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0213] In certain embodiments, the compound is [ka] wherein R 85 is H or CN, and R 86 is H.

[0214] In certain embodiments, the compound is [ka] wherein R 85 is H or CN, and R 86 teeth [ka] where n is an integer with a value from 0 to 50. In certain embodiments, n is an integer with a value from 0 to 10, e.g., n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0215] In certain embodiments, the compound is [ka] It is.

[0216] In certain embodiments, the compound is 2-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)-4-(6-piperidin-1-yl)naphthalen-2-yl)nicotinonitrile.

[0217] In certain embodiments, the compound is [ka] It is.

[0218] In certain embodiments, the compound is 4-(6-(piperidin-1-yl)naphthalen-2-yl)nicotinonitrile.

[0219] In certain embodiments, the compound is: [ka] or a salt or solvate thereof.

[0220] Treatment Methods and Uses Once a patient's traumatic brain injury (TBI) is determined, specific procedures can be provided to treat or ameliorate the symptoms of TBI or to reduce the possibility of further damage to the brain. Once TBI is diagnosed, disease progression can also be monitored by the methods described herein. Once diagnosed, the treating physician can also suggest additional treatments as described herein. In one embodiment, once TBI is detected or suspected, the patient is instructed to refrain from vigorous physical activity (e.g., sports, military service).

[0221] In some embodiments, the patient may be administered medication to treat or ameliorate TBI. Examples of treatments (medications) for TBI are provided below.

[0222] Drug Application Sedatives: These prevent excitement and excessive muscle activity and help with pain relief. Examples include Prophanol.

[0223] Painkillers: Opioids can be used.

[0224] Diuretics: These increase urine output and reduce water in the tissues. They are given intravenously. Mannitol is the diuretic most commonly used in TBI patients.

[0225] Anticonvulsant medications: People who experience moderate to severe TBI may have seizures for up to a week after the event. Medications can help prevent further brain damage that may result from seizures.

[0226] Coma-inducing drugs: During a coma, a person needs less oxygen. Sometimes, a coma can be intentionally induced if the blood vessels cannot supply enough food and oxygen to the brain.

[0227] surgery In some cases, surgery may be required.

[0228] Removal of hematomas: Internal bleeding can cause partially or fully clotted blood to pool in parts of the brain, worsening pressure on brain tissue. Emergency surgery can remove hematomas from between the skull and brain, reducing pressure within the skull and preventing further brain injury.

[0229] Repairing skull fractures: Any part of the skull that has broken and is pressing into the brain will need to be repaired surgically. Skull fractures that are not compressing the brain will usually heal on their own. The main concern with a skull fracture is that the force strong enough to cause it may have caused some underlying injury.

[0230] Creating an opening in the skull: This can relieve pressure within the skull if other interventions have not worked.

[0231] long term treatment People who have experienced severe TBI may need rehabilitation.

[0232] Depending on the extent and type of damage, a person may need to relearn how to walk, talk, and perform other daily tasks.

[0233] This may include treatment in a hospital or specialist treatment centre. Depending on the type of injury, physiotherapists, occupational therapists etc may be involved.

[0234] "Treatment" or "treating" is an approach to obtain 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 a disease or condition and / or reducing the severity of the disease or condition); b) improving, delaying or preventing the onset of one or more clinical symptoms associated with a disease or condition (e.g., stabilizing a disease or condition, preventing or delaying the worsening or progression of a disease or condition, and / or preventing or delaying the spread (e.g., metastasis) of a disease or condition; and / or c) relieving the disease, i.e., causing regression of clinical symptoms (e.g., improving the condition, providing partial or complete remission of a disease or condition, enhancing the effect of another drug, delaying disease progression, improving quality of life, and / or prolonging survival).

[0235] "Prevention" or "preventing" refers to any treatment of a disease or condition that does not result in the development of clinical symptoms of the disease or condition. The compounds, in some embodiments, may be administered to subjects (including humans) at risk for or who have a family history of the disease or condition.

[0236] "Subject" refers to an animal, such as a mammal (including a human), that has been or will be the object of treatment, observation or experiment. The methods described herein may be useful in human therapy and / or veterinary applications. In some embodiments, the subject is a mammal. In one embodiment, the subject is a human.

[0237] The term "therapeutically effective amount" or "effective amount" of a compound described herein or a pharma- ceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug, or deuterated analogue thereof means an amount sufficient to effect treatment when administered to a subject, to provide a therapeutic benefit, such as amelioration of symptoms or delay of disease progression. For example, a therapeutically effective amount may be an amount sufficient to reduce symptoms of a TBI disease or condition. A therapeutically effective amount may vary depending on the subject and the disease or condition being treated, the weight and age of the subject, the severity of the disease or condition, and the method of administration, but these can be readily determined by one of ordinary skill in the art.

[0238] The methods described herein can be applied to cell populations in vivo or ex vivo. "In vivo" means within a living individual, such as within an animal or human body. In this context, 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, including bodily 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 context, the compounds and compositions described herein can be used for a variety of purposes, including therapeutic and experimental purposes. For example, the compounds and compositions described herein can be used ex vivo to determine optimal schedules and / or dosing of administration of the disclosed compounds for a given indication, cell type, individual, and other parameters. Information gathered from such use can be used to set up in vivo treatment protocols, either for experimental purposes or in the clinic. Other ex vivo applications for which the compounds and compositions described herein may be suitable are described below or will become apparent to one of skill in the art. Selected compounds may be further characterized to determine safety or tolerated dosage in human or non-human subjects. Such properties may be determined using methods commonly known to those of skill in the art.

[0239] Administration and Pharmaceutical Compositions In some cases, the probe or compound is administered to the eye. In some cases, the pharmaceutical composition of the present disclosure administered to the eye is delivered to the retina, intraocular space, ocular surface, interconnecting nerve distribution, conjunctiva, lacrimal gland, or meibomian gland. In some cases, the compound is administered topically to the eye. In some cases, the compound is administered as an eye drop.

[0240] The probes or compounds can also be formulated for, but not limited to, intravenous and subcutaneous use. Intravenous administration can be by bolus or continuous infusion.

[0241] The probes are effective over a wide dosage range. In some cases, in adult applications, dosages of 0.01-1000 mg per day, 0.5-100 mg per day, 1-50 mg per day, and 5-40 mg per day are examples of dosages that may be used. An exemplary dosage is 10-30 mg per day. In youth applications, dosages may be the same as or less than adult dosages. In some cases, an effective amount of the probe corresponds to about 50-500 mg of the compound per adult subject. The exact dosage will depend on the route of administration, the form in which the compound is administered, the subject being treated, the body weight of the subject being treated, and the preference and experience of the attending physician.

[0242] In some cases, the effective amount of the probe corresponds to about 0.01-1000 mg of the compound per human subject per dose. In some cases, the effective amount of the compound is 50-500 mg per person per dose. In some cases, the effective amount is 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. ... 0mg, 0.1~700mg, 0.1~800mg, 0.1~900mg, 0.1~1000mg, 1~100mg, 1~200mg, 1~300mg, 1~400mg, 1~500mg, 1~600mg, 1~70 0mg, 1~800mg, 1~900mg, 100~200mg, 100~300mg, 100~400mg, 100~500mg, 100~600mg, 100~700mg, 100~800mg, 100~900m g, 100~1000mg, 200~300mg, 200~400mg, 200~500mg, 200~600mg, 200~700mg, 200~800mg, 200~900mg, 200~1000mg, 300 ~400mg, 300~500mg, 300~600mg, 300~700mg, 300~800mg, 300~900mg, 300~1000mg, 400~500mg, 400~600mg, 400~700mg, Equivalent to 400-800mg, 400-900mg, 400-1000mg, 500-600mg, 500-700mg, 500-800mg, 500-900mg, 500-1000mg, 600-700mg, 600-800mg, 600-900mg, 600-1000mg, 700-800mg, 700-900mg, 700-1000mg, 800-900mg, 800-1000mg, or approximately 900-1000mg.In some cases, the effective amount is 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.

[0243] 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 dosing is about once, twice, three times, four times, five times, six times, or more than six times per day. In some cases, the dosing is about once a month, once every two weeks, once a week, or once every two days. In other cases, the probe and another agent are administered together about once a day to about six times a day. In some cases, the administration of the probe and agent continues for less than about seven days. In yet other cases, the administration continues for about 6, 10, 14, 28 days, 2 months, 6 months, or a year or more. In some cases, continuous administration is achieved and maintained as long as necessary.

[0244] In some cases, the probe is administered 1-10 times, 1-4 times, or once per day. In some cases, the probe is administered 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times per day. In some cases, the probe is administered as droplets. In some cases, the size of the droplets administered ranges from 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 administers droplets in the range of about 10 to about 30 μL. An example of the present disclosure administers droplets in the range of about 10 to about 100 μL. An example of the present disclosure administers droplets in the range of about 20 to about 50 μL. An example of the present disclosure administers droplets in the range of about 20 to about 40 μL. An example of the present disclosure administers droplets in the range of about 10 to about 60 μL. In some cases, the ophthalmic formulations of the present disclosure are administered at a few drops per administration, for example, 1-3 drops per administration, 1-3 drops per administration, 1-4 drops per administration, 1-5 drops per administration, 1-6 drops per administration, 1-7 drops per administration, 1-8 drops per administration, 1-9 drops per administration, 1-10 drops per administration, 3-4 drops per administration, 3-5 drops per administration, 3-6 drops per administration, 3-7 drops per administration, 3-8 drops per administration, 3-9 drops per administration, 3-10 drops per administration, 5-6 drops per administration, 5-7 drops per administration, 5-8 drops per administration, 5-9 drops per administration, 5-10 drops per administration, 7-8 drops per administration, 7-9 drops per administration, or 9-10 drops per administration. In one example, the formulations of the present disclosure are administered at about 1 drop per administration and 1-6 times per day.

[0245] Pharmaceutical Compositions / Formulations In some cases, the probe described herein is formulated into pharmaceutical composition.In some cases, the pharmaceutical composition is formulated in a conventional manner using one or more physiologically acceptable carriers, including excipients and auxiliaries that facilitate the processing of active compound into pharmaceutically usable preparations.Suitable formulation depends on the route of administration selected. Any pharma- ceutically acceptable techniques, carriers, and excipients may be used as appropriate to formulate the pharmaceutical compositions described herein: Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999).

[0246] Provided herein is a pharmaceutical composition comprising the probe described herein and pharma- ceutically acceptable diluent(s), excipient(s) or carrier(s).In certain cases, the described compounds are administered as pharmaceutical compositions in which one or more probes are mixed with other active ingredients, such as in combination therapy.In specific cases, the pharmaceutical composition comprises one or more probes as described herein.

[0247] As used herein, a pharmaceutical composition refers to a mixture of any of the probe compounds described herein with other chemical components, such as carriers, stabilizers, diluents, dispersants, suspending agents, thickening agents, and / or excipients. In certain cases, the pharmaceutical composition facilitates administration of the compound to an organism. In some cases for carrying out the methods of treatment or use provided herein, a therapeutically effective amount of one or more of the probes provided herein is administered in a pharmaceutical composition to a mammal having a disease or condition to be detected, diagnosed or treated. In specific cases, the mammal is a human. In certain cases, the therapeutically effective amount varies depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used, and other factors. The compounds described herein are used as components of a mixture, alone or in combination with one or more therapeutic agents.

[0248] In some cases, one or more probes are formulated in an aqueous solution. In a specific case, the aqueous solution is selected from physiologically compatible buffers such as, by way of example only, Hanks' solution, Ringer's solution, acetate aqueous buffer, citrate aqueous buffer, carbonate aqueous buffer, phosphate aqueous buffer or saline buffer.

[0249] In some cases, the compounds described herein are formulated for ocular administration. In some cases, the ocular formulations are liquid (solutions, suspensions, powders for reconstitution, sol-to-gel systems), semi-solid (ointments and gels), solid (ocular inserts), and intraocular dosage forms (injections, irrigation solutions and implants).

[0250] Ophthalmic formulations comprising the compounds described herein and ophthalmologically acceptable components are provided herein.Ophthalmic formulations can be administered in any form suitable for ocular administration, such as solutions, suspensions, ointments, gels, liposomal dispersions, colloidal microparticle suspensions, etc., or in ocular inserts, such as optionally biodegradable controlled release polymer matrices.Importantly, at least one component of the formulation, preferably two or more formulation components, are "multifunctional" in that they are useful for preventing or treating multiple conditions and disorders, or have two or more mechanisms of action, or both.

[0251] By "pharmacologically acceptable" or "ophthalmically acceptable" is meant an ingredient that is not biologically or otherwise undesirable, i.e., the ingredient may be incorporated into the ophthalmic formulations of the present invention and administered topically to the eye of a patient without causing undesirable biological effects or interacting in a deleterious manner with any of the other ingredients of the formulation composition in which it is included. When the term "pharmacologically acceptable" is used to refer to an ingredient other than a pharmacologically active drug, it means that the ingredient has met the necessary standards of toxicological and manufacturing testing or that it is included in the Inactive Ingredient Guide prepared by the US Food and Drug Administration.

[0252] The formulation also includes an effective amount of a penetration enhancer that enhances the penetration of the formulation components through cell membranes, tissues, and extracellular matrices, including the cornea. An "effective amount" of a penetration enhancer refers to 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, by way of example, methylsulfonylmethane (MSM; also known as methylsulfone), a combination of MSM and dimethylsulfoxide (DMSO), or in less preferred embodiments, a combination of MSM and DMSO, with MSM being particularly preferred.

[0253] Kits and Packages Also provided herein are kits and packages that include the probes of the present disclosure, retinal imaging devices, and optionally suitable packaging. In one embodiment, the kit further includes instructions for use.

[0254] The retinal imaging device may include a lens(es) and an image sensor for detecting the signal emitted from the probe used (thus forming a suitable retinal scanner). In some embodiments, the retinal imaging device detects a fluorescent signal. In some embodiments, the retinal imaging device further includes a laser light source that can be used to activate the fluorescent signal from the probe. EXAMPLES

[0255] The following examples are included to illustrate specific embodiments of the present disclosure.Those skilled in the art should understand that the techniques disclosed in the following examples represent techniques that work well in implementing the disclosure, and therefore can be considered to constitute specific modes for its implementation.However, those skilled in the art should understand in light of the present disclosure that many changes can be made in the specific embodiments disclosed and still obtain similar or similar results without departing from the spirit and scope of the disclosure.

[0256] Example 1: Ex vivo analysis of probes using human AD tissue Postmortem retinal tissue and brains from humans with confirmed or probable Alzheimer's disease (AD) were obtained and examined for amyloid beta (Aβ) by staining with a test amyloid-sensitive fluorescent probe (ASF), Compound 1. This example was performed to determine whether ASF can identify patients with amyloid burden in the brain through fluorescent examination of ASF-stained retinas.

[0257] As shown in Figure 1, dense deposits measuring 5–10 μm in diameter could be fluorescently visualized in flat-mounted retinal samples stained with ASF. In this example, these deposits could be observed in both AD (panels A–B) and cognitively normal patients (panels C–D), with a qualitative correlation between the density of these deposits in the retina and the extent of amyloidosis in the brain (estimated by immunohistochemistry (IHC) using anti-Aβ antibodies).

[0258] IHC staining of retinal cryosections provided further evidence of the presence of Aβ in retinal deposits with similar location, size, and morphology to deposits seen in ASF-stained retinal flat-mount sections, providing promising evidence that ASF can bind to amyloid deposits in ex vivo human retinal tissue.

[0259] Example 2: Visual acuity testing using TBI blast mouse model This example was performed to determine whether ASF Compound 1 can detect Aβ in the retina of a TBI mouse model following injury.

[0260] Retinal tissues from blast mouse model and uninjured mice 24 hours after injury were stained with ASF and co-stained with anti-Aβ antibody (6E10). As shown in Figure 2, mice subjected to blast injury expressed immunoreactivity to Aβ in retinal tissues where uninjured mice did not show reactivity to 6E10. More importantly, ASF fluorescently labels these retinal deposits (Figure 2, top row), but uninjured mice do not show any fluorescence enhancement (Figure 2, bottom row).

[0261] This provides evidence that ASF probe compound 1 has the ability to detect retinal amyloid present in TBI mouse models. Based on this evidence, this example further sought to confirm whether Aβ could also be detected in brain tissue of the same TBI blast mouse model. As shown in Figure 3, mice subjected to blast injury displayed immunoreactivity to Aβ in brain tissue where uninjured mice showed no reactivity to 6E10. The evidence of Aβ in brain tissue complements that found in retinal tissue of the same injured mice. It also supports previous studies on the correlation between brain and retina in Aβ accumulation.

[0262] Example 3: Controlled Cortical Impact in C5BL / 6 Mice Three-month-old C5BL / 6 mice were briefly anesthetized with isoflurane and positioned in a stereotaxic frame. Mice underwent craniotomy over the right primary and secondary motor cortex and parieto-temporal cortex (+1 to -4 anterior-posterior from bregma, 4 mm lateral from the sagittal suture). Half of the mice were not further injured to serve as control specimens. In the remaining mice, a 3 mm diameter piston was positioned in the center of the motor cortex approximately +0.5 to -2.5 mm from bregma and 3 mm lateral to the sagittal suture. A stereotaxic impactor (Impact One, myNeuroLab.com) was used to accelerate the piston at a speed of 3 m / s to an impact depth of 1 mm below the cortical surface. After 24 h, mice were euthanized and brains and eyes were harvested. Retinas were extracted and stained with DAPI, Compound 1, and 6E10 antibodies for comparison.

[0263] The retinas were mounted on slides and washed twice with phosphate-buffered saline (PBS) for 5 min each time. The retinas were exposed to 98% formic acid for 1 min and then washed twice with distilled water for 5 min each time. The retinas were then equilibrated in 1× PBS for 15 min. The retinas were then washed twice with 1× PBS for 15 min. The retinas were then equilibrated in ... (登録商標)The retina was exposed to 10% goat / donkey serum in 1× phosphate buffered saline (PBST) containing 20 for 1 hour. The retina was then incubated with 610 antibody in 10% goat serum in 1× PBST overnight at 4°C. The retina was then rinsed 3 times with 1× PBST for 5 minutes each time. The retina was then incubated with the secondary antibody Alexa Fluor568 anti-mouse in 1× PBST for 1 hour while blocking light. The retina was rinsed 3 times with 1× PBST for 5 minutes each time. The retina was then stained with compound 1 working solution at room temperature for 30 minutes while blocking light. The retina was then washed 3 times with 1× PBS for 5 minutes each time, followed by staining with 300 nM or 100 ng / mL of DAPI for 10 minutes in the dark. The retina was then washed 3 times with PBS for 10 minutes each time. DAKO mounting medium was applied and the samples were kept out of light until imaging.

[0264] 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 the fluorescent probes associated with DAPI (blue), compound 1 (green), and 6E10 antibody (red): 408, 488, and 568 nm. Z-stack images were taken at 40x with 0.5 μm increments to visualize the entire thickness of the tissue.

[0265] Figures 4-7 show that after CCI, 3-month-old C5BL / 6 mice developed deposits of aggregated proteins that were detected by compound 1 and 6E10 antibodies in the retina. The aggregated proteins were detected by 6E10 in the retina and colocalized with compound 1.

[0266] Example 4: Detection of retinal Aβ aggregation in an ex vivo mouse model This example was performed to examine whether TBI induced retinal Aβ aggregation in a mouse model.

[0267] Mice underwent controlled cortical impact (CCI). The CCI model involved the application of a controlled impact to the intact dura after craniotomy, which is a commonly used TBI animal model. Mice were anesthetized, their heads were mounted in a stereotaxic frame, and a craniotomy was performed over the right side of the frontal cortex. Using a stereotaxic impactor, mice were impacted over the right side of the frontal cortex with a piston of 3 m / s velocity, 1 mm depth, and 3 mm diameter. 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 consisted of exposure to anesthesia, stereotaxy, skin and fascia reflexion, and closure of the incision with staples. Mice were then euthanized 24 h after injury, and retinal tissue was dissected for further analysis.

[0268] The mouse retinas were extracted and stained with compound 1 together with 6E10 antibody, a sequence-specific antibody for Aβ. As shown in FIG. 8, mice that underwent CCI showed immunoreactivity for Aβ along retinal blood vessels where intact mice showed no reactivity for 6E10. More importantly, compound 1 fluorescently labeled these deposits and colocalized with 6E10 immunoreactivity (FIG. 8, top, white arrows), but did not show fluorescence enhancement in intact mice (FIG. 8, bottom). This example shows that the compounds of the present disclosure can effectively detect retinal amyloid in a TBI mouse model.

[0269] Example 4: Detection of retinal Aβ aggregation in an ex vivo mouse model This example was performed to examine whether TBI induced retinal Aβ aggregation in a mouse model.

[0270] Mice underwent controlled cortical impact (CCI). The CCI model involved the application of a controlled impact to the intact dura after craniotomy, which is a commonly used TBI animal model. Mice were anesthetized, their heads were mounted in a stereotaxic frame, and a craniotomy was performed over the right side of the frontal cortex. Using a stereotaxic impactor, mice were impacted over the right side of the frontal cortex with a piston of 3 m / s velocity, 1 mm depth, and 3 mm diameter. 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 consisted of exposure to anesthesia, stereotaxy, skin and fascia reflexion, and closure of the incision with staples. Mice were then euthanized 24 h after injury, and retinal tissue was dissected for further analysis.

[0271] The mouse retinas were extracted and stained with compound 1 together with 6E10 antibody, a sequence-specific antibody for Aβ. As shown in FIG. 8, mice that underwent CCI showed immunoreactivity for Aβ along retinal blood vessels where intact mice showed no reactivity for 6E10. More importantly, compound 1 fluorescently labeled these deposits and colocalized with 6E10 immunoreactivity (FIG. 8, top, white arrows), but did not show fluorescence enhancement in intact mice (FIG. 8, bottom). This example shows that the compounds of the present disclosure can effectively detect retinal amyloid in a TBI mouse model.

[0272] Example 5: Synthesis Synthesis of compound 21 [ka] Synthesis of (6-bromonaphthalen-2-yl)methanol (25) To a solution of LiAlH4 (8.2 g, 217 mmol in 500 mL THF) at 0° C., under N2, was added dropwise a solution of methyl 6-bromo-2-naphthoate (24) (50.0 g, 189 mmol) in 500 mL anhydrous THF. The reaction mixture was left stirring at 0° C. for 1 h. Once the reaction was complete as monitored by TLC, the mixture was treated with H2O, 15% NaOH, H2O (1:1:3, v / v / v). After filtration, the filtrate was concentrated, extracted with EA and dried over NaSO4. The crude product was purified from (PE:EA=3:1) to give the title compound.

[0273] Synthesis of 6-bromo-2-naphthaldehyde (26) To a suspension of (6-bromonaphthalen-2-yl)methanol (25) (42.0 g, 177 mmol) and silica gel (76.4 g) in DCM (500 mL) was added PCC (76.4 g, 354 mmol). The reaction was stirred at RT (room temperature, 25±5° C.) for 1.5 h. Upon completion, it was filtered through a pad of silica and concentrated under reduced pressure to give the title compound.

[0274] Synthesis of 6-(piperidin-1-yl)-2-naphthaldehyde (27) To dry and degassed toluene (300 mL) was added 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). The reaction was left stirring at 115 °C for 8 h. After cooling, the mixture was filtered, washed with EA, then concentrated to one third of the volume, to which was added 200 ml of 6N hydrochloric acid with thorough stirring. The aqueous phase was separated and extracted 3 times with DCM, then made alkaline with 5N NaOH and extracted with EA. The organic phase was concentrated to give the crude material, which was further purified by silica gel chromatography (PE:EA=20:1 to 2:1) to give the title compound.

[0275] Synthesis of 2-cyano-N-(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)acetamide (29) In a pear-shaped flask, 28 (6.0 g, 40 mmol) was added to methyl 2-cyanoacetate (4.0 g, 40 mmol) with stirring. The mixture was stirred at room temperature overnight and concentrated to give the crude material, which was used directly in the next step.

[0276] Synthesis of (E)-2-cyano-N-(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)-3-(6-(piperidin-1-yl)naphthalen-2-yl)acrylamide (Compound 21) 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 h. The reaction was then concentrated under reduced pressure to give the crude material which was purified by silica gel chromatography to give the title compound. Exact mass 437.23; m / e 437.23 (100%), 438.23 (28.9%), 439.24 (4.7%); elemental analysis (C 25 H 31 N3O4): C 68.63%, H 7.14%, N 9.60%, O 14.63%.

[0277] Synthesis of compound 22 [ka] Synthesis of 31 To 1 g (2.3 mM) of compound 21 in 15 mL of THF at zero temperature was added 1.6 g (3 mM) of tetrabenzyl pyrophosphate prepared using Merck Organic Synthesis preparation to give a deep red solution. Sodium hydride (100 mg, 2.5 mM, 60% in oil) was added. After 15 min warming to room temperature solids started to precipitate. DMF (5 mL) was added and stirred at room temperature for 1 h. Water (200 mL) and ethyl acetate (200 mL) were added. The ethyl acetate layer was dried and evaporated. Purification by ISCO® on an 80 g silica gel cartridge using 0-100% hexane / ethyl acetate gave the title compound. Compound 8 1 The H NMR spectrum is presented in Figures 1A-1C. MS (m / z) 701.3 [M+H] + .

[0278] Synthesis of compound 22 To 1 gram of (compound 31) was added argon degassed 95% ethanol (150 mL). 120 mg of 10% Pd / C was added and hydrogen gas was bubbled through the reaction mixture for 5 minutes. The mixture was then stirred under a hydrogen balloon for 3 hours. The reaction mixture was degassed with argon and evaporated. Purification was carried out by preparative LC / MS on a 25X250mm C18 column using 0-100% water containing 2g / L ammonium acetate. After free drying, compound 22 was obtained. 1 The H NMR spectrum (DO) is presented in Figures 2A-B.

[0279] Synthesis of compound 23 [ka] A solution of compound 31 (5 mmol, 3.49 g, 1 equiv.) in anhydrous CH2Cl2 (100 ml) was cooled to 0 °C under argon and trimethylsilyl bromide (50 mmol, 6.8 ml, 10 equiv.) was added via syringe. The reaction mixture was cooled to 0 oThe mixture was stirred at RT for 30 min and the completion of conversion was monitored by HPLC. The mixture was then quenched with MeOH (ca. 50 mL) and stirred for 10 min. The solution was evaporated to dryness. This process was repeated four times.

[0280] The organic solvent was evaporated under vacuum, the residue was suspended in a trace of MeOH, and EtOAc was added to induce precipitation of the phosphonic acid. The residue was filtered and washed with EtOAc (×2). The residue was then dried under vacuum to give the desired phosphate.

[0281] The phosphonic acid was treated with NHOAc (25 mmol, 1.93 g, 5 eq.) and 150 ml water at room temperature and stirring was continued for an additional 15 min resulting in 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).

[0282] Example 6: Detection of retinal Aβ aggregation in vivo Following the explicit ex vivo experiments in Example 4, in vivo retinal imaging studies were performed using mice before and after undergoing CCI.

[0283] To obtain baseline retinal images, mice were imaged before undergoing TBI with CCI. Anesthetized mice received intravenous administration of compound 23 (15 mg / kg of a 20 mg / mL solution in 0.1 M phosphate buffer, pH 7.4) before CCI and 24 hours after CCI. As shown in Figure 9, background fluorescence enhancement was observed in the retinal vasculature 3 minutes after administration of compound 23 in intact mice. This enhancement was no longer observed 15 minutes after administration of compound 23 (Figure 9, top).

[0284] Twenty-four hours after baseline imaging, mice were anesthetized, their heads mounted in a stereotaxic frame, and a craniotomy was performed over the right side of the motor cortex. Using a stereotaxic impactor, mice were impacted over the right side of the motor cortex using a piston with a speed of 5 m / s, a depth of 2 mm, and a diameter of 3 mm. After injury, the incision was closed with stitches, anesthesia was terminated, and the animals were placed on a heated pad to maintain normal core temperature. 24 hours after CCI, live retinal imaging was performed in anesthetized mice after intravenous administration of compound 23. As shown in Figure 9, the retinal vasculature remained illuminated throughout the 15-minute imaging period, indicating that Aβ may be present within the blood vessels (Figure 9, bottom row). This is in contrast to pre-CCI imaging, where vascular fluorescence enhancement was visible 3 minutes after injection but decreased over the 15-minute imaging period.

[0285] This example shows that within 24 hours after CCI, compound 23 can be used to detect changes in the retinal vasculature that may indicate Aβ accumulation. This example is consistent with the ex vivo data shown in Figure 8, where amyloid accumulation appeared along the blood vessels of mice 24 hours after CCI. ***

[0286] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0287] The present invention illustratively described herein can suitably be practiced in the absence of any element(s), limitation(s) not specifically disclosed herein. Thus, for example, terms such as "comprising," "including," "containing," etc., should be read broadly and without limitation. The terms and expressions used are used as terms of description and not of limitation, and it is not intended that the use of such terms and expressions excludes any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the claims.

[0288] Thus, while the present invention has been specifically disclosed by preferred embodiments and optional features, modifications, improvements and variations of the invention embodied in the present invention disclosed herein may occur to those skilled in the art, and such modifications, improvements and variations are considered to be within the scope of the present invention. The materials, methods, and examples provided herein are representative of preferred embodiments, are illustrative, and are not intended to limit the scope of the invention.

[0289] The present invention has been described broadly and generically herein. Each of the narrower species and subgeneric groupings falling within the generic disclosure also form part of the present invention. This includes the generic description of the invention with a provisos or negative limitation removing the subject matter from the genus, regardless of whether the removed material is specifically described herein.

[0290] Additionally, where features or aspects of the disclosure are described in terms of Markush groups and other groupings of alternatives, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual members or subgroups of members of the Markush group or other group.

[0291] 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 was individually incorporated by reference. In case of conflict, the present specification, including definitions, will control.

[0292] While the present disclosure has been described in conjunction with the above embodiments, it should be understood that the foregoing description and examples are intended to be illustrative, and not limiting, of the scope of the present disclosure. Other aspects, advantages, and modifications within the scope of the present disclosure will be apparent to those skilled in the art to which the present disclosure pertains.

Claims

[Claim 1] The invention described in the specification.

Images