Methods, compositions and systems for evaluating visual function

Abstract

Provided herein is a method of treating a subject having an ocular disease. The method comprises administering a treatment to the subject. The method further includes determining whether the treatment is therapeutically effective for the subject based on the composite biomarker. The composite biomarker is calculated based on at least two variables. At least one of the at least two variables is associated with one or more selected from the group consisting of vision, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof.

Description

[0001] Cross - reference to related applications

[0002] This application claims the benefit of U.S. Application No. 63 / 331,189, filed Apr. 14, 2022, which is incorporated herein by reference in its entirety. BACKGROUND OF THE INVENTION

[0003] There is a need for effective clinical assessment of disease progression and treatment response in eye diseases and disorders such as Stargardt disease (STGD) and age-related macular degeneration (AMD). SUMMARY OF THE INVENTION

[0004] Disclosed herein are methods and systems for treating a subject having an eye disease. Disclosed herein is a method for treating a subject having an eye disease, the method comprising: (a) administering a treatment to the subject; (b) determining whether the treatment is therapeutically effective for treating the subject based on a composite biomarker, wherein the composite biomarker is calculated based on at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof.

[0005] Disclosed herein is a method for treating a subject having an eye disease, the method comprising: (a) administering a treatment to the subject; (b) assaying the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (c) calculating the at least two variables to produce a composite biomarker; and (d) determining whether the treatment is therapeutically effective for treating the subject based on the composite biomarker.

[0006] Disclosed herein is a method for treating a subject having an eye disease, the method comprising: (a) administering a treatment to the subject; (b) performing a first assay on the subject at a first time point to obtain a first set of at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (c) calculating the at least two variables of the first set to produce a first composite biomarker; (d) performing a second assay on the subject at a second time point to obtain a second set of the at least two variables; (e) calculating the at least two variables of the second set to produce a second composite biomarker; and (f) determining whether the treatment is therapeutically effective for treating the subject based on the first and second composite biomarkers.

[0007] The present disclosure provides methods for treating a subject having an eye disease, the methods comprising: (a) administering a treatment to the subject; (b) measuring the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, eye anatomy, eye physiology, eye pathology, and changes thereof; and (c) calculating the at least two variables to produce a composite biomarker, wherein when the composite biomarker exceeds a threshold, the treatment is therapeutically effective for treating the subject, and wherein when the composite biomarker does not exceed the threshold, the treatment is not therapeutically effective for treating the subject.

[0008] The present disclosure provides methods for treating a subject having an eye disease, the methods comprising: (a) administering a first dose of a treatment to the subject; (b) measuring the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, eye anatomy, eye physiology, eye pathology, and changes thereof; (c) calculating the at least two variables to produce a composite biomarker; (d) determining whether the first dose of the treatment is therapeutically effective for treating the subject based on the composite biomarker; and (e) administering a second dose of the treatment to the subject, wherein when it is determined that the first dose is not therapeutically effective for treating the subject, the second dose is different from the first dose, and wherein when it is determined that the first dose is therapeutically effective for treating the subject, the second dose is substantially the same as the first dose.

[0009] The present disclosure provides methods for treating a subject having an eye disease, the methods comprising: (a) performing a first measurement on the subject to obtain a first set of at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, eye anatomy, eye physiology, eye pathology, and changes thereof; (b) calculating the at least two variables of the first set to produce a first composite biomarker; (c) administering a treatment to the subject; (d) performing a second measurement on the subject to obtain a second set of the at least two variables; (e) calculating the at least two variables of the second set to produce a second composite biomarker; and (f) determining whether the treatment is therapeutically effective for treating the subject based on the first and second composite biomarkers.

[0010] The present disclosure provides methods for treating a subject having an eye disease, the methods comprising: (a) measuring the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, eye anatomy, eye physiology, eye pathology, and changes thereof; (b) calculating the at least two variables to produce a composite biomarker; (c) selecting a treatment for the subject based on the composite biomarker; and (d) administering the treatment to the subject.

[0011] The present disclosure provides methods for treating a subject having an eye disease, the methods comprising: (a) performing a first determination on the subject to obtain at least two variables of a first set, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) calculating the at least two variables of the first set to produce a first composite biomarker; (c) selecting a treatment for the subject based on the first composite biomarker; (d) administering the treatment to the subject; (e) performing a second determination on the subject to obtain the at least two variables of a second set; (f) calculating the at least two variables of the second set to produce a second composite biomarker; and (g) determining whether the treatment is therapeutically effective for treating the subject based on the first and second composite biomarkers.

[0012] The present disclosure provides methods for treating a subject having an eye disease, the methods comprising: (a) performing a determination on the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) calculating the at least two variables to produce a composite biomarker; (c) selecting a treatment for the subject when the composite biomarker exceeds a threshold, or optionally, not selecting the treatment for the subject when the composite biomarker does not exceed the threshold; and (d) administering the treatment to the subject when the treatment is selected for the subject, or optionally, not administering the treatment to the subject when the treatment is not selected for the subject.

[0013] The present disclosure provides methods for treating a subject having an eye disease, the methods comprising: (a) performing a first determination on the subject to obtain at least two variables of a first set, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) calculating the at least two variables of the first set to produce a first composite biomarker; (c) selecting a treatment for the subject based on the first composite biomarker; (d) administering a first dose of the treatment to the subject; (e) performing a second determination on the subject to obtain the at least two variables of a second set; (f) calculating the at least two variables of the second set to produce a second composite biomarker; (g) determining whether the first dose of the treatment is therapeutically effective for treating the subject based on the first and second composite biomarkers; and (h) administering a second dose of the treatment to the subject, wherein the second dose is different from the first dose when it is determined that the first dose is not therapeutically effective for treating the subject, and the second dose is substantially the same as the first dose when it is determined that the first dose is therapeutically effective for treating the subject.

[0014] The present disclosure provides methods for treating a subject having an eye disease, the methods comprising: (a) performing a first measurement on the subject to obtain at least two variables of a first set, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, eye anatomy, eye physiology, eye pathology, and changes thereof; (b) calculating the at least two variables of the first set to produce a first composite biomarker; (d) administering a first dose of a treatment to the subject; (e) performing a second measurement on the subject to obtain the at least two variables of a second set; (f) calculating the at least two variables of the second set to produce a second composite biomarker; (g) determining that the first dose of the treatment is therapeutically effective for treating the subject when the difference between the first and second composite biomarkers exceeds a threshold, or optionally, determining that the first dose of the treatment is not therapeutically effective for treating the subject when the difference between the first and second composite biomarkers does not exceed the threshold; and (h) administering a second dose of the treatment to the subject, wherein the second dose is different from the first dose when it is determined that the first dose is not therapeutically effective for treating the subject, and wherein the second dose is substantially the same as the first dose when it is determined that the first dose is therapeutically effective for treating the subject.

[0015] The present disclosure provides methods for monitoring the outcome of a treatment for an eye disease in a subject, the methods comprising determining whether a dose of the treatment administered to the subject is therapeutically effective for treating the subject by calculating at least two variables of the subject to produce a composite biomarker, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, eye anatomy, eye physiology, eye pathology, and changes thereof.

[0016] The present disclosure provides methods for treating a subject having an eye disease, which comprise administering a therapeutically effective dose of a treatment to the subject, wherein the treatment is selected for the subject based on a composite biomarker, wherein the composite biomarker is calculated based on at least two variables of the subject, and wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, eye anatomy, eye physiology, eye pathology, and changes thereof.

[0017] The present disclosure provides methods, which comprise selecting a treatment for a subject having an eye disease based on a composite biomarker, wherein the composite biomarker is calculated based on at least two variables of the subject, and wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, eye anatomy, eye structure, eye physiology, eye pathology, and changes thereof.

[0018] The present disclosure provides a method for selecting a treatment for a subject having an eye disease, the method comprising: (a) calculating at least two variables of the subject to produce a composite biomarker, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; and (b) selecting the treatment for the subject when the composite biomarker exceeds a threshold, or optionally, not selecting the treatment for the subject when the composite biomarker does not exceed the threshold.

[0019] The present disclosure provides a method for selecting a treatment for a subject having an eye disease, the method comprising: (a) assaying the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) calculating the at least two variables to produce a composite biomarker; and (c) selecting the treatment for the subject when the composite biomarker exceeds a threshold, or optionally, not selecting the treatment for the subject when the composite biomarker does not exceed the threshold.

[0020] The present disclosure provides a method for predicting a treatment response of a subject to a treatment for an eye disease, the method comprising: (a) calculating at least two variables of the subject to produce a composite biomarker, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; and (b) predicting the treatment response of the subject to the treatment based on the composite biomarker.

[0021] The present disclosure provides a method for predicting a treatment response of a subject to a treatment for an eye disease, the method comprising: (a) assaying the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) calculating the at least two variables to produce a composite biomarker; and (c) determining that the subject is likely to have a treatment response to the treatment when the composite biomarker exceeds a threshold, or optionally, determining that the subject is likely to not have a treatment response to the treatment when the composite biomarker does not exceed the threshold.

[0022] The present disclosure provides a method for determining a dosage of a treatment for a subject having an eye disease based on a composite biomarker, wherein the composite biomarker is calculated based on at least two variables of the subject, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof.

[0023] The present disclosure provides a method for determining a dosage for treating an ocular disease in a subject, the method comprising: (a) measuring the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) calculating the at least two variables to generate a composite biomarker; and (c) determining the dosage for the treatment of the subject based on the composite biomarker.

[0024] The present disclosure provides a method for determining a dosage for treating an ocular disease in a subject, the method comprising: (a) performing a first measurement on the subject to obtain at least two variables of a first set, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) calculating the at least two variables of the first set to generate a first composite biomarker; (b) administering the treatment to the subject; (b) performing a second measurement on the subject to obtain at least two variables of a second set; (c) calculating the at least two variables of the second set to generate a second composite biomarker; and (d) determining the dosage for the treatment of the subject based on the first and second composite biomarkers.

[0025] The present disclosure provides a method comprising administering a second dosage of a treatment to a subject having an ocular disease, wherein based on a composite biomarker calculated from at least two variables of the subject, it is determined that a first dosage of the treatment previously administered to the subject is lacking in therapeutic efficacy for treating the ocular disease of the subject, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof, and wherein the second dosage is different from the first dosage.

[0026] The present disclosure provides a method for determining a dosage for treating an ocular disease in a subject, the method comprising: (a) administering a dosage of the treatment to the subject; (b) measuring the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (c) calculating the at least two variables to generate a composite biomarker; (d) determining whether the dosage of the treatment is therapeutically effective for treating the subject having the ocular disease based on the composite biomarker.

[0027] The present disclosure provides a method for determining a dosage for treating an object suffering from an eye disease, the method comprising: (a) performing a first measurement on the object to obtain at least two variables of a first set, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) calculating the at least two variables of the first set to generate a first composite biomarker; (c) administering a dosage of the treatment to the object; (d) performing a second measurement on the object to obtain the at least two variables of a second set; (e) calculating the at least two variables of the second set to generate a second composite biomarker; and (f) determining that the dosage of the treatment is therapeutically effective for treating the object when the difference between the first and second composite biomarkers exceeds a threshold, or optionally, determining that the dosage of the treatment is not therapeutically effective for treating the object when the difference between the first and second composite biomarkers does not exceed the threshold.

[0028] In some embodiments, the ocular disease includes macular lesions, retinopathy, retinal atrophy, macular atrophy, macular degeneration, age-related macular degeneration (AMD), Stargardt disease (STGD), RP (retinitis pigmentosa), ABCA4 gene mutations, or combinations thereof. In some embodiments, a sample from the subject having the ocular disease has an expression level of retinol binding protein 4 (RBP4) that is at least a threshold. In some embodiments, the threshold is about 25 μg / ml. In some embodiments, the threshold is about 35 μg / ml. In some embodiments, the threshold is from about 25 μg / ml to about 100 μg / ml. In some embodiments, the expression level of RBP4 is measured by an assay including an antibody assay, an electrophoresis assay, an immunoassay, a radioimmunoassay, a chromatography assay, a mass spectrometry assay, a microarray-based detection assay, a polymerase chain reaction assay, a sequencing assay, an immunohistochemistry assay, or any combination thereof. In some embodiments, the antibody assay includes ELISA. In some embodiments, a sample from the subject having the ocular disease has an expression level of vitamin A that is at least a threshold. In some embodiments, the presence or absence of one or more genomic variants indicates that the subject has the ocular disease. In some embodiments, the presence or absence of one or more genomic variants indicates that the subject has Stargardt disease. In some embodiments, the threshold is about 150 ng / mL. In some embodiments, the threshold is about 225 ng / mL or about 390 ng / mL. In some embodiments, the threshold is from about 150 ng / mL to about 500 ng / mL. In some embodiments, the expression level of the vitamin is measured by an assay including an antibody assay, an electrophoresis assay, an immunoassay, a radioimmunoassay, a chromatography assay, a mass spectrometry assay, a microarray-based detection assay, a polymerase chain reaction assay, a sequencing assay, an immunohistochemistry assay, or any combination thereof. In some embodiments, the sample includes a blood sample. In some embodiments, the expression level of the RBP4 or vitamin A is measured from plasma or serum derived from the blood sample. In some embodiments, the presence or absence of one or more genomic variants selected from rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, and rs1801574 indicates that the subject has the ocular disease. In some embodiments, the one or more genomic variants include at least 4 of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574.In some embodiments, the one or more genomic variants include at least 5 of rs3747961, rs6666652, rs1800717, rs763108716, rs185601596, rs17110761, rs61748519, rs1801359, rs145766145, rs76258939, rs200551567, rs754765164, rs201602424, rs564661476, rs4147831, rs6657239, rs2297632, rs1801555, rs1762114, rs55860151, rs1800549, rs3112831, rs4147830, rs2297634, or rs4847281.

[0029] In some embodiments, the treatment includes a pharmaceutical composition that comprises an RBP4 inhibitor or a compound configured to reduce the blood RBP4 concentration of the subject. In some embodiments, the pharmaceutical composition comprises a compound having the structure of formula (I):

[0030]

[0031] Wherein: R A 1 , R A 2 , R A 3 , R A 4 and R A 5 are each independently H, halogen, CF 3 or C 1 -C 4 alkyl, wherein two or more of R A 1 , R A 2 , R A 3 , R A 4 and R A 5 are not H; R A 6 is H, OH or halogen; and AA has the structure:

[0032]

[0033] Wherein α, β, χ and δ are each independently absent or present, and when present are each a bond; X is C or N; Z 1 is N; Z2 is N or NR A 9 wherein R A 9 is H, C 1 -C 4 alkyl or oxetane; B A is a substituted or unsubstituted 5-, 6- or 7-membered ring structure; or a pharmaceutically acceptable salt thereof.

[0034] In some embodiments, the compound has the structure or a pharmaceutically acceptable salt thereof. In some embodiments, the compound has the structure or a pharmaceutically acceptable salt thereof.

[0035] In some embodiments, the pharmaceutical composition comprises a compound having the structure of formula (II):

[0036]

[0037] wherein: Ring A B is optionally further substituted benzene; R B 1 is optionally substituted branched C 3 -C 6 alkyl; X B 1 is O, S, SO, SO 2 or NH; X B 2 is a bond or C 1 -C 3 alkylene; Ring B B is azetidine or piperidine; X B 3 is CO or SO 2 ; R B 2 is an optionally substituted hydrocarbyl, an optionally substituted heterocyclyl, an optionally substituted hydroxy, an optionally substituted mercapto, cyano, nitro, acyl or halogen atom; or a pharmaceutically acceptable salt thereof.

[0038] In some embodiments, the compound is 4-(3-(2-tert-butylphenoxy)azetidin-1-yl)-4-oxobutanoic acid, 3-{3-[(2-tert-butyl-4-fluorophenoxy)methyl]azetidin-1-yl}-3-oxopropanoic acid, 2-{[3-(2-tert-butyl-4-chlorophenoxy)azetidin-1-yl]carbonyl}pyridine, 4-[3-(2-tert-butyl-4-chlorophenoxy)azetidin-1-yl]-4-oxobutanoic acid, {3-[(2-tert-butyl-4-chlorophenoxy)methyl]azetidin-1-yl}(oxo)acetic acid, {3-[(2-tert-butylphenoxy)methyl]azetidin-1-yl}(oxo)acetic acid, 3-{3-[(2-tert-butylphenoxy)methyl]azetidin-1-yl}-3-oxopropanoic acid, {4-[(2-tert-butyl-4-chlorophenoxy)methyl]piperidin-1-yl}(oxo)acetic acid, [4-(2-tert-butylphenoxy)piperidin-1-yl](oxo)acetic acid or {4-[(2-tert-butylphenoxy)methyl]piperidin-1-yl}(oxo)acetic acid, or a pharmaceutically acceptable salt thereof.

[0039] In some embodiments, the pharmaceutical composition comprises a compound having the structure of formula (III):

[0040]

[0041] Wherein: Ring A C is a benzene ring optionally substituted with 1 to 3 substituents selected from: (a) a halogen atom and (b) C 1 -C 6 alkyl; Ring B C is a piperazine ring optionally substituted with 1 to 3 substituents selected from: (a) a halogen atom, (b) C 1 -C 6 alkyl optionally substituted with 1 to 3 halogen atoms and (c) C 1 -C 6 alkoxy optionally substituted with 1 to 3 halogen atoms; and R C is (1) optionally substituted C 1 -C 10 alkyl, (2) optionally substituted C 6 -C 14 aryl, (3) optionally substituted 5- or 6-membered aromatic heterocyclic group, (4) optionally substituted amino group, (5) optionally substituted carboxyl group or (6) optionally substituted carbamoyl group, or a pharmaceutically acceptable salt thereof.

[0042] In some embodiments, the compound is N-{[4-(2-tert-butylphenyl)piperazin-1-yl]carbonyl}glycine, 3-[4-(2-tert-butylphenyl)piperazin-1-yl]-3-oxopropanoic acid, [4-(2-tert-butyl-4-chlorophenyl)piperazin-1-yl](oxo)acetic acid, 5-{2-[4-(2-tert-butylphenyl)piperazin-1-yl]-2-oxoethyl}imidazolidine-2,4-dione, [(5-{[4-(2-tert-butylphenyl)piperazin-1-yl]carbonyl}isoxazol-3-yl)oxy]acetic acid, or a pharmaceutically acceptable salt thereof.

[0043] In some embodiments, the pharmaceutical composition comprises a compound having the structure of formula (IV):

[0044]

[0045] Wherein: Ring A D is a 5-membered non-aromatic heterocycle, optionally further substituted by an oxo group; Ring B D is a benzene ring, which is optionally further substituted by 1 to 4 substituents; and X D is O, CH 2 O, OCH 2 、CH 2 、(CH 2 ) 2 、S、CH 2 S、SCH 2 、S(O)、CH 2 S(O)、S(O)CH 2 、S(O) 2 、CH 2 S(O) 2 or S(O) 2 CH 2 ; or a pharmaceutically acceptable salt thereof.

[0046] In some embodiments, the compound is ((3S)-1-[3,5-bis(trifluoromethyl)phenyl]pyrrolidin-3-yl)oxy)acetic acid, ((1-[4-chloro-3-(trifluoromethyl)phenyl]pyrrolidin-3-yl)thio)acetic acid, 3-((2R,5S)-5-[3,5-bis(trifluoromethyl)phenyl]tetrahydrofuran-2-yl)propanoic acid, or a pharmaceutically acceptable salt thereof.

[0047] In some embodiments, the pharmaceutical composition comprises a compound having the structure of formula (V):

[0048]

[0049] Wherein, Ring A E is a pyrazole ring, a pyridine ring, an oxazole ring, an imidazole ring or a pyrimidine ring; XE is S, optionally substituted alkylene or O; and R E is a hydrogen atom or C 1 -C 6 alkyl; or a pharmaceutically acceptable salt thereof.

[0050] In some embodiments, the compound is ((4-(3,5-bis(trifluoromethyl)phenyl)-1,3-oxazol-2-yl)thio)acetic acid, ethyl ((6-(3,5-bis(trifluoromethyl)phenyl)pyridin-3-yl)thio)acetate, ((6-(3,5-bis(trifluoromethyl)phenyl)pyridin-3-yl)thio)acetic acid or 3-(3-(3,5-bis(trifluoromethyl)phenyl)-1H-pyrazol-1-yl)butyric acid or 3-{3-[3,5-bis(trifluoromethyl)phenyl]-1H-pyrazol-1-yl}propionic acid (also known as STG-001).

[0051] In some embodiments, at least one of the at least two variables is related to one or more selected from visual acuity, the average score of a functional vision questionnaire, and measurements obtained from an imaging technique. In some embodiments, at least one of the at least two variables is related to one or more selected from changes in visual acuity, changes in the average score of a functional vision questionnaire, and changes in measurements obtained from an imaging technique for detecting or quantifying changes in ocular structure or visual function. In some embodiments, at least one of the at least two variables is related to changes in visual acuity, changes in the average score of a functional vision questionnaire, and changes in measurements obtained from an imaging technique for detecting or quantifying changes in optic nerve atrophy or visual function. In some embodiments, the at least two variables are related to changes in visual acuity, changes in the average score of a functional vision questionnaire, and changes in measurements obtained from an imaging technique for detecting or quantifying changes in optic nerve atrophy or visual function. In some embodiments, the at least two variables are related to changes in visual acuity and changes in the average score of a functional vision questionnaire. In some embodiments, the at least two variables are related to changes in visual acuity and changes in measurements obtained from an imaging technique for detecting or quantifying changes in optic nerve atrophy or visual function. In some embodiments, the at least two variables are related to changes in the average score of a functional vision questionnaire and changes in measurements obtained from an imaging technique for detecting or quantifying changes in optic nerve atrophy or visual function. In some embodiments, both of the at least two variables are related to changes in visual acuity. In some embodiments, both of the at least two variables are related to changes in the average score of a functional vision questionnaire. In some embodiments, both of the at least two variables are related to changes in measurements obtained from an imaging technique for detecting or quantifying changes in optic nerve atrophy or visual function. In some embodiments, at least one of the at least two variables is related to changes in ocular structure, changes in ocular pathology, changes in retinal atrophy, changes in macular atrophy, changes in macular degeneration, or a combination thereof. In some embodiments, two of the at least two variables are related to each other. In some embodiments, one of the at least two variables is related to visual acuity. In some embodiments, one of the at least two variables is related to changes in visual acuity. In some embodiments, all of the at least two variables are related to changes in visual acuity. In some embodiments, one of the at least two variables is related to the status of the ocular disease. In some embodiments, one of the at least two variables is related to the progression of the ocular disease. In some embodiments, the at least two variables include values obtained from a logarithm of the minimum angle of resolution (logMAR) chart, a Snellen chart, a best corrected visual acuity (BCVA) test, an Early Treatment Diabetic Retinopathy Study (ETDRS) letter test, or a combination thereof.In some embodiments, the at least two variables include values obtained from a functional vision questionnaire, which preferably includes one or more questions related to: (a) the subject's confidence in moving to different places during the day or at night; (b) the subject's ability to recognize people, participate in sports, or obtain information; and (c) the subject's ability to engage in social interactions or work efficiency, where each of the one or more questions is scored for the subject. In some embodiments, each of the one or more questions is scored for the subject on a severity scale of 0 - 4 as follows: (a) an answer of "never" is scored 0; (b) an answer of "almost never" is scored 1; (c) an answer of "sometimes" is scored 2; (d) an answer of "almost always" is scored 3; and (e) an answer of "always" is scored 4. In some embodiments, the at least two variables include values obtained from microperimetry (MP), perimetry, or both. In some embodiments, the MP includes a spatial map measuring the retinal sensitivity of the subject. In some embodiments, the MP includes measuring the average retinal sensitivity of the subject or its variation. In some embodiments, the at least two variables include values obtained from one or more selected from fundus photography, fundus angiography (FA), fundus autofluorescence (FAF), spectral domain - optical coherence tomography (SD - OCT), quantitative autofluorescence (qAF), infrared (IR) imaging, optical coherence tomography - angiography (OCT - A), and wide - field imaging. In some embodiments, the at least two variables include one or more selected from the area of questionable decreased autofluorescence (QDAF), the area of definite decreased autofluorescence (DDAF), the area of decreased autofluorescence (DAF), retinal thickness, ellipsoid zone (EZ) defect width, central subfield retina thickness (CST), outer subfield retina thickness (OST), middle subfield retina thickness (MST), and their variations. In some embodiments, the area of decreased autofluorescence (DAF) includes the area of questionable decreased autofluorescence (QDAF) and the area of definite decreased autofluorescence (DDAF). In some embodiments, the at least two variables include two or more selected from visual acuity, QDAF, DAF, EZ defect width, or their variations.In some embodiments, the at least two variables include visual acuity, QDAF, DAF, EZ defect width, or a variation thereof. In some embodiments, the EZ defect width is related to the visual acuity. In some embodiments, the EZ defect width is related to the QDAF. In some embodiments, the EZ defect width is related to the DAF. In some embodiments, the visual acuity is related to the QDAF. In some embodiments, the EZ defect width is related to the visual acuity, QDAF, and DAF. In some embodiments, the visual acuity is related to the EZ defect width and QDAF. In some embodiments, the EZ defect width is related to the visual acuity, QDAF, and DAF, with a correlation coefficient greater than about 0.8 and a p-value less than about 0.05. In some embodiments, the visual acuity is related to the EZ defect width and QDAF, with a p-value less than about 0.05. In some embodiments, the at least two variables of the subject are obtained from the right and left eyes of the subject. In some embodiments, the method further includes predicting whether the prognostic potential of a treatment is improved based on one or more composite biomarkers. In some embodiments, when the composite biomarker exceeds the threshold, the treatment is therapeutically effective for treating the subject. In some embodiments, when the composite biomarker does not exceed the threshold, the treatment is not therapeutically effective for treating the subject.

[0052] Incorporated by reference

[0053] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference for the specific purposes described herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the invention will be obtained by reference to the following detailed description that illustrates illustrative embodiments that make use of the principles of the invention and the accompanying drawings, in which:

[0055] Figure 1A and Figure 1B illustrate the reduction in autofluorescence (AF) of qAF in the right eye ( Figure 1A ) and left eye ( Figure 1B ) of Subject 3 after treatment with an RBP4 inhibitor. The left image in each panel is labeled Phase 2 Baseline, and the right image in each panel is labeled Month 6, Phase 2. The qDAF values for each image are labeled (left to right): 335, 275, 351, and 268.

[0056] Figure 1C illustrate fundus autofluorescence (FAF) of the right eye of Subject 3 at Phase 2 Baseline (left) and Month 6, Phase 2 (right). The lesion area is labeled 2.7 mm2 (Left) and 2.7 mm 2 (Right).

[0057] Figure 1D The figure shows the EZ defect analysis of the left eye of subject 3 at the second stage baseline (left) and at the sixth month of the second stage (right). The red lines are marked as 2.91 mm (left) and 2.71 mm (right).

[0058] Figure 1E The figure shows the changes in MST and OST (retinal thickness) of the left eye of subject 3 after treatment with an RBP4 inhibitor. The top image is marked as the second stage baseline, and the bottom image is marked as the sixth month of the second stage. The left side of each image shows the area of the eye and the average thickness. For the second stage baseline, the average thickness values in the legend are (from top to bottom): marked (97 microns); central (97 microns); central minimum (97 microns); and central maximum (202 microns). For the sixth month of the second stage, the average thickness values in the legend are (from top to bottom): marked (112 microns); central (112 microns); central minimum (104 microns); and central maximum (216 microns). The right side of each image shows the area of the eye and the retinal thickness; the retinal thickness scale on the right is marked from 0 to 800 at 100-unit intervals; the scale bar inserted in the image is marked as 200 microns.

[0059] Figure 2A The figure shows the stabilization of the QDAF size in the left eye of subject 4 after treatment with an RBP4 inhibitor. The left image is marked as the second stage baseline, and the right image is marked as the sixth month of the second stage.

[0060] Figure 2B The figure shows the changes in MST and OST (retinal thickness) of the left eye of subject 4 after treatment with an RBP4 inhibitor. The top image is marked as the second stage baseline, and the bottom image is marked as the sixth month of the second stage. The left side of each image shows the area of the eye and the average thickness. For the second stage baseline, the average thickness values in the legend are (from top to bottom): marked (60 microns); central (63 microns); central minimum (57 microns); and central maximum (218 microns). For the sixth month of the second stage, the average thickness values in the legend are (from top to bottom): marked (56 microns); central (58 microns); central minimum (54 microns); and central maximum (199 microns). The right side of each image shows the area of the eye and the retinal thickness; the retinal thickness scale on the right is marked from 0 to 800 at 100-unit intervals; the scale bar inserted in the image is marked as 200 microns.

[0061] Figure 2CIllustrated is the EZ defect analysis of the right eye of subject 4 at the 2nd stage baseline (left) and at the 6th month of the 2nd stage (right). The red lines are marked as 1.79 mm (left) and 1.85 mm (right).

[0062] Figure 3A Illustrated are the changes in SD-OCT in the right eye of subject 11 after treatment with an RBP4 inhibitor. The left figure is marked as the 2nd stage baseline, and the right figure is marked as the 6th month of the 2nd stage.

[0063] Figure 3B Illustrated are the changes in autofluorescence in the right eye of subject 11 after treatment with an RBP4 inhibitor. The left figure is marked as the 2nd stage baseline, and the right figure is marked as the 6th month of the 2nd stage.

[0064] Figure 3C Illustrated is the EZ defect analysis of the left eye of subject 11 at the 2nd stage baseline (left) and at the 6th month of the 2nd stage (right). The red lines are marked as 2.20 mm (left) and 2.29 mm (right). Detailed Description

[0065] As used herein and in the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a reagent" includes a plurality of such reagents, and reference to "the cell" includes reference to one or more cells (or a plurality of cells) and equivalents thereof known to those skilled in the art, and so forth. When ranges are used herein for physical properties (such as molecular weight) or chemical properties (such as chemical formula), it is intended to include all combinations and sub-combinations of the ranges and specific embodiments therein. When referring to a numerical value or a numerical range, the term "about" means that the numerical value or numerical range referred to is an approximation within experimental variability (or within statistical experimental error), and thus in some cases, the numerical value or numerical range will vary between 1% and 15% of the stated numerical value or numerical range. The term "comprising" (and related terms such as "having" or "including") is not intended to exclude "consisting of the described features" or "consisting essentially of the described features" in other certain embodiments (such as embodiments of any combination of substances, compositions, methods, or processes, etc. described herein).

[0066] Ocular Diseases and Treatments

[0067] The present disclosure provides methods for assessing disease progression and disease treatment of an ocular disease of an object. The ocular disease includes a disease that affects visual function, ocular structure, ocular anatomy, retinal structure, retinal function, retinal pathology, macular structure, macular function, macular pathology, or a combination thereof. For example, the ocular disease includes but is not limited to macular lesions, retinopathy, retinal atrophy, macular atrophy, macular degeneration, diabetic retinopathy, inherited retinal diseases, age-related macular degeneration (AMD), Stargardt disease (STGD), RP (retinitis pigmentosa), ABCA4 gene mutations, or a combination thereof. Patients with retinal degenerative diseases may experience loss or decrease of visual acuity (VA) or pathological or structural changes in the retina. Age-related macular degeneration (AMD) is a common ocular condition in populations over 50 years old and is also a major cause of vision loss. It damages the macula, a small spot near the center of the retina and the part of the eye required for sharp central vision. As AMD progresses, it is common to have a blurred area near the center of vision. Over time, the blurred area may become larger, and the object may develop a blind spot in their central vision. Stargardt disease (STGD) is a juvenile macular lesion with a pathology similar to AMD. STGD is the more common form of inherited macular degeneration, and STGD1 is the most common subtype. One in 10,000 people globally has STGD. Early-onset Stargardt disease (<20 years old) is usually associated with a more severe condition. In retinal degenerative diseases including STGD and AMD, the accumulation of lipofuscin has been observed, which is related to tissue autofluorescence (AF) in the retinal pigment epithelium (RPE), and bisretinoid N-retinyl-N-retinylidene ethanolamine (A2E) is a component of lipofuscin. The accumulation of A2E may be the cause of retinal atrophy. The excessive accumulation of lipofuscin has been recognized as a common pathogenic pathway for various retinal diseases including AMD, preceding photoreceptor degeneration. The appearance of AF is related to disease pathology. Quantitative AF can be a direct assessment of the relative levels of A2E and related bisretinoids in retinal tissue. Studies have shown that the AF level of childhood-onset STGD1 is significantly higher than that of adult-onset STGD1. The AF of childhood-onset STGD1 is 8 times higher than that of healthy eyes. Studies have shown that childhood-onset STGD1 is characterized by a rapid decline in visual acuity. Studies have shown that adult-onset STGD1 is a milder phenotype with foveal preservation, which keeps visual acuity stable for several years. Mutations in ATP-binding cassette subfamily A member 4 (ABCA4) may lead to the excessive accumulation of the cytotoxic bisretinoid A2E. A2E is the main source of abnormal autofluorescence (AF) in the retina. The continuous increase in AF in STGD1 and AMD may lead to retinal atrophy and subsequent photoreceptor loss, resulting in a decrease in AF.These retinopathies are referred to as areas of definite AF reduction (DDAF) and can be quantified using imaging techniques. See Tanna, P. et al. (2016) British Journal of Ophthalmology, 101(1):25-30; Invest Ophthalmol Vis Sci. 2011;52:9379–9390; Invest Ophthalmol Vis Sci. 2015;56:8179-8186; Invest Ophthalmol Vis Sci. 2014;55:2841–2852; Ophthalmology 2015;122:335-344; Br J Ophthalmol 2017;101:25-30; AM J Ophthalmol 2020;211:159-175; JAMA Ophthalmol 2017;135:1232-1241; Invest Ophthalmol Vis Sci. 2016;57:5186–5191. Retinol binding protein 4 (RBP4) is a retinol transport protein in the blood, secreted by adipocytes and the liver. Reducing RBP4 levels can lead to a decrease in lipofuscin accumulation, which causes vision loss in diseases such as AMD, dry (atrophic) AMD, and STGD. Reducing RBP4 can reduce lipofuscin accumulation in the retina, thereby delaying or preventing vision loss due to excessive lipofuscin accumulation in the retina in AMD or STGD. The RBP4 inhibitors disclosed herein can inhibit RBP4 to slow down bisretinoid / A2E accumulation.

[0068] Visual function assessment

[0069] There are various methods for evaluating eye diseases or visual function for various purposes, including but not limited to selecting treatment for an object, evaluating the disease progression of an object, evaluating the potential therapeutic effect of a treatment on an object, evaluating the therapeutic or clinical response of an object to a treatment, determining the dose of a treatment for an object, or a combination thereof. These methods include but are not limited to retinal imaging methods, imaging modalities, and functional tests. Imaging modalities typically use a dedicated camera with / without non-invasive lasers or fluorescent dyes. Imaging modalities include but are not limited to fundus photography, fundus angiography (FA), fundus autofluorescence (FAF), spectral domain-optical coherence tomography (SD-OCT), quantitative autofluorescence (qAF), infrared (IR) imaging, optical coherence tomography-angiography (OCT-A), and wide-field imaging. Functional tests typically evaluate visual function or the impact on daily activities or behaviors. Functional tests include visual acuity tests, functional vision questionnaires, microperimetry, visual field tests, and other behavioral function tests. These evaluation methods are used to measure many variables related to visual acuity, visual function, visual structure, ocular anatomy, retinal atrophy, retinal degeneration, macular degeneration, retinal physiology or pathology, macular physiology or pathology, or changes thereof. For example, these variables include but are not limited to visual acuity (VA), functional vision questionnaire score, logarithm of the minimum angle of resolution (logMAR) chart score, Snellen chart score, best corrected visual acuity (BCVA) test score, Early Treatment Diabetic Retinopathy Study (ETDRS) letter test score, retinal sensitivity, size of questionable decreased autofluorescence (QDAF), size of definite decreased autofluorescence (DDAF), size of decreased autofluorescence (DAF), retinal thickness, ellipsoid zone (EZ) defect width, central subfield retinal thickness (CST), outer subfield retinal thickness (OST), middle subfield retinal thickness (MST), or changes thereof, or any other variable related to visual function.

[0070] Methods, compositions, and systems for assessing visual function are disclosed herein. In some embodiments, the methods, compositions, or systems disclosed herein can combine at least two variables to generate a composite biomarker. In some embodiments, the composite biomarker is used to assess an eye disease or visual function. In some embodiments, the composite biomarker is used to select a treatment for a subject with an eye disease, evaluate the progression of the subject's eye disease, evaluate the potential therapeutic effect of a treatment on a subject with an eye disease, evaluate the treatment response of a subject to a treatment for an eye disease, determine a dose of a treatment for a subject, or any other purpose related to treating or evaluating an eye disease, or a combination thereof. In some embodiments, the at least two variables are obtained from at least two assessment methods for eye diseases disclosed herein, including but not limited to imaging methods, imaging modalities, or functional tests. In some embodiments, the at least two variables include measurements, images, analyses, test scores, values, or any other form of data obtained from an eye disease assessment method. In some embodiments, the composite biomarker can be in the form of a score, a predictive score, a predictive model, or a combination thereof. In some embodiments, the composite biomarker includes a threshold. In some embodiments, whether the composite biomarker exceeds the threshold is related to a certain outcome of the methods disclosed herein. In some embodiments, the composite biomarker is used to select a subject for a certain clinical treatment. In some embodiments, the composite biomarker is used to evaluate the progression of a subject's disease. In some embodiments, the composite biomarker is used to evaluate the potential therapeutic effect of a certain treatment on a subject before treatment. In some embodiments, the composite biomarker is used to determine a dose of a treatment for a subject. In some embodiments, the composite biomarker is used to evaluate the therapeutic effect of a treatment on a subject during treatment. In some embodiments, the composite biomarker is used for a combination of multiple purposes disclosed herein. In some embodiments, when the composite biomarker exceeds the threshold, a certain clinical treatment can be selected for the subject. In some embodiments, when the composite biomarker does not exceed the threshold, a certain clinical treatment cannot be selected for the subject. In some embodiments, when the composite biomarker exceeds the threshold, it can be determined that the subject may have a certain disease state. In some embodiments, when the composite biomarker does not exceed the threshold, it can be determined that the subject is less likely to have a certain disease state. In some embodiments, when the composite biomarker exceeds the threshold, it can be determined that the subject may benefit from a treatment before the treatment is administered. In some embodiments, when the composite biomarker does not exceed the threshold, it can be determined that the subject is less likely to benefit from a treatment before the treatment is administered. In some embodiments, when the composite biomarker exceeds the threshold, it can be determined that the subject may have a certain treatment response to the treatment during the treatment process. In some embodiments, when the composite biomarker does not exceed the threshold, it can be determined that the subject is less likely to have a certain treatment response to the treatment during the treatment process.In some embodiments, a dose for the treatment of an eye disease can be calculated based on a composite biomarker.

[0071] In some embodiments, at least one of the at least two variables is related to the state of the eye disease. In some embodiments, the at least two variables include at least one objective variable. In some embodiments, at least one objective variable is related to the state of the eye disease. In some embodiments, at least one objective variable includes a change in the eye structure. In some embodiments, at least one objective variable includes an eye pathological change. In some embodiments, at least one of the at least two variables is related to another of the at least two variables. In some embodiments, each of the at least two variables is related to the remaining variables of the at least two variables. In some embodiments, at least one of the at least two variables is related to visual acuity. In some embodiments, visual acuity can be an endpoint of clinical assessment or clinical treatment. In some embodiments, visual acuity can be a subjective measurement. In some embodiments, the at least two variables include visual acuity, a visual questionnaire score, a measurement obtained from an imaging technique used to evaluate the eye disease, a change thereof, or a combination thereof. In some embodiments, the imaging technique is used to detect and / or quantify a change in retinal atrophy. In some embodiments, the at least two variables include a change in visual acuity, a change in the average visual questionnaire score, a measurement from an imaging technique used to detect and / or quantify a change in retinal atrophy. In some embodiments, one of the at least two variables is obtained from the right eye, left eye, or both eyes of the subject. In some embodiments, the at least two variables are obtained from at least two subjects having different genders, races, ages, or a combination thereof. In some embodiments, the at least two variables are obtained during at least two clinical stages.

[0072] There are many characteristic variables available for evaluating an eye disease, including but not limited to a change in visual acuity, a change in the average score of a functional visual questionnaire, a change in a measurement obtained from an imaging modality used to detect and / or quantify a change in the structure / anatomy of the retina or macula (such as retinal atrophy, macular atrophy, or macular degeneration in AMD and STGD). These characteristic variables are used to select a treatment for a patient, monitor disease progression, and / or evaluate treatment efficacy.

[0073] In some embodiments, visual acuity can be evaluated using a logarithm of the minimum angle of resolution chart (logMAR chart). The logMAR chart includes letter rows for estimating visual acuity. Zero logMAR can represent standard visual acuity, a positive value can represent poorer visual acuity, and a negative value can represent better visual acuity. In some embodiments, the Early Treatment Diabetic Retinopathy Study (ETDRS) letter score for best corrected visual acuity (BCVA) can be calculated using the following formula:

[0074] LogMAR score = -0.02 * letter score + 1.7

[0075] In some embodiments, a modifier of -1 can be added to the change in visual acuity to account for improved visual acuity. In some embodiments, a Snellen chart can also be used to evaluate visual acuity.

[0076] In some embodiments, a functional vision questionnaire can be conducted among multiple questions scored on a severity scale. An average score is obtained from one or more functional vision questionnaires. In some embodiments, the scoring scale of the vision questionnaire can be any range (or including the two numbers) between two numbers. In some embodiments, the scoring scale can be from 0 to 4. In some embodiments, a score is assigned to the answer "never" to a question according to the scoring scale. In some embodiments, a score is assigned to the answer "almost never" to a question according to the scoring scale. In some embodiments, a score is assigned to the answer "sometimes" to a question according to the scoring scale. In some embodiments, a score is assigned to the answer "almost always" to a question according to the scoring scale. In some embodiments, a score is assigned to the answer "always" to a question according to the scoring scale. In some embodiments, according to the 0-4 scoring scale, scores of 0, 1, 2, 3, and 4 are respectively assigned to the answers "never", "almost never", "sometimes", "almost always", and "always" to a question. In some embodiments, according to the 0-4 scoring scale, scores of 4, 3, 2, 1, and 0 are respectively assigned to the answers "never", "almost never", "sometimes", "almost always", and "always" to a question.

[0077] In some embodiments, the functional vision questionnaire includes any number of questions. In some embodiments, the functional vision questionnaire includes any number of questions from about 1 to about 200, such as about 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, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199 or about 200, or any number therebetween. In some embodiments, the functional vision questionnaire contains about 24 questions.

[0078] In some embodiments, the questions in the visual questionnaire relate to visual function. In some embodiments, visual function includes the confidence of an object in moving to different places during the day. In some embodiments, visual function includes the confidence of an object in moving to different places at night. In some embodiments, visual function includes the confidence of an object in moving to different places during the day and at night. In some embodiments, visual function includes the confidence of an object in moving to different places during the day or at night. In some embodiments, visual function includes the ability of an object to recognize people. In some embodiments, visual function includes the ability of an object to participate in sports. In some embodiments, visual function includes the ability of an object to obtain information. In some embodiments, visual function includes the ability of an object to recognize people, participate in sports, or obtain information. In some embodiments, visual function includes the social interaction of an object. In some embodiments, visual function includes the work efficiency of an object. In some embodiments, visual function includes the social interaction and work efficiency of an object. In some embodiments, visual function includes the social interaction or work efficiency of an object.

[0079] Fundus autofluorescence (FAF) imaging is used as an endpoint in clinical trials. FAF is used to detect lesion growth by measurements at multiple time points. The change in the lesion area measured by FAF can be plotted against time to calculate the lesion growth rate, e.g., how fast or slow the macula or retina of a patient is degenerating compared to a placebo. FAF imaging is used to compare the rate of anatomical or structural deterioration or change to evaluate the therapeutic effect of a drug or treatment, e.g., reducing the deterioration of retinal degeneration.

[0080] In some embodiments, fundus autofluorescence (FAF) is used to evaluate eye diseases or visual function. In some embodiments, one or more retinal images of FAF are used to detect areas of atrophy. In some embodiments, one or more retinal images of FAF are used to gain a detailed understanding of the health of the retinal pigment epithelium (RPE). In some embodiments, one or more images of FAF can be high-contrast retinal images. In some embodiments, two types of decreased autofluorescence (DAF) can be quantified. In some embodiments, one type of DAF can be defined DAF (DDAF), which includes areas with a darkness level defined as qualitatively determined. In some embodiments, the other type of DAF can be questionable DAF (QDAF), which includes areas with a darkness level defined as qualitatively questionable. In some embodiments, the size of the decreased autofluorescence (DAF) lesion can be the sum of all DDAF and QDAF. In some embodiments, DAF can be expressed as an area in square millimeters (mm 2 )

[0081] In some embodiments, spectral domain - optical coherence tomography (SD - OCT) is used to evaluate eye diseases or visual function. In some embodiments, SD - OCT provides in - vivo cross - sectional and surface imaging of the retina and the retinal pigment epithelium (RPE). In some embodiments, SD - OCT is used to detect the presence of RPE atrophy. In some embodiments, SD - OCT is used to evaluate the width of the ellipsoid zone (EZ) defect. In some embodiments, the EZ line provides information about photoreceptor integrity. In some embodiments, the EZ line provides useful information for early diseases. In some embodiments, the images of SD - OCT are used to evaluate the central subfield retinal thickness (CST). In some embodiments, the images of SD - OCT are used to evaluate the mid - peripheral subfield retinal thickness (MST). In some embodiments, the images of SD - OCT are used to evaluate the outer subfield retinal thickness (OST).

[0082] In some embodiments, microperimetry is used to evaluate eye diseases or visual function. In some embodiments, microperimetry is used to measure the mean retinal sensitivity. In some embodiments, the retinal sensitivity can be the response level of the retina to light stimulation. In some embodiments, microperimetry is used to spatially map the retinal sensitivity. In some embodiments, microperimetry can be a psychophysical visual function test. In some embodiments, microperimetry is used to identify the correlation between anatomical features and visual function.

[0083] In some embodiments, a method for measuring quantitative autofluorescence (qAF) is used to evaluate eye diseases or visual function. In some embodiments, the qAF method can incorporate an internal fluorescence reference to account for variable laser power and detector sensitivity. In some embodiments, the intensity of AF can be calculated based on a calibrated internal fluorescence reference standard in the optical path. Methods for evaluating visual function using composite biomarkers

[0084] In one aspect, the present disclosure provides a method of treating a subject having an eye disease. In some embodiments, the method comprises one or more steps disclosed herein. In some embodiments, a method comprises one or more steps of different methods disclosed herein. In some embodiments, one or more steps of the methods disclosed herein may be combined with one or more steps of another method disclosed herein. In some embodiments, the method comprises administering to the subject a treatment for the eye disease. In some embodiments, the treatment comprises a pharmaceutical composition disclosed herein. In some embodiments, the treatment may be administered at a dose disclosed herein. In some embodiments, the method comprises obtaining at least two variables by assaying the subject. In some embodiments, at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, and ocular pathology. In some embodiments, at least one of the at least two variables is related to one or more selected from a change in visual acuity, a change in visual function, a change in ocular anatomy, a change in ocular physiology, and a change in ocular pathology. In some embodiments, the assay comprises at least two assays. In some embodiments, the at least two assays may be performed at two or more time points to obtain at least two variables. In some embodiments, the at least two assays may be performed at a first time point and a second time point. In some embodiments, the assay comprises any number of assays. In some embodiments, the assay comprises any number of assays in the range of 1 to 100, such as 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, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100, or any number therebetween.In some embodiments, the number of time points at which measurements are made includes any number in the range of 1 to 100, such as 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, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100, or any number therebetween. In some embodiments, at least one of the at least two variables is a variable disclosed herein. In some embodiments, the at least two variables include all of the variables disclosed herein. In some embodiments, the method includes the step of calculating at least two variables to produce a composite biomarker. In some embodiments, the composite biomarker can be calculated based on at least two variables. In some embodiments, the composite biomarker can be calculated based on a formula or model disclosed herein. In some embodiments, the method includes the step of determining whether a treatment is therapeutically effective for a subject to be treated based on the composite biomarker. In some embodiments, the treatment is therapeutically effective for the subject to be treated when the composite biomarker exceeds a threshold. In some embodiments, the treatment is not therapeutically effective for the subject to be treated when the composite biomarker does not exceed the threshold.

[0085] In some embodiments, a method of treating a subject having an eye disease includes, after the step of administering a treatment, making a first determination of the subject at a first time point to obtain at least two variables of a first set. In some embodiments, the method includes calculating the at least two variables of the first set to generate a first composite biomarker. In some embodiments, the first composite biomarker can be calculated based on the at least two variables of the first set. In some embodiments, the method includes making a second determination of the subject at a second time point to obtain at least two variables of a second set. In some embodiments, the method includes calculating the at least two variables of the second set to generate a second composite biomarker. In some embodiments, the second composite biomarker can be calculated based on the at least two variables of the second set. In some embodiments, the method includes determining whether the treatment is therapeutically effective for the treated subject based on the first and second composite biomarkers. In some embodiments, the method includes calculating composite biomarkers based on the at least two variables of the first set and the second set. In some embodiments, the method includes determining whether the treatment is therapeutically effective for the treated subject based on the composite biomarkers calculated from the at least two variables of the first set and the second set. In some embodiments, the composite biomarker can be calculated from the at least two variables of the first set and the second set based on the formulas or models disclosed herein. In some embodiments, the method includes comparing the first and second composite biomarkers to obtain a difference between the first and second composite biomarkers. In some embodiments, when the difference between the first and second composite biomarkers exceeds a threshold, the treatment is therapeutically effective for the treated subject. In some embodiments, when the difference between the first and second composite biomarkers does not exceed a threshold, the treatment is not therapeutically effective for the treated subject.

[0086] In some embodiments, a method of treating a subject having an eye disease includes the step of administering a first dose of a treatment to the subject. In some embodiments, the method includes the step of assaying the subject after administering the first dose of the treatment to obtain at least two variables. In some embodiments, the method includes the step of calculating the at least two variables to produce a composite biomarker. In some embodiments, the composite biomarker can be calculated based on the at least two variables. In some embodiments, the method includes the step of determining whether the first dose of the treatment is therapeutically effective for treating the subject based on the composite biomarker. In some embodiments, the method includes the step of administering a second dose of the treatment to the subject. In some embodiments, the second dose can be different from the first dose when it is determined that the first dose is not therapeutically effective for treating the subject. In some embodiments, the second dose can be the same as or substantially the same as the first dose when it is determined that the first dose is therapeutically effective for treating the subject. In some embodiments, the first dose of the treatment is therapeutically effective for treating the subject when the composite biomarker exceeds a threshold. In some embodiments, the first dose of the treatment is not therapeutically effective for treating the subject when the composite biomarker does not exceed the threshold.

[0087] In some embodiments, the method includes, after the step of administering a first dose of a treatment, performing a first determination on the subject at a first time point to obtain at least two variables of a first set. In some embodiments, the method includes performing a second determination on the subject at a second time point to obtain at least two variables of a second set. In some embodiments, a first composite biomarker can be calculated based on at least two variables of the first set. In some embodiments, a second composite biomarker can be calculated based on at least two variables of the second set. In some embodiments, the method includes determining whether the treatment is effective for the treated subject based on the first and second composite biomarkers. In some embodiments, the method includes calculating a composite biomarker based on at least two variables of the first set and the second set. In some embodiments, the method includes determining whether the treatment is effective for the treated subject based on the composite biomarker calculated from at least two variables of the first set and the second set. In some embodiments, the composite biomarker can be calculated from at least two variables of the first set and the second set based on the formulas or models disclosed herein. In some embodiments, the method includes the step of administering a second dose of the treatment to the subject. In some embodiments, the second dose can be different from the first dose when it is determined that the first dose is not effective for the treated subject. In some embodiments, the second dose can be the same as or substantially the same as the first dose when it is determined that the first dose is effective for the treated subject. In some embodiments, the method includes comparing the first and second composite biomarkers to obtain a difference between the first and second composite biomarkers. In some embodiments, the treatment is effective for the treated subject when the difference between the first and second composite biomarkers exceeds a threshold. In some embodiments, the treatment is not effective for the treated subject when the difference between the first and second composite biomarkers does not exceed a threshold. In some embodiments, the treatment is effective for the treated subject when the composite biomarker calculated from at least two variables of the first set and the second set exceeds a threshold. In some embodiments, the treatment is not effective for the treated subject when the composite biomarker calculated from at least two variables of the first set and the second set does not exceed a threshold.

[0088] In some embodiments, a method of treating a subject having an eye disease includes, prior to the step of administering a treatment, making a first determination of the subject at a first time point to obtain at least two variables of the first set disclosed herein. In some embodiments, the method includes, prior to the step of administering a treatment, calculating at least two variables of the first set to produce a first composite biomarker. In some embodiments, the method includes the step of administering a treatment to the subject. In some embodiments, the method includes, after the step of administering a treatment, making a second determination of the subject at a second time point to obtain at least two variables of the second set. In some embodiments, the method includes calculating at least two variables of the second set to produce a second composite biomarker. In some embodiments, the second composite biomarker can be calculated based on at least two variables of the second set. In some embodiments, the method includes determining whether the treatment is therapeutically effective for treating the subject based on the first and second composite biomarkers. In some embodiments, the method includes comparing the first and second composite biomarkers to obtain a difference between the first and second composite biomarkers. In some embodiments, when the difference between the first and second composite biomarkers exceeds a threshold, the treatment is therapeutically effective for the treated subject. In some embodiments, when the difference between the first and second composite biomarkers does not exceed a threshold, the treatment is not therapeutically effective for the treated subject.

[0089] In some embodiments, a method of treating a subject having an eye disease includes, prior to the step of administering treatment, obtaining, in a first determination of the subject, at least two variables of the first set disclosed herein. In some embodiments, the method includes calculating at least two variables of the first set to generate a first composite biomarker. In some embodiments, the method includes, prior to the step of administering treatment, obtaining, at a second time point, in a second determination of the subject, at least two variables of the second set. In some embodiments, the method includes calculating at least two variables of the second set to generate a second composite biomarker. In some embodiments, the method includes comparing the first and second composite biomarkers to obtain a difference between the first and second composite biomarkers. In some embodiments, the method includes calculating a fifth composite biomarker based on at least two variables of the first and second sets. In some embodiments, the method includes administering treatment to the subject. In some embodiments, the method includes, after the step of administering treatment, obtaining, at a third time point, in a third determination of the subject, at least two variables of the third set. In some embodiments, the method includes, after the step of administering treatment, obtaining, at a fourth time point, in a fourth determination of the subject, at least two variables of the fourth set. In some embodiments, the method includes calculating at least two variables of the third set to generate a third composite biomarker. In some embodiments, the method includes calculating at least two variables of the fourth set to generate a fourth composite biomarker. In some embodiments, the method includes comparing the third and fourth composite biomarkers to obtain a difference between the third and fourth composite biomarkers. In some embodiments, the method includes calculating a sixth composite biomarker based on at least two variables of the third and fourth sets. In some embodiments, the method includes determining whether the treatment is therapeutically effective for the treated subject based on the first, second, third, and fourth composite biomarkers. In some embodiments, the method includes comparing the difference between the first and second composite biomarkers and the difference between the third and fourth composite biomarkers to generate a difference value. In some embodiments, when the difference value exceeds a threshold, the treatment is therapeutically effective for the treated subject. In some embodiments, when the difference value does not exceed the threshold, the treatment is not therapeutically effective for the treated subject. In some embodiments, the method includes determining whether the treatment is therapeutically effective for the treated subject based on the fifth and sixth composite biomarkers. In some embodiments, the method includes comparing the fifth and sixth composite biomarkers to obtain their difference. In some embodiments, when the difference between the fifth and sixth composite biomarkers exceeds a threshold, the treatment is therapeutically effective for the treated subject. In some embodiments, when the difference between the fifth and sixth composite biomarkers does not exceed the threshold, the treatment is not therapeutically effective for the treated subject.

[0090] In some embodiments, the duration between the first and second time points at which the first and second measurements are made, or the duration between the third and fourth time points at which the third and fourth measurements are made, can be any time period suitable for measuring at least two variables disclosed herein. In some embodiments, the duration can be any time period from about 1 day to about 1 year, such as about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, 42 days, 43 days, 44 days, 45 days, 46 days, 47 days, 48 days, 49 days, 50 days, 51 days, 52 days, 53 days, 54 days, 55 days, 56 days, 57 days, 58 days, 59 days, 60 days, 61 days, 62 days, 63 days, 64 days, 65 days, 66 days, 67 days, 68 days, 69 days, 70 days, 71 days, 72 days, 73 days, 74 days, 75 days, 76 days, 77 days, 78 days, 79 days, 80 days, 81 days, 82 days, 83 days, 84 days, 85 days, 86 days, 87 days, 88 days, 89 days, 90 days, 91 days, 92 days, 93 days, 94 days, 95 days, 96 days, 97 days, 98 days, 99 days, 100 days, 101 days, 102 days, 103 days, 104 days, 105 days, 106 days, 107 days, 108 days, 109 days, 110 days, 111 days, 112 days, 113 days, 114 days, 115 days, 116 days, 117 days, 118 days, 119 days, 120 days, 121 days, 122 days, 123 days, 124 days, 125 days, 126 days, 127 days, 128 days, 129 days, 130 days, 131 days, 132 days, 133 days, 134 days, 135 days, 136 days, 137 days, 138 days, 139 days, 140 days, 141 days, 142 days, 143 days, 144 days, 145 days, 146 days, 147 days, 148 days, 149 days, 150 days, 151 days, 152 days, 153 days, 154 days, 155 days, 156 days, 157 days, 158 days, 159 days, 160 days, 161 days, 162 days, 163 days, 164 days, 165 days, 166 days, 167 days, 168 days, 169 days, 170 days, 171 days, 172 days, 173 days, 174 days, 175 days, 176 days, 177 days, 178 days, 179 days, 180 days, 181 days, 182 days, 183 days, 184 days, 185 days, 186 days, 187 days, 188 days, 189 days, 190 days, 191 days, 192 days, 193 days, 194 days, 195 days, 196 days, 197 days, 198 days,199 days, 200 days, 201 days, 202 days, 203 days, 204 days, 205 days, 206 days, 207 days, 208 days, 209 days, 210 days, 211 days, 212 days, 213 days, 214 days, 215 days, 216 days, 217 days, 218 days, 219 days, 220 days, 221 days, 222 days, 223 days, 224 days, 225 days, 226 days, 227 days, 228 days, 229 days, 230 days, 231 days, 232 days, 233 days, 234 days, 235 days, 236 days, 237 days, 238 days, 239 days, 240 days, 241 days, 242 days, 243 days, 244 days, 245 days, 246 days, 247 days, 248 days, 249 days, 250 days, 251 days, 252 days, 253 days, 254 days, 255 days, 256 days, 257 days, 258 days, 259 days, 260 days, 261 days, 262 days, 263 days, 264 days, 265 days, 266 days, 267 days, 268 days, 269 days, 270 days, 271 days, 272 days, 273 days, 274 days, 275 days, 276 days, 277 days, 278 days, 279 days, 280 days, 281 days, 282 days, 283 days, 284 days, 285 days, 286 days, 287 days, 288 days, 289 days, 290 days, 291 days, 292 days, 293 days, 294 days, 295 days, 296 days, 297 days, 298 days, 299 days, 300 days, 301 days, 302 days, 303 days, 304 days, 305 days, 306 days, 307 days, 308 days, 309 days, 310 days, 311 days, 312 days, 313 days, 314 days, 315 days, 316 days, 317 days, 318 days, 319 days, 320 days, 321 days, 322 days, 323 days, 324 days, 325 days, 326 days, 327 days, 328 days, 329 days, 330 days, 331 days, 332 days, 333 days, 334 days, 335 days, 336 days, 337 days, 338 days, 339 days, 340 days, 341 days, 342 days, 343 days, 344 days, 345 days, 346 days, 347 days, 348 days, 349 days, 350 days, 351 days, 352 days, 353 days, 354 days, 355 days, 356 days, 357 days, 358 days, 359 days, 360 days, 361 days, 362 days, 363 days, 364 days or 365 days, or any period therebetween. In some embodiments, the duration can be any period from about 1 week to about 530 weeks, such as about 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks,33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, 81 weeks, 82 weeks, 83 weeks, 84 weeks, 85 weeks, 86 weeks, 87 weeks, 88 weeks, 89 weeks, 90 weeks, 91 weeks, 92 weeks, 93 weeks, 94 weeks, 95 weeks, 96 weeks, 97 weeks, 98 weeks, 99 weeks, 100 weeks, 101 weeks, 102 weeks, 103 weeks, 104 weeks, 105 weeks, 106 weeks, 107 weeks, 108 weeks, 109 weeks, 110 weeks, 111 weeks, 112 weeks, 113 weeks, 114 weeks, 115 weeks, 116 weeks, 117 weeks, 118 weeks, 119 weeks, 120 weeks, 121 weeks, 122 weeks, 123 weeks, 124 weeks, 125 weeks, 126 weeks, 127 weeks, 128 weeks, 129 weeks, 130 weeks, 131 weeks, 132 weeks, 133 weeks, 134 weeks, 135 weeks, 136 weeks, 137 weeks, 138 weeks, 139 weeks, 140 weeks, 141 weeks, 142 weeks, 143 weeks, 144 weeks, 145 weeks, 146 weeks, 147 weeks, 148 weeks, 149 weeks, 150 weeks, 151 weeks, 152 weeks, 153 weeks, 154 weeks, 155 weeks, 156 weeks, 157 weeks, 158 weeks, 159 weeks, 160 weeks, 161 weeks, 162 weeks, 163 weeks, 164 weeks, 165 weeks, 166 weeks, 167 weeks, 168 weeks, 169 weeks, 170 weeks, 171 weeks, 172 weeks, 173 weeks, 174 weeks, 175 weeks, 176 weeks, 177 weeks, 178 weeks, 179 weeks, 180 weeks, 181 weeks, 182 weeks, 183 weeks, 184 weeks, 185 weeks, 186 weeks, 187 weeks, 188 weeks, 189 weeks, 190 weeks, 191 weeks, 192 weeks, 193 weeks, 194 weeks, 195 weeks, 196 weeks, 197 weeks, 198 weeks, 199 weeks, 200 weeks, 201 weeks, 202 weeks, 203 weeks, 204 weeks, 205 weeks, 206 weeks, 207 weeks, 208 weeks, 209 weeks, 210 weeks, 211 weeks, 212 weeks, 213 weeks, 214 weeks, 215 weeks, 216 weeks, 217 weeks, 218 weeks, 219 weeks, 220 weeks, 221 weeks, 222 weeks, 223 weeks, 224 weeks, 225 weeks, 226 weeks, 227 weeks, 228 weeks, 229 weeks, 230 weeks, 231 weeks, 232 weeks, 233 weeks, 234 weeks, 235 weeks, 236 weeks, 237 weeks, 238 weeks, 239 weeks, 240 weeks, 241 weeks, 242 weeks, 243 weeks, 244 weeks, 245 weeksWeek 246, Week 247, Week 248, Week 249, Week 250, Week 251, Week 252, Week 253, Week 254, Week 255, Week 256, Week 257, Week 258, Week 259, Week 260, Week 261, Week 262, Week 263, Week 264, Week 265, Week 266, Week 267, Week 268, Week 269, Week 270, Week 271, Week 272, Week 273, Week 274, Week 275, Week 276, Week 277, Week 278, Week 279, Week 280, Week 281, Week 282, Week 283, Week 284, Week 285, Week 286, Week 287, Week 288, Week 289, Week 290, Week 291, Week 292, Week 293, Week 294, Week 295, Week 296, Week 297, Week 298, Week 299, Week 300, Week 301, Week 302, Week 303, Week 304, Week 305, Week 306, Week 307, Week 308, Week 309, Week 310, Week 311, Week 312, Week 313, Week 314, Week 315, Week 316, Week 317, Week 318, Week 319, Week 320, Week 321, Week 322, Week 323, Week 324, Week 325, Week 326, Week 327, Week 328, Week 329, Week 330, Week 331, Week 332, Week 333, Week 334, Week 335, Week 336, Week 337, Week 338, Week 339, Week 340, Week 341, Week 342, Week 343, Week 344, Week 345, Week 346, Week 347, Week 348, Week 349, Week 350, Week 351, Week 352, Week 353, Week 354, Week 355, Week 356, Week 357, Week 358, Week 359, Week 360, Week 361, Week 362, Week 363, Week 364, Week 365, Week 366, Week 367, Week 368, Week 369, Week 370, Week 371, Week 372, Week 373, Week 374, Week 375, Week 376, Week 377, Week 378, Week 379, Week 380, Week 381, Week 382, Week 383, Week 384, Week 385, Week 386, Week 387, Week 388, Week 389, Week 390, Week 391, Week 392, Week 393, Week 394, Week 395, Week 396, Week 397, Week 398, Week 399, Week 400, Week 401, Week 402, Week 403, Week 404, Week 405, Week 406, Week 407, Week 408, Week 409, Week 410, Week 411, Week 412, Week 413, Week 414, Week 415, Week 416, Week 417, Week 418, Week 419, Week 420, Week 421, Week 422, Week 423, Week 424, Week 425, Week 426, Week 427, Week 428, Week 429, Week 430, Week 431, Week 432, Week 433, Week 434, Week 435, Week 436, Week 437, Week 438, Week 439, Week 440, Week 441, Week 442, Week 443, Week 444, Week 445446 weeks, 447 weeks, 448 weeks, 449 weeks, 450 weeks, 451 weeks, 452 weeks, 453 weeks, 454 weeks, 455 weeks, 456 weeks, 457 weeks, 458 weeks, 459 weeks, 460 weeks, 461 weeks, 462 weeks, 463 weeks, 464 weeks, 465 weeks, 466 weeks, 467 weeks, 468 weeks, 469 weeks, 470 weeks, 471 weeks, 472 weeks, 473 weeks, 474 weeks, 475 weeks, 476 weeks, 477 weeks, 478 weeks, 479 weeks, 480 weeks, 481 weeks, 482 weeks, 483 weeks, 484 weeks, 485 weeks, 486 weeks, 487 weeks, 488 weeks, 489 weeks, 490 weeks, 491 weeks, 492 weeks, 493 weeks, 494 weeks, 495 weeks, 496 weeks, 497 weeks, 498 weeks, 499 weeks, 500 weeks, 501 weeks, 502 weeks, 503 weeks, 504 weeks, 505 weeks, 506 weeks, 507 weeks, 508 weeks, 509 weeks, 510 weeks, 511 weeks, 512 weeks, 513 weeks, 514 weeks, 515 weeks, 516 weeks, 517 weeks, 518 weeks, 519 weeks, 520 weeks, 521 weeks, 522 weeks, 523 weeks, 524 weeks, 525 weeks, 526 weeks, 527 weeks, 528 weeks, 529 weeks or 530 weeks, or any time period therebetween. In some embodiments, the durations disclosed above may apply to any two time points in any of the methods disclosed herein, the method comprising the step of making at least two determinations to obtain at least two sets of at least two variables disclosed herein.,

[0091] In some embodiments, a method of treating a subject having an eye disease comprises the process of selecting a treatment for the subject prior to the step of administering the treatment. In some embodiments, the method comprises the step of making determinations on the subject to obtain at least two variables disclosed herein. In some embodiments, the method comprises the step of calculating at least two variables to produce a composite biomarker. In some embodiments, the method comprises the step of determining whether to select a treatment for the subject based on the composite biomarker. In some embodiments, the method comprises the step of selecting a treatment for the subject when the composite biomarker exceeds a threshold. In some embodiments, the method comprises the step of not selecting a treatment for the subject when the composite biomarker does not exceed a threshold. In some embodiments, the method comprises the step of administering a treatment to the subject when a treatment is selected for the subject. In some embodiments, the method comprises the step of not administering a treatment to the subject when a treatment is not selected for the subject.

[0092] In some embodiments, the process of selecting a treatment for a subject includes the step of performing a first measurement on the subject to obtain at least two variables of a first set. In some embodiments, the process of selecting a treatment for a subject includes the step of performing a second measurement on the subject to obtain at least two variables of a second set. In some embodiments, the process of selecting a treatment for a subject includes calculating at least two variables of the first set and the second set to obtain a first composite biomarker and a second composite biomarker. In some embodiments, the process of selecting a subject includes calculating at least two variables of the first set and the second set to obtain a composite biomarker. In some embodiments, the process of selecting a treatment for a subject includes determining whether to select a treatment for the subject based on the first and second composite biomarkers. In some embodiments, the process of selecting a treatment for a subject includes determining whether to select a treatment for the subject based on a composite biomarker calculated from at least two variables of the first set and the second set. In some embodiments, the process of selecting a treatment for a subject includes comparing the first and second composite biomarkers to obtain their difference. In some embodiments, the process of selecting a treatment for a subject includes selecting a treatment for the subject when the difference between the first and second composite biomarkers exceeds a threshold. In some embodiments, the process of selecting a treatment for a subject includes not selecting a treatment for the subject when the difference between the first and second composite biomarkers does not exceed a threshold. In some embodiments, the method includes selecting a treatment for the subject when a composite biomarker calculated from at least two variables of the first set and the second set exceeds a threshold. In some embodiments, the method includes the step of not selecting a treatment for the subject when a composite biomarker calculated from at least two variables of the first set and the second set does not exceed a threshold. In some embodiments, the threshold for the difference between the first and second composite biomarkers may be the same as or different from the threshold for the composite biomarker calculated from at least two variables of the first set and the second set.

[0093] In some embodiments, the process of selecting a treatment for an eye disease for a subject can be combined with one or more steps of the methods disclosed herein, including but not limited to the process of determining whether a treatment or a dose of a treatment is effective for treating a subject based on one or more composite biomarkers calculated from at least two variables disclosed herein, and the process of administering a therapeutically effective dose of a treatment to a subject based on one or more composite biomarkers calculated from at least two variables measured after the subject has received a certain dose of the treatment.

[0094] In some embodiments, one or more steps of the methods disclosed herein are used to monitor the outcome of treatment for an ocular disease of an object. In some embodiments, the method includes determining whether the dose of treatment administered to the object is therapeutically effective for treating the object by calculating at least two variables of the object disclosed herein to generate one or more composite biomarkers disclosed herein. In some embodiments, the method includes additional steps including, but not limited to, performing one or more assays on the object to obtain at least two variables, and calculating at least two variables of the object to generate one or more composite biomarkers.

[0095] In some embodiments, one or more steps of the methods disclosed herein are used to predict the treatment response of an object to treatment for an ocular disease. In some embodiments, the method includes predicting the treatment response of the object to treatment based on one or more composite biomarkers calculated from at least two variables disclosed herein. In some embodiments, the method includes determining whether the object is likely to have a treatment response to the treatment based on one or more composite biomarkers. In some embodiments, the object is likely to have a treatment response to the treatment when the composite biomarker exceeds a threshold. In some embodiments, the object is likely not to have a treatment response to the treatment when the composite biomarker does not exceed the threshold. In some embodiments, the object is likely to have a treatment response to the treatment when the difference between two or more composite biomarkers disclosed herein exceeds a threshold. In some embodiments, the object is likely not to have a treatment response to the treatment when the difference between two or more composite biomarkers disclosed herein does not exceed the threshold. In some embodiments, the method includes additional steps including, but not limited to, performing one or more assays on the object to obtain at least two variables, and calculating at least two variables of the object to generate one or more composite biomarkers.

[0096] In some embodiments, one or more steps of the methods disclosed herein are used to determine the dose of treatment for an object having an ocular disease based on one or more composite biomarkers calculated from at least two variables disclosed herein. In some embodiments, the dose of treatment can be determined by calculating the composite biomarker or the difference between two or more composite biomarkers. In some embodiments, the dose of treatment is related to the value of the composite biomarker or the difference in values between two or more composite biomarkers. In some embodiments, at least two variables disclosed herein are obtained after administering the treatment to the object. In some embodiments, at least two variables disclosed herein are obtained before administering the treatment to the object.

[0097] In some embodiments, one or more steps of the methods disclosed herein are used to monitor the progression of an ocular disease in a subject based on one or more composite biomarkers calculated from at least two variables disclosed herein. In some embodiments, the status of the ocular disease can be determined by calculating a composite biomarker or the difference between two or more composite biomarkers. In some embodiments, the status of the ocular disease is related to the value of a composite biomarker or the difference in values between two or more composite biomarkers. In some embodiments, at least two of the variables disclosed herein are obtained after administration of a treatment to the subject. In some embodiments, at least two of the variables disclosed herein are obtained before administration of a treatment to the subject.

[0098] In some embodiments, the at least two variables include any number of variables suitable for the methods disclosed herein. In some embodiments, the at least two variables include a number of variables in the range of 2 to 50, such as 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, or any number therebetween. In some embodiments, at least one of the at least two variables disclosed herein is related to one or more of visual acuity (VA), a score on a functional vision questionnaire, and measurements obtained from the imaging techniques disclosed herein. In some embodiments, at least one of the at least two variables is related to changes in ocular structure, changes in ocular pathology, changes in retinal atrophy, changes in macular atrophy, changes in macular degeneration, or a combination thereof. In some instances, the at least two variables include a treatment (e.g., drug type, dose, frequency, method of administration), such as treatment with an RBP4 inhibitor. In some instances, the biomarkers provided herein predict a treatment response to a treatment.

[0099] In some embodiments, the functional vision questionnaire score includes the average score of the functional vision questionnaire. In some embodiments, the imaging technique includes one or more of the imaging techniques disclosed herein. In some embodiments, at least one of the at least two variables is related to one or more selected from changes in visual acuity, changes in the functional vision questionnaire score, and changes in the measurements obtained from the imaging technique. In some embodiments, the change in the functional vision questionnaire score includes a change in the average score of the functional vision questionnaire. In some embodiments, the imaging technique includes a technique for evaluating the eye structure or visual function. In some embodiments, the imaging technique includes a technique for detecting or quantifying changes in the eye structure or visual function. In some embodiments, at least one of the at least two variables is related to changes in visual acuity, changes in the average score of the functional vision questionnaire, and changes in the measurements obtained from the imaging technique for detecting or quantifying changes in optic nerve atrophy or visual function. In some embodiments, the at least two variables are related to changes in visual acuity, changes in the average score of the functional vision questionnaire, and changes in the measurements obtained from the imaging technique for detecting or quantifying changes in optic nerve atrophy or visual function.

[0100] In some embodiments, the at least two variables are related to visual acuity and the average score of the functional vision questionnaire. In some embodiments, the at least two variables are related to changes in visual acuity and changes in the average score of the functional vision questionnaire. In some embodiments, the at least two variables are related to visual acuity and the measurements obtained from the imaging technique for detecting or quantifying changes in optic nerve atrophy or visual function. In some embodiments, the at least two variables are related to changes in visual acuity and changes in the measurements obtained from the imaging technique for detecting or quantifying changes in optic nerve atrophy or visual function. In some embodiments, the at least two variables are related to the average score of the functional vision questionnaire and the measurements obtained from the imaging technique for detecting or quantifying changes in optic nerve atrophy or visual function. In some embodiments, the at least two variables are related to changes in the average score of the functional vision questionnaire and changes in the measurements obtained from the imaging technique for detecting or quantifying changes in optic nerve atrophy or visual function. In some embodiments, both of the at least two variables are related to visual acuity. In some embodiments, both of the at least two variables are related to changes in visual acuity. In some embodiments, both of the at least two variables are related to the average score of the functional vision questionnaire. In some embodiments, both of the at least two variables are related to changes in the average score of the functional vision questionnaire. In some embodiments, both of the at least two variables are related to the measurements obtained from the imaging technique for detecting or quantifying changes in optic nerve atrophy or visual function. In some embodiments, both of the at least two variables are related to changes in the measurements obtained from the imaging technique for detecting or quantifying changes in optic nerve atrophy or visual function.

[0101] In some embodiments, two of at least two variables are related to each other. In some embodiments, the correlation between two of at least two variables can be represented using a correlation coefficient. In some embodiments, a correlation coefficient formula is used to evaluate the strength of the correlation relationship between two of at least two variables. In some embodiments, the correlation coefficient formula can generate a value between -1 and 1. In some embodiments, the value one (1) can represent a strong positive correlation. In some embodiments, the value negative one (-1) can represent a strong negative correlation. In some embodiments, the value zero (0) can represent no relationship. In some embodiments, one of at least two variables is related to visual acuity. In some embodiments, one of at least two variables is related to a change in visual acuity. In some embodiments, one of at least two variables is related to a score on a functional vision questionnaire. In some embodiments, one of at least two variables is related to a change in the score on a functional vision questionnaire. In some embodiments, one of at least two variables is related to an average score on a functional vision questionnaire. In some embodiments, one of at least two variables is related to a change in the average score on a functional vision questionnaire. In some embodiments, one of at least two variables is related to a measurement obtained from an imaging technique used to detect or quantify a change in optic nerve atrophy or visual function. In some embodiments, one of at least two variables is related to a change in a measurement obtained from an imaging technique used to detect or quantify a change in optic nerve atrophy or visual function. In some embodiments, at least two variables include two variables of visual acuity that are related to each other. In some embodiments, at least two variables include two variables of a change in visual acuity that are related to each other. In some embodiments, at least two variables include two variables of an average score on a functional vision questionnaire that are related to each other. In some embodiments, at least two variables include two variables of a change in the average score on a functional vision questionnaire that are related to each other. In some embodiments, at least two variables include two variables of a measurement obtained from an imaging technique used to detect or quantify a change in optic nerve atrophy or visual function that are related to each other. In some embodiments, at least two variables include two variables of a change in a measurement obtained from an imaging technique used to detect or quantify a change in optic nerve atrophy or visual function that are related to each other. In some embodiments, all of at least two variables are related to visual acuity. In some embodiments, all of at least two variables are related to a change in visual acuity. In some embodiments, all of at least two variables are related to an average score on a functional vision questionnaire. In some embodiments, all of at least two variables are related to a change in the average score on a functional vision questionnaire. In some embodiments, all of at least two variables are related to a measurement obtained from an imaging technique used to detect or quantify a change in optic nerve atrophy or visual function.In some embodiments, all of at least two variables are related to a change in a measurement obtained from an imaging technique for detecting or quantifying a change in optic nerve atrophy or visual function. In some embodiments, one of at least two variables is related to two of visual acuity, an average score of a functional vision questionnaire, and a measurement obtained from an imaging technique for detecting or quantifying a change in optic nerve atrophy or visual function. In some embodiments, one of at least two variables is related to two of a change in visual acuity, a change in an average score of a functional vision questionnaire, and a change in a measurement obtained from an imaging technique for detecting or quantifying a change in optic nerve atrophy or visual function. In some embodiments, one of at least two variables is related to each of visual acuity, an average score of a functional vision questionnaire, and a measurement obtained from an imaging technique for detecting or quantifying a change in optic nerve atrophy or visual function. In some embodiments, one of at least two variables is related to each of a change in visual acuity, a change in an average score of a functional vision questionnaire, and a change in a measurement obtained from an imaging technique for detecting or quantifying a change in optic nerve atrophy or visual function. In some embodiments, one of at least two variables is related to the state of an eye disease. In some embodiments, one of at least two variables is related to the progression of an eye disease.

[0102] In some embodiments, at least two variables include values obtained from a log minimum angle of resolution (logMAR) chart, a Snellen chart, a best corrected visual acuity (BCVA) test, an Early Treatment Diabetic Retinopathy Study (ETDRS) letter test, or a combination thereof. In some embodiments, at least two variables include two values obtained from a log minimum angle of resolution (logMAR) chart, a Snellen chart, a best corrected visual acuity (BCVA) test, and an Early Treatment Diabetic Retinopathy Study (ETDRS) letter test.

[0103] In some embodiments, at least two variables include scores of a functional vision questionnaire that includes one or more questions related to one or more of the following groups: the confidence of the subject in moving to different places during the day, the confidence of the subject in moving to different places at night, the ability of the subject to recognize people, the ability of the subject to participate in sports, the ability of the subject to obtain information, the ability of the subject to engage in social interaction, and the ability of the subject to be productive at work. In some embodiments, at least two variables include scores of a functional vision questionnaire that includes one or more questions related to one or more of the following groups: the confidence of the subject in moving to different places during the day or at night, the ability of the subject to recognize people, participate in sports, or obtain information, and the ability of the subject to engage in social interaction or be productive at work. In some embodiments, each of the one or more questions can be scored for the subject. In some embodiments, an average score is obtained from two or more questions of a vision questionnaire test, two or more functional vision questionnaires including one or more questions, or both. In some embodiments, the scoring scale of the vision questionnaire can be any range between two numbers. In some embodiments, the scoring scale can be from 0 to 4. In some embodiments, a score is assigned to the answer "never" of a question according to the scoring scale. In some embodiments, a score is assigned to the answer "almost never" of a question according to the scoring scale. In some embodiments, a score is assigned to the answer "sometimes" of a question according to the scoring scale. In some embodiments, a score is assigned to the answer "almost always" of a question according to the scoring scale. In some embodiments, a score is assigned to the answer "always" of a question according to the scoring scale. In some embodiments, according to the 0-4 scoring scale, scores of 0, 1, 2, 3, and 4 are assigned to the answers "never", "almost never", "sometimes", "almost always", and "always" of a question, respectively. In some embodiments, according to the 0-4 scoring scale, scores of 4, 3, 2, 1, and 0 are assigned to the answers "never", "almost never", "sometimes", "almost always", and "always" of a question, respectively. In some embodiments, scores are assigned to the answers "never", "almost never", "sometimes", "almost always", and "always" of a question according to any range.

[0104] In some embodiments, at least two variables include two or more values obtained from microperimetry (MP), perimetry, or both. In some embodiments, MP includes a spatial map of the retinal sensitivity of a subject. In some embodiments, MP includes the average retinal sensitivity of a subject, its variation, or both. In some embodiments, at least two variables include two or more values obtained from two or more selected from fundus photography, fundus angiography (FA), fundus autofluorescence (FAF), spectral-domain optical coherence tomography (SD-OCT), quantitative autofluorescence (qAF), infrared (IR) imaging, optical coherence tomography-angiography (OCT-A), and wide-field imaging. In some embodiments, at least two variables include at least two values, each value obtained from one of the group consisting of fundus photography, fundus angiography (FA), fundus autofluorescence (FAF), spectral-domain optical coherence tomography (SD-OCT), quantitative autofluorescence (qAF), infrared (IR) imaging, optical coherence tomography-angiography (OCT-A), and wide-field imaging. In some embodiments, at least two variables include two or more values selected from the area of questionable decreased autofluorescence (QDAF), the area of determined decreased autofluorescence (DDAF), the area of decreased autofluorescence (DAF), retinal thickness, ellipsoid zone (EZ) defect width, central subfield retinal thickness (CST), outer subfield retinal thickness (OST), and middle subfield retinal thickness (MST). In some embodiments, at least two variables include two or more values selected from the following: the change in the area of questionable decreased autofluorescence (QDAF), the change in the area of determined decreased autofluorescence (DDAF), the change in the area of decreased autofluorescence (DAF), the change in retinal thickness, the change in ellipsoid zone (EZ) defect width, the change in central subfield retinal thickness (CST), the change in outer subfield retinal thickness (OST), and the change in middle subfield retinal thickness (MST). In some embodiments, at least two variables include DAF and EZ defect width. In some embodiments, at least two variables include the change in DAF and the change in EZ defect width. In some embodiments, the area of decreased autofluorescence (DAF) includes the area of questionable decreased autofluorescence (QDAF) and the area of determined decreased autofluorescence (DDAF). In some embodiments, the change in the area of decreased autofluorescence (DAF) includes the change in the area of questionable decreased autofluorescence (QDAF) and the change in the area of determined decreased autofluorescence (DDAF). In some embodiments, at least two variables include two or more selected from visual acuity, QDAF, DAF, and EZ defect width. In some embodiments, at least two variables include two or more selected from the change in visual acuity, the change in QDAF, the change in DAF, and the change in EZ defect width.In some embodiments, at least two variables include visual acuity, QDAF, DAF, and EZ defect width. In some embodiments, at least two variables include changes in visual acuity, changes in QDAF, changes in DAF, and changes in EZ defect width. In some embodiments, the EZ defect width is related to visual acuity. In some embodiments, the change in EZ defect width is related to the change in visual acuity. In some embodiments, the EZ defect width is related to QDAF. In some embodiments, the change in EZ defect width is related to the change in QDAF. In some embodiments, the EZ defect width is related to DAF. In some embodiments, the change in EZ defect width is related to the change in DAF. In some embodiments, visual acuity is related to QDAF. In some embodiments, QDAF includes a measurement of autofluorescence. In some embodiments, the change in visual acuity is related to the change in QDAF. In some embodiments, the change in QDAF includes a measurement of autofluorescence.

[0105] In some embodiments, the EZ defect width is related to visual acuity, QDAF, and DAF. In some embodiments, the change in EZ defect width is related to the change in visual acuity, the change in QDAF, and the change in DAF. In some embodiments, the EZ defect width is related to visual acuity, QDAF, and DAF, with a correlation coefficient greater than about 0.8 and a p-value less than about 0.05. In some embodiments, the change in EZ defect width is related to the change in visual acuity, the change in QDAF, and the change in DAF, with a correlation coefficient greater than about 0.8 and a p-value less than about 0.05. In some embodiments, visual acuity is related to the EZ defect width and QDAF. In some embodiments, the change in visual acuity is related to the change in EZ defect width and the change in QDAF. In some embodiments, visual acuity is related to the EZ defect width and QDAF, with a p-value less than about 0.05. In some embodiments, the change in visual acuity is related to the change in EZ defect width and the change in QDAF, with a p-value less than about 0.05. In some embodiments, at least two variables are obtained from the right and left eyes of an object. In some embodiments, at least two variables are obtained from at least two objects having different genders, races, ages, or combinations thereof. In some embodiments, at least two variables are obtained from a clinical trial. In some embodiments, at least two variables are obtained from at least two phases of a clinical trial.

[0106] In some embodiments, one or more of the composite biomarkers disclosed herein are used as surrogate biomarkers or endpoints for detecting or predicting improvement in visual function. In some embodiments, the methods disclosed herein are used to detect and / or predict changes in visual function in clinical practice or clinical trials. In some embodiments, the methods disclosed herein are used to detect and / or predict visual improvement in clinical practice or clinical trials. In some embodiments, the methods disclosed herein are used to develop algorithms or software. In some embodiments, the methods disclosed herein can be applied to medical devices, virtual reality goggles, the metaverse, video games, or combinations thereof.

[0107] In some embodiments, the methods disclosed herein can be applied to detect and measure multidimensional visual function. In some embodiments, multidimensional visual function includes spatial awareness, eye-body coordination, or visual motion. In some embodiments, the methods disclosed herein can be applied to detect and measure visual loss or improvement as a multidimensional or multifactorial measurement. In some embodiments, the methods disclosed herein can be applied to develop algorithms, software, or scoring systems for diagnosing eye diseases, detecting visual function improvement or loss, or detecting treatment response during clinical trials.

[0108] In some embodiments, the methods disclosed herein can be applied to develop prediction models related to the variables disclosed herein. In some embodiments, the prediction model can be constructed by a link function based on the magnitude of the variance of each response variable as a function of its predicted value. In some embodiments, the prediction model includes three components. In some embodiments, these three components include a predictor based on a regression function (η i ). In some embodiments, these three components include a model structure that best fits the conditional distribution of the response variable Y i given the values of the explanatory variables in a given model. In some embodiments, these three components include a smooth and invertible link function g(·) that transforms the expectation of the response variable μ i = E(Y i ) into a predictor: g(μ i ) = η i = α + β 1 X i1 + β 2 X i2 + ··· + β k X ik。In some embodiments, a prediction model can be constructed using visual acuity (VA) and EZ width as follows: VA = -0.0896EZ width + 0.0701, Z-score = -2.211, p-value = 0.027, and CI 95% [-0.169, -0.010]. In some embodiments, another univariate or multivariate model can be constructed using visual acuity and EZ width.

[0109] In some embodiments, the methods disclosed herein are used for any eye disease. In some embodiments, the eye disease includes one or more selected from macular lesions, retinopathy, retinal atrophy, macular atrophy, retinal degeneration, macular degeneration, age-related macular degeneration (AMD), RP (retinitis pigmentosa), ABCA4 gene mutation, Stargardt disease (STGD), and STGD1. In some embodiments, the eye disease includes retinal degeneration. In some embodiments, the eye disease includes macular degeneration. In some embodiments, the eye disease includes AMD. In some embodiments, the eye disease includes STGD. In some embodiments, the eye disease includes STGD1.

[0110] In some embodiments, an eye disease can be diagnosed based on one or more factors disclosed herein. In some embodiments, a sample from a subject with an eye disease can have an expression level of retinol binding protein 4 (RBP4) that is at least a threshold. In some embodiments, the threshold for RBP4 can be any value for diagnosing the macular degeneration disease disclosed herein. In some embodiments, the threshold level of RBP4 is at least 25 μg / mL. In some embodiments, the threshold level of RBP4 is at least 30 μg / mL. In some embodiments, the threshold level of RBP4 is at least 35 μg / mL. In some embodiments, the threshold is from about 25 μg / mL to about 100 μg / mL. In some embodiments, the threshold is from about 25 μg / mL to about 30 μg / mL, from about 25 μg / mL to about 35 μg / mL, from about 25 μg / mL to about 40 μg / mL, from about 25 μg / mL to about 50 μg / mL, from about μg / mL to about μg / mL, from about μg / mL to about μg / mL, from about 25 μg / mL to about 80 μg / mL, from about 25 μg / mL to about 90 μg / mL, from about 25 μg / mL to about 100 μg / mL, from about 30 μg / mL to about 35 μg / mL, from about 30 μg / mL to about 40 μg / mL, from about 30 μg / mL to about 50 μg / mL, from about 30 μg / mL to about 60 μg / mL, from about 30 μg / mL to about 70 μg / mL, from about 30 μg / mL to about 80 μg / mL, from about 30 μg / mL to about 90 μg / mL, from about 30 μg / mL to about 100 μg / mL, from about 35 μg / mL to about 40 μg / mL, from about 35 μg / mL to about 50 μg / mL, from about 35 μg / mL to about 60 μg / mL, from about 35 μg / mL to about 70 μg / mL, from about 35 μg / mL to about 80 μg / mL, from about 35 μg / mL to about 90 μg / mL, or from about 35 μg / mL to about 100 μg / mL. In some embodiments, the threshold is about 25 μg / mL, about 30 μg / mL, or about 35 μg / mL. In some embodiments, the threshold is at least about 25 μg / mL, about 30 μg / mL, or about 35 μg / mL. In some embodiments, the threshold level of RBP4 disclosed herein can be applicable to the eye diseases disclosed herein. In some embodiments, the threshold level of RBP4 disclosed herein can be applicable to AMD, STGD, or STGD1. In some embodiments, the threshold is at most about 30 μg / mL, about 35 μg / mL, about 40 μg / mL, about 50 μg / mL, about 60 μg / mL, about 70 μg / mL, about 80 μg / mL, about 90 μg / mL, or about 100 μg / mL. In some embodiments, the level of RBP4 in a subject can be evaluated by measurement.Any assay suitable for measuring RBP4 levels can be employed (such as antibody assays, mass spectrometry-based assays (e.g., LC / MS), liquid chromatography assays (e.g., HPLC, UPLC), etc.). In some embodiments, the assay includes an antibody assay, an electrophoresis assay, an immunoassay, a radioimmunoassay, a chromatography assay, a mass spectrometry assay, a microarray-based detection assay, a polymerase chain reaction assay, a sequencing assay, an immunohistochemistry assay, or any combination thereof. In some embodiments, the assay includes an antibody assay. In some embodiments, the antibody assay includes an enzyme-linked immunosorbent assay (ELISA).

[0111] In some embodiments, the threshold for RBP4 can vary based on various characteristics of the individual. Non-limiting examples of such characteristics include demographic data, gender, age, medical history, previous diagnosis of ocular conditions, body mass index (BMI), height, weight, systemic conditions, including obesity, hyperlipidemia, hyperglycemia, hypertension, diabetes, cardiovascular disease, or non-alcoholic fatty liver disease.

[0112] In some embodiments, if the level of RBP4 is higher than the threshold, an ocular disease is diagnosed. In some embodiments, if the level of RBP4 is higher than the threshold, a risk score is calculated.

[0113] In some embodiments, if the level of RBP4 is higher than the threshold, a therapy is administered. In some embodiments, the therapy is administered to reduce the level of RBP4. In some embodiments, the therapy includes administering a pharmaceutical composition to the individual. In some embodiments, the therapy includes administering a pharmaceutical compound to the individual. In some embodiments, the pharmaceutical composition contains an RBP4 inhibitor. The RBP4 inhibitor can be any RBP4 inhibitor provided herein.

[0114] In some embodiments, a sample from a subject with an eye disease can have an expression level of vitamin A that is at least a threshold level. In some embodiments, the threshold level of vitamin A is at least about 150 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 175 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 200 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 220 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 221 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 222 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 223 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 224 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 225 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 250 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 300 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 350 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 390 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 391 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 392 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 393 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 394 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 395 ng / mL. In some embodiments, the threshold level of vitamin A is from about 150 ng / mL to about 500 ng / mL.In some embodiments, the threshold level of vitamin A is from about 150 ng / mL to about 175 ng / mL, from about 150 ng / mL to about 200 ng / mL, from about 150 ng / mL to about 225 ng / mL, from about 150 ng / mL to about 250 ng / mL, from about 150 ng / mL to about 300 ng / mL, from about 150 ng / mL to about 350 ng / mL, from about 150 ng / mL to about 400 ng / mL, from about 150 ng / mL to about 450 ng / mL, from about 150 ng / mL to about 500 ng / mL, from about 175 ng / mL to about 200 ng / mL, from about 175 ng / mL to about 225 ng / mL, from about 175 ng / mL to about 250 ng / mL, from about 175 ng / mL to about 300 ng / mL, from about 175 ng / mL to about 350 ng / mL, from about 175 ng / mL to about 400 ng / mL, from about 175 ng / mL to about 450 ng / mL, from about 175 ng / mL to about 500 ng / mL, from about 200 ng / mL to about 225 ng / mL, from about 200 ng / mL to about 250 ng / mL, from about 200 ng / mL to about 300 ng / mL, from about 200 ng / mL to about 350 ng / mL, from about 200 ng / mL to about 400 ng / mL, from about 200 ng / mL to about 450 ng / mL, from about 200 ng / mL to about 500 ng / mL, from about 225 ng / mL to about 250 ng / mL, from about 225 ng / mL to about 300 ng / mL, from about 225 ng / mL to about 350 ng / mL, from about 225 ng / mL to about 400 ng / mL, from about 225 ng / mL to about 450 ng / mL, from about 225 ng / mL to about 500 ng / mL, from about 250 ng / mL to about 300 ng / mL, from about 250 ng / mL to about 350 ng / mL, from about 250 ng / mL to about 400 ng / mL, from about 250 ng / mL to about 450 ng / mL, from about 250 ng / mL to about 500 ng / mL, from about 300 ng / mL to about 350 ng / mL, from about 300 ng / mL to about 400 ng / mL, from about 300 ng / mL to about 450 ng / mL, from about 300 ng / mL to about 500 ng / mL, from about 350 ng / mL to about 400 ng / mL, from about 350 ng / mL to about 450 ng / mL, from about 350 ng / mL to about 500 ng / mL, from about 400 ng / mL to about 450 ng / mL, from about 400 ng / mL to about 500 ng / mL, or from about 450 ng / mL to about 500 ng / mL.In some embodiments, the threshold level of vitamin A is about 150 ng / mL, about 175 ng / mL, about 200 ng / mL, about 225 ng / mL, about 250 ng / mL, about 300 ng / mL, about 350 ng / mL, about 400 ng / mL, about 450 ng / mL, or about 500 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 150 ng / mL, about 175 ng / mL, about 200 ng / mL, about 225 ng / mL, about 250 ng / mL, about 300 ng / mL, about 350 ng / mL, about 400 ng / mL, or about 450 ng / mL. In some embodiments, the threshold level of vitamin A is at most about 175 ng / mL, about 200 ng / mL, about 225 ng / mL, about 250 ng / mL, about 300 ng / mL, about 350 ng / mL, about 400 ng / mL, about 450 ng / mL, or about 500 ng / mL. In some embodiments, the threshold levels of vitamin A disclosed herein may be applicable to the eye diseases disclosed herein. In some embodiments, the threshold levels of vitamin A disclosed herein may be applicable to AMD, STGD, or STGD1.

[0115] In some embodiments, the threshold may vary based on various characteristics of the individual. Non-limiting examples of such characteristics include demographics, gender, age, medical history, previous diagnosis of an eye condition, body mass index (BMI), height, weight, systemic conditions, including obesity, hyperlipidemia, hyperglycemia, hypertension, diabetes, cardiovascular disease, or non-alcoholic fatty liver disease.

[0116] In some embodiments, the threshold level of vitamin A depends on the age of the individual. In some embodiments, the threshold level of vitamin A in adolescents (e.g., individuals aged 12 - 17 years) is lower than that in adults (e.g., individuals aged 18 years or older). In some embodiments, the threshold level of vitamin A in adolescents is at least about 150 ng / mL, at least about 175 ng / mL, at least about 200 ng / mL, at least about 210 ng / mL, at least about 220 ng / mL, at least about 221 ng / mL, at least about 222 ng / mL, at least about 223 ng / mL, at least about 224 ng / mL, at least about 225 ng / mL, at least about 230 ng / mL, or at least about 250 ng / mL. In some embodiments, the threshold for adults is at least about 300 ng / mL, at least about 325 ng / mL, at least about 350 ng / mL, at least about 375 ng / mL, at least about 380 ng / mL, at least about 385 ng / mL, at least about 390 ng / mL, at least about 391 ng / mL, at least about 392 ng / mL, at least about 393 ng / mL, at least about 394 ng / mL, at least about 395 ng / mL, or at least about 400 ng / mL.

[0117] In some embodiments, if the level of vitamin A is higher than the threshold, an eye disease is diagnosed. In some embodiments, if the level of vitamin A is higher than the threshold, a risk score is calculated.

[0118] In some embodiments, if the level of vitamin A is higher than the threshold, a treatment is administered. In some embodiments, a therapy is administered to lower the level of vitamin A. In some embodiments, the therapy includes administering a pharmaceutical composition to the individual. In some embodiments, the therapy includes administering a pharmaceutical compound to the individual. In some embodiments, the pharmaceutical composition comprises an RBP4 inhibitor. The RBP4 inhibitor can be any RBP4 inhibitor provided herein.

[0119] In some embodiments, vitamin A levels in an individual are evaluated by an assay. Any assay suitable for measuring vitamin A levels can be employed (e.g., antibody assays, mass spectrometry-based assays (e.g., LC / MS), liquid chromatography assays (e.g., HPLC, UPLC), etc.). In some embodiments, the assay includes an antibody assay, an electrophoresis assay, an immunoassay, a radioimmunoassay, a chromatography assay, a mass spectrometry assay, a microarray-based detection assay, a polymerase chain reaction assay, a sequencing assay, an immunohistochemical assay, or any combination thereof. In some embodiments, the assay includes an antibody assay. In some embodiments, the antibody assay includes an enzyme-linked immunosorbent assay (ELISA). In some embodiments, the assay is a chromatography assay. In some embodiments, the assay includes high performance liquid chromatography (HPLC), ultra-high performance liquid chromatography (UPLC), or liquid chromatography / mass spectrometry (LC-MS).

[0120] In some embodiments, the sample includes a blood sample from a subject. The blood sample can be further processed, such as to remove impurities or to leave serum and / or plasma. In some embodiments, the sample is a plasma or serum sample. In some embodiments, the sample is a plasma sample. In some embodiments, the sample is a serum sample. In some embodiments, the level is measured from plasma or serum derived from the blood sample. In some embodiments, the level is measured from plasma derived from the blood sample. In some embodiments, the level is measured from serum derived from the blood sample.

[0121] In some embodiments, the methods disclosed herein include the steps of assessing whether a subject has an eye disease disclosed herein, or whether the subject is likely to develop an eye disease disclosed herein, such as AMD or STGD. In some embodiments, the method includes determining the presence or absence of one or more genomic variants by assay. In some embodiments, one or more genomic variants include at least 1 of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574. In some embodiments, the method includes calculating a risk score for an eye disease using the presence or absence of one or more genomic variants. In some embodiments, the method includes diagnosing an eye disease using the presence or absence of one or more genomic variants. In some embodiments, the method includes determining the presence or absence of one or more genomic variants by assay. In some embodiments, one or more genomic variants include at least 1 of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574. In some embodiments, one or more genomic variants include at least 2 of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574. In some embodiments, one or more genomic variants include at least 3 of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574. In some embodiments, one or more genomic variants include at least 4 of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574. In some embodiments, one or more genomic variants include at least 5 of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574. In some embodiments, one or more genomic variants include at least 6 of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574.In some embodiments, one or more genomic variants include each of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574. In some embodiments, one or more genomic variants include rs4147863. In some embodiments, one or more genomic variants include rs2275029. In some embodiments, one or more genomic variants include rs1800739. In some embodiments, one or more genomic variants include rs4147857. In some embodiments, one or more genomic variants include rs4147856. In some embodiments, one or more genomic variants include rs1801555. In some embodiments, one or more genomic variants include rs1801574. In some embodiments, one or more genomic variants include at least one of the following: rs3747961, rs6666652, rs1800717, rs763108716, rs185601596, rs17110761, rs61748519, rs1801359, rs145766145, rs76258939, rs200551567, rs754765164, rs201602424, rs564661476, rs4147831, rs6657239, rs2297632, rs1801555, rs1762114, rs55860151, rs1800549, rs3112831, rs4147830, rs2297634, or rs4847281. In some embodiments, one or more genomic variants include at least 5 of the following: rs3747961, rs6666652, rs1800717, rs763108716, rs185601596, rs17110761, rs61748519, rs1801359, rs145766145, rs76258939, rs200551567, rs754765164, rs201602424, rs564661476, rs4147831, rs6657239, rs2297632, rs1801555, rs1762114, rs55860151, rs1800549, rs3112831, rs4147830, rs2297634, or rs4847281. In some embodiments, one or more genomic variants include at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 genomic variants.In some embodiments, one or more genomic variants include at least 1 genomic variant from Table A. In some embodiments, one or more genomic variants include at least 5 genomic variants from Table A. In some embodiments, one or more genomic variants include at least 7 genomic variants from Table A. In some embodiments, one or more genomic variants include at least 10 genomic variants from Table A. One or more genomic variants include at least 15 genomic variants from Table A. In some embodiments, one or more genomic variants include at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 genomic variants from Table A.

[0122] In some embodiments, the treatment for an eye disease disclosed herein includes a pharmaceutical composition comprising an RBP4 inhibitor or a compound that reduces the blood RBP4 concentration in an individual. Further provided herein are methods, wherein the pharmaceutical composition comprises a compound having the structure of formula (I):

[0123]

[0124] Wherein: R A 1 、R A 2 、R A 3 、R A 4 And R A 5 Are each independently H, halogen, CF 3 Or C 1 -C 4 Alkyl, wherein two or more of R A 1 、R A 2 、R A 3 、R A 4 And R A 5 Are not H; R A 6 Is H, OH, or halogen; and A A Has the structure:

[0125]

[0126] Wherein α, β, χ, and δ are each independently absent or present, and when present, each is a bond; X is C or N; Z 1 Is N; Z 2 Is N or NRA 9 , wherein R A 9 is H, C 1 -C 4 alkyl or oxetane; B A is a substituted or unsubstituted 5-, 6- or 7-membered ring structure; or a pharmaceutically acceptable salt thereof.

[0127] The present invention further provides a method, wherein the compound has the structure or a pharmaceutically acceptable salt thereof.

[0128] The present invention further provides a method, wherein the compound has the structure or a pharmaceutically acceptable salt thereof.

[0129] The present invention further provides a method, wherein the pharmaceutical composition comprises a compound having the structure of formula (II):

[0130]

[0131] Wherein: ring A B is optionally further substituted benzene; R B 1 is optionally substituted branched C 3 -C 6 alkyl; X B 1 is O, S, SO, SO 2 or NH; X B 2 is a bond or C 1 -C 3 alkylene; ring B B is azetidine or piperidine; X B 3 is CO or SO 2 ; R B 2 is an optionally substituted hydrocarbon group, an optionally substituted heterocyclic group, an optionally substituted hydroxyl group, an optionally substituted mercapto group, cyano, nitro, acyl or halogen atom; or a pharmaceutically acceptable salt thereof.

[0132] The present invention further provides a method, wherein the compound is 4-(3-(2-tert-butylphenoxy)azetidin-1-yl)-4-oxobutanoic acid, 3-{3-[(2-tert-butyl-4-fluorophenoxy)methyl]azetidin-1-yl}-3-oxopropanoic acid, 2-{[3-(2-tert-butyl-4-chlorophenoxy)azetidin-1-yl]carbonyl}pyridine, 4-[3-(2-tert-butyl-4-chlorophenoxy)azetidin-1-yl]-4-oxobutanoic acid, {3-[(2-tert-butyl-4-chlorophenoxy)methyl]azetidin-1-yl}(oxo)acetic acid, {3-[(2-tert-butylphenoxy)methyl]azetidin-1-yl}(oxo)acetic acid, 3-{3-[(2-tert-butylphenoxy)methyl]azetidin-1-yl}-3-oxopropanoic acid, {4-[(2-tert-butyl-4-chlorophenoxy)methyl]piperidin-1-yl}(oxo)acetic acid, [4-(2-tert-butylphenoxy)piperidin-1-yl](oxo)acetic acid or {4-[(2-tert-butylphenoxy)methyl]piperidin-1-yl}(oxo)acetic acid, or a pharmaceutically acceptable salt thereof.

[0133] The present invention further provides a method, wherein the pharmaceutical composition comprises a compound having the structure of formula (III):

[0134]

[0135] Wherein, ring A C is a benzene ring optionally substituted with 1 to 3 substituents selected from the following: (a) a halogen atom and (b) C 1-6 alkyl; ring B C is a piperazine ring optionally substituted with 1 to 3 substituents selected from the following: (a) a halogen atom, (b) C 1 -C 6 alkyl optionally substituted with 1 to 3 halogen atoms and (c) C 1 -C 6 alkoxy optionally substituted with 1 to 3 halogen atoms; and R C is (1) optionally substituted C 1 -C 10 alkyl, (2) optionally substituted C 6 -C 14 aryl, (3) optionally substituted 5- or 6-membered aromatic heterocyclic group, (4) optionally substituted amino group, (5) optionally substituted carboxyl group or (6) optionally substituted carbamoyl group, or a pharmaceutically acceptable salt thereof.

[0136] The present invention further provides a method, wherein the compound is N-{[4-(2-tert-butylphenyl)piperazin-1-yl]carbonyl}glycine, 3-[4-(2-tert-butylphenyl)piperazin-1-yl]-3-oxopropanoic acid, [4-(2-tert-butyl-4-chlorophenyl)piperazin-1-yl](oxo)acetic acid, 5-{2-[4-(2-tert-butylphenyl)piperazin-1-yl]-2-oxoethyl}imidazolidine-2,4-dione, [(5-{[4-(2-tert-butylphenyl)piperazin-1-yl]carbonyl}isoxazol-3-yl)oxy]acetic acid or a pharmaceutically acceptable salt thereof.

[0137] The present invention further provides a method, wherein the pharmaceutical composition comprises a compound having the structure of formula (IV):

[0138]

[0139] Wherein: Ring A D is a 5-membered non-aromatic heterocycle, which is optionally further substituted by an oxo group; Ring B D is a benzene ring, which is optionally further substituted by 1 to 4 substituents; and X D is O, CH 2 O, OCH 2 , CH 2 , (CH 2 ) 2 , S, CH 2 S, SCH 2 , S(O), CH 2 S(O), S(O)CH 2 , S(O) 2 , CH 2 S(O) 2 or S(O) 2 CH 2 ; or a pharmaceutically acceptable salt thereof.

[0140] The present invention further provides a method, wherein the compound is ((3S)-1-[3,5-bis(trifluoromethyl)phenyl]pyrrolidin-3-yl)oxy)acetic acid, ((1-[4-chloro-3-(trifluoromethyl)phenyl]pyrrolidin-3-yl)thio)acetic acid, 3-((2R,5S)-5-[3,5-bis(trifluoromethyl)phenyl]tetrahydrofuran-2-yl)propanoic acid, or a pharmaceutically acceptable salt thereof.

[0141] The present invention further provides a method, wherein the pharmaceutical composition comprises a compound having the structure of formula (V):

[0142]

[0143] Wherein: Ring A Eis a pyrazole ring, a pyridine ring, an oxazole ring, an imidazole ring or a pyrimidine ring; X E is S, optionally substituted alkylene or O; and R E is a hydrogen atom or C 1 -C 6 alkyl; or a pharmaceutically acceptable salt thereof.

[0144] The present invention further provides methods wherein the compound is ((4-(3,5-bis(trifluoromethyl)phenyl)-1,3-oxazol-2-yl)thio)acetic acid, ethyl ((6-(3,5-bis(trifluoromethyl)phenyl)pyridin-3-yl)thio)acetate, ((6-(3,5-bis(trifluoromethyl)phenyl)pyridin-3-yl)thio)acetic acid, 3-(3-(3,5-bis(trifluoromethyl)phenyl)-1H-pyrazol-1-yl)butyric acid or 3-{3-[3,5-bis(trifluoromethyl)phenyl]-1H-pyrazol-1-yl}propionic acid (also known as STG-001).

[0145] In some embodiments, the treatment comprising the compounds described herein is administered for prophylactic and / or therapeutic treatment. In certain therapeutic applications, the treatment is administered in an amount sufficient to cure or at least partially inhibit at least one symptom of a disease or condition to a patient already suffering from the disease or condition. The effective amount for such use depends on the severity and course of the disease or condition, previous therapy, the health status, body weight and response of the patient to the drug, and the judgment of the treating physician. The therapeutically effective amount is optionally determined by methods including but not limited to dose escalation and / or dose range clinical trials.

[0146] In prophylactic applications, a composition comprising the compounds described herein is administered to a patient susceptible to or at risk of developing a particular disease, disorder or condition. Such amount is defined as a "prophylactically effective amount or dose". In such use, the exact amount also depends on the health status, body weight, etc. of the patient. When used in a patient, the effective amount for such use will depend on the severity and course of the disease, disorder or condition, previous therapy, the health status and response of the patient to the drug, and the judgment of the treating physician. In one aspect, prophylactic treatment includes administering to a mammal (wherein the mammal has previously experienced at least one symptom of the disease being treated and is currently in a state of remission) a pharmaceutical composition comprising the compounds described herein or a pharmaceutically acceptable salt thereof to prevent the recurrence of symptoms of the disease or condition.

[0147] In certain embodiments where the patient's condition does not improve, based on the judgment of the physician, the administration of the compound is a long-term administration, i.e., for an extended period of time, including for the entire duration of the patient's life, in order to improve or otherwise control or limit the symptoms of the patient's disease or condition.

[0148] Oral dosages generally range from about 1.0 mg to about 1000 mg, one to four or more times per day. However, generally speaking, dosages for adult treatment usually fall within the range of 0.01 mg to 5000 mg per day. In one aspect, the dosage for adult treatment is about 1 mg to about 1000 mg per day. In one embodiment, the required dosage is conveniently administered in a single dose or divided doses simultaneously or at appropriate intervals, such as two, three, four or more sub-doses per day.

[0149] In one embodiment, the daily dosage applicable to the compounds or their pharmaceutically acceptable salts described herein is about 0.01 mg / kg to about 50 mg / kg of body weight. In some embodiments, based on many variables related to an individual's treatment regimen, the daily dosage or amount of the active substance in the dosage form is lower or higher than the ranges shown herein. In various embodiments, the daily dosage and unit dosage vary based on many variables, including but not limited to the activity of the compound used, the disease or condition to be treated, the mode of administration, the needs of the individual subject, the severity of the disease or condition being treated, and the judgment of the practicing physician.

[0150] Once the patient's condition improves based on the clinical assessment methods described herein, a maintenance dosage is administered if necessary. Subsequently, in a specific embodiment, the dosage or frequency or both are reduced based on symptoms to a level that maintains the improved disease, disorder or condition. However, in certain embodiments, when any symptoms recur, the patient requires long-term intermittent treatment.

[0151] The toxicity and therapeutic efficacy of such treatment regimens are determined in cell cultures or experimental animals by standard pharmaceutical procedures, including but not limited to the determination of LD50 and ED50. The dose ratio of toxicity to therapeutic effect is the therapeutic index, expressed as the ratio of LD50 to ED50. In certain embodiments, data obtained from cell culture assays and animal studies are used to formulate a therapeutically effective daily dosage range and / or a therapeutically effective unit dosage for mammals (including humans). In some embodiments, the daily dosage of the compounds described herein is within the circulating concentration range that includes the ED50 with minimal toxicity. In certain embodiments, the daily dosage range and / or unit dosage vary within this range, depending on the dosage form employed and the route of administration used.

[0152] In any of the above aspects, there are further embodiments, wherein an effective amount of the compound described herein or a pharmaceutically acceptable salt thereof is: (a) administered systemically to a mammal; and / or (b) administered orally to a mammal; and / or (c) administered intravenously to a mammal; and / or (d) administered by injection to a mammal; and / or (e) administered topically to a mammal; and / or (f) administered non-systemically or locally to a mammal. In some embodiments, an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof is administered orally or parenterally to a subject in need thereof. As used herein, parenteral administration includes intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, intrasternal, intracranial, intramuscular, intrasynovial, intravesical and subcutaneous administration. In some embodiments, an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof is administered orally or intravenously to a subject in need thereof. In some embodiments, an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof is administered orally to a subject in need thereof. In some embodiments, an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof is administered intravenously to a subject in need thereof.

[0153] In any of the above aspects, there are further embodiments, including a single administration of an effective amount of the compound, including further embodiments wherein (i) the compound is administered once daily; or (ii) the compound is administered to the mammal multiple times over the course of a day, such as two, three, four or more times per day. In some embodiments, the RBP4 inhibitory compound described herein is administered daily, every other day, three times a week every other day, every two weeks, every three weeks, every four weeks, every five weeks, every three days, every four days, every five days, every six days, once a week, twice a week (bi-weekly), three times a week, four times a week, five times a week, six times a week, once a month, twice a month, three times a month, once every two months, once every three months, once every four months, once every five months or once every six months. In some embodiments, the RBP4 inhibitory compound described herein or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof is administered daily.

[0154] In any of the above aspects, there are further embodiments that include administering an effective amount of the compound multiple times, including further embodiments where (i) the compound is administered continuously or intermittently: e.g., as a single dose; (ii) the time between multiple administrations is every 3 hours; (iii) the time between multiple administrations is every 6 hours; (iv) the compound is administered to a mammal every 8 hours; (v) the compound is administered to a mammal every 12 hours; or (vi) the compound is administered to a mammal every 24 hours.

[0155] In certain embodiments, where the patient's condition does improve, the dose of the administered drug is temporarily reduced or temporarily discontinued for a period of time (e.g., a "drug holiday"). In a specific embodiment, the length of the drug holiday ranges from 2 days to 1 year, including (by way of example only) 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, or more than 28 days. The dose reduction during the drug holiday (by way of example only) is 10% - 100%, including (by way of example only) 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, and 100%. In a further or alternative embodiment, the method includes a drug holiday where the administration of the compound is temporarily discontinued or the dose of the administered compound is temporarily reduced; at the end of the drug holiday, the administration of the compound is resumed. In one embodiment, the length of the drug holiday varies from 2 days to 7 days. In one embodiment, the length of the drug holiday is 7 days. In one embodiment, the length of the drug holiday is 14 days. In one embodiment, the length of the drug holiday is 28 days.

[0156] Retinol binding protein 4

[0157] In one aspect, the present disclosure provides a method for diagnosing or assessing the risk of developing age-related macular degeneration (AMD) in an individual in need thereof, which includes determining the level of retinol binding protein 4 (RBP4) in a sample from the individual by assay. In some embodiments, the diagnostic or risk assessment step is accompanied by a treatment step, such as administering a therapy to reduce the RBP4 level in the individual, thereby treating or preventing AMD in the individual. In some embodiments, if the RBP4 level in the sample is higher than a threshold (e.g., 25 μg / mL), then a therapy is administered. In some embodiments, if the RBP4 level in the sample is higher than a threshold (e.g., 25 μg / mL), then the individual is diagnosed or assessed as being at risk of developing AMD.

[0158] In another aspect, the present disclosure provides a method for assessing the likelihood of age-related macular degeneration in an individual, which includes determining, by assay, that the level of retinol-binding protein 4 (RBP4) in a sample from the individual is higher than an RBP4 threshold, and assessing the likelihood of developing macular degeneration based on the RBP4 level. In some embodiments, the method further includes determining the age and / or medical history of the individual. Thus, in some embodiments, assessing the likelihood of developing macular degeneration is based on the individual's RBP4 level, age, and medical history.

[0159] In another aspect, the present disclosure provides a method for assessing the severity of age-related macular degeneration in an individual, which includes determining, by assay, that the level of retinol-binding protein 4 (RBP4) in a sample from the individual is higher than an RBP4 threshold, and assessing the severity of macular degeneration based on the RBP4 level. In some embodiments, the method further includes determining the age and / or medical history of the individual. Thus, in some embodiments, assessing the severity of macular degeneration is based on the individual's RBP4 level, age, and medical history.

[0160] In another aspect, the present disclosure provides a method for assessing the diagnosis of age-related macular degeneration in an individual, which includes determining, by assay, that the level of retinol-binding protein 4 (RBP4) in a sample from the individual is higher than an RBP4 threshold, and assessing the diagnosis of macular degeneration based on the RBP4 level. In some embodiments, the method further includes determining the age and / or medical history of the individual. Thus, in some embodiments, assessing the diagnosis of macular degeneration is based on the individual's RBP4 level, age, and medical history.

[0161] In another aspect, the present disclosure provides a method for assessing the dosing regimen of an individual with age-related macular degeneration, which includes determining, by assay, that the level of retinol-binding protein 4 (RBP4) in a sample from the individual is higher than an RBP4 threshold, and assessing the diagnosis of macular degeneration based on the RBP4 level. In some embodiments, the method further includes determining the age and / or medical history of the individual. Thus, in some embodiments, assessing the dosing regimen of an individual with macular degeneration is based on the individual's RBP4 level, age, and medical history.

[0162] In some embodiments, the threshold level of RBP4 is at least 25 μg / mL. In some embodiments, the threshold level of RBP4 is at least 30 μg / mL. In some embodiments, the threshold level of RBP4 is at least 35 μg / mL. In some embodiments, the threshold is from about 25 μg / mL to about 100 μg / mL. In some embodiments, the threshold is from about 25 μg / mL to about 30 μg / mL, from about 25 μg / mL to about 35 μg / mL, from about 25 μg / mL to about 40 μg / mL, from about 25 μg / mL to about 50 μg / mL, from about μg / mL to about μg / mL, from about μg / mL to about μg / mL, from about 25 μg / mL to about 80 μg / mL, from about 25 μg / mL to about 90 μg / mL, from about 25 μg / mL to about 100 μg / mL, from about 30 μg / mL to about 35 μg / mL, from about 30 μg / mL to about 40 μg / mL, from about 30 μg / mL to about 50 μg / mL, from about 30 μg / mL to about 60 μg / mL, from about 30 μg / mL to about 70 μg / mL, from about 30 μg / mL to about 80 μg / mL, from about 30 μg / mL to about 90 μg / mL, from about 30 μg / mL to about 100 μg / mL, from about 35 μg / mL to about 40 μg / mL, from about 35 μg / mL to about 50 μg / mL, from about 35 μg / mL to about 60 μg / mL, from about 35 μg / mL to about 70 μg / mL, from about 35 μg / mL to about 80 μg / mL, from about 35 μg / mL to about 90 μg / mL, or from about 35 μg / mL to about 100 μg / mL. In some embodiments, the threshold is about 25 μg / mL, about 30 μg / mL, or about 35 μg / mL. In some embodiments, the threshold is at least about 25 μg / mL, about 30 μg / mL, or about 35 μg / mL. In some embodiments, the threshold is at most about 30 μg / mL, about 35 μg / mL, about 40 μg / mL, about 50 μg / mL, about 60 μg / mL, about 70 μg / mL, about 80 μg / mL, about 90 μg / mL, or about 100 μg / mL.

[0163] In some embodiments, the threshold can vary based on various characteristics of the individual. Non-limiting examples of such characteristics include demographics, sex, age, medical history, previous diagnosis of an ocular condition, body mass index (BMI), height, weight, systemic conditions, including obesity, hyperlipidemia, hyperglycemia, hypertension, diabetes, cardiovascular disease, or non-alcoholic fatty liver disease.

[0164] In some embodiments, if the RBP4 level is above the threshold, a diagnosis of AMD is made. In some embodiments, if the RBP4 level is above the threshold, a risk score is calculated.

[0165] In some embodiments, if the RBP4 level is above a threshold, a therapy is administered. In some embodiments, the therapy is administered to reduce the RBP4 level. In some embodiments, the therapy includes administering a pharmaceutical composition to an individual. In some embodiments, the therapy includes administering a pharmaceutical compound to an individual. In some embodiments, the pharmaceutical composition comprises an RBP4 inhibitor. The RBP4 inhibitor can be any RBP4 inhibitor provided herein.

[0166] In some embodiments, if the RBP4 level is below a threshold, additional steps may be taken or recommended. Such below-threshold readings may still indicate some risk of the individual developing AMD in the future, and thus future monitoring may be recommended or additional tests may be required to confirm or complete the diagnosis of AMD.

[0167] In addition, in some embodiments, the method includes assessing the likelihood of developing AMD based on the RBP4 level. In some embodiments, assessing the likelihood of developing AMD includes generating a risk score. In some embodiments, if the risk score is above a threshold, an AMD treatment (e.g., treatment with an RBP4 inhibitor) is administered. In some embodiments, the risk score reflects the likelihood of the individual developing AMD at a future time point based on the measurements provided herein.

[0168] In some embodiments, the method includes providing a recommendation to re-evaluate the individual's AMD after a period of time (e.g., when a risk score for developing AMD has been calculated or when the level of RBP4 is below a threshold but there are other signs indicating a possible development of AMD). In some embodiments, if the level or risk score is below a threshold, the method includes providing a recommendation to re-evaluate the individual's AMD after a period of time. The period of time can be any suitable period of time. In some embodiments, the period of time is from about 1 month to about 24 months. In some embodiments, the period of time is from about 1 month to about 3 months, from about 1 month to about 6 months, from about 1 month to about 9 months, from about 1 month to about 12 months, from about 1 month to about 18 months, from about 1 month to about 24 months, from about 3 months to about 6 months, from about 3 months to about 9 months, from about 3 months to about 12 months, from about 3 months to about 18 months, from about 3 months to about 24 months, from about 6 months to about 9 months, from about 6 months to about 12 months, from about 6 months to about 18 months, from about 6 months to about 24 months, from about 9 months to about 12 months, from about 9 months to about 18 months, from about 9 months to about 24 months, from about 12 months to about 18 months, from about 12 months to about 24 months, or from about 18 months to about 24 months. In some embodiments, the period of time is about 1 month, about 3 months, about 6 months, about 9 months, about 12 months, about 18 months, or about 24 months. In some embodiments, the period of time is at least about 1 month, about 3 months, about 6 months, about 9 months, about 12 months, or about 18 months. In some embodiments, the period of time is at most about 3 months, about 6 months, about 9 months, about 12 months, about 18 months, or about 24 months.

[0169] The recommendation for re-evaluation includes performing additional tests after a period of time, including but not limited to re-evaluating the individual's RBP4 level or performing any other AMD test or evaluation provided herein, such as a physician follow-up for an eye physical examination.

[0170] In some embodiments, the RBP4 level in an individual is evaluated by an assay. Any assay suitable for measuring the RBP4 level can be employed (e.g., antibody assay, mass spectrometry-based assay (e.g., LC / MS), liquid chromatography assay (e.g., HPLC, UPLC), etc.). In some embodiments, the assay includes an antibody assay, an electrophoresis assay, an immunoassay, a radioimmunoassay, a chromatography assay, a mass spectrometry assay, a microarray-based detection assay, a polymerase chain reaction assay, a sequencing assay, an immunohistochemistry assay, or any combination thereof. In some embodiments, the assay includes an antibody assay. In some embodiments, the antibody assay includes an enzyme-linked immunosorbent assay (ELISA).

[0171] In some embodiments, the sample comprises a blood sample from an individual. The blood sample can be further processed, such as to remove impurities or to leave serum and / or plasma. In some embodiments, the sample is a plasma or serum sample. In some embodiments, the sample is a plasma sample. In some embodiments, the sample is a serum sample. In some embodiments, the level is measured from plasma or serum derived from the blood sample. In some embodiments, the level is measured from plasma derived from the blood sample. In some embodiments, the level is measured from serum derived from the blood sample.

[0172] In some embodiments, in addition to the RBP4 level, the determination of AMD diagnosis, the assessment of the risk of developing AMD, and / or the decision to administer a therapy are made based on other considerations. In some embodiments, the determination is based on the age and / or medical history of the individual. In some embodiments, the determination is at least partially based on the age of the individual. In some embodiments, the determination is at least partially based on the medical history of the individual. In some embodiments, the medical history of the individual can include information such as a previously diagnosed eye condition, body mass index (BMI), height, weight, systemic conditions (including obesity, hyperlipidemia, hyperglycemia, hypertension, diabetes, cardiovascular disease, or non-alcoholic fatty liver disease), etc. In some embodiments, the determination is at least partially based on the vitamin A level in the individual (e.g., in a blood sample of the individual).

[0173] In addition, the methods provided herein can further include any additional tests or diagnoses for assessing the presence of AMD in the individuals provided herein, including the measurement of the presence or absence of any genomic variants provided herein or any other measurements useful for diagnosing AMD. In some embodiments, the method further includes performing at least one of color fundus photography, fundus autofluorescence, spectral-domain optical coherence tomography, or microperimetry.

[0174] In some embodiments, the method further includes classifying the progression of age-related macular degeneration. The classification can be based on the level of RBP4 (measured as provided herein) or in combination with other methods provided herein (such as by using the Age-Related Eye Disease Study (AREDS) classification).

[0175] Vitamin A Levels Predictive of AMD

[0176] In one aspect, the present disclosure provides a method for diagnosing or assessing the risk of developing age-related macular degeneration (AMD) in an individual in need thereof, which includes determining the vitamin A level in a sample from the individual by measurement. In some embodiments, the diagnosing or risk assessment step is accompanied by a treatment step, such as administering a therapy to reduce the vitamin A or RBP4 level in the individual, thereby treating or preventing AMD in the individual. In some embodiments, if the vitamin A level in the sample is higher than a threshold (e.g., 150 ng / mL), a therapy is administered. In some embodiments, if the vitamin A level in the sample is higher than a threshold (e.g., 150 ng / mL), the individual is diagnosed or assessed as being at risk of developing AMD.

[0177] In another aspect, the present disclosure provides a method for assessing the likelihood of age-related macular degeneration in an individual, which includes determining by measurement that the vitamin A level in a sample from the individual is higher than a threshold of vitamin A, and assessing the likelihood of developing macular degeneration based on the vitamin A level. In some embodiments, the method further includes determining the age and / or medical history of the individual. Thus, in some embodiments, the assessment of the likelihood of developing macular degeneration is based on the individual's vitamin A level, age, and medical history.

[0178] In another aspect, the present disclosure provides a method for assessing the severity of age-related macular degeneration in an individual, which includes determining by measurement that the vitamin A level in a sample from the individual is higher than a threshold of vitamin A, and assessing the severity of macular degeneration based on the vitamin A level. In some embodiments, the method further includes determining the age and / or medical history of the individual. Thus, in some embodiments, the assessment of the severity of macular degeneration is based on the individual's vitamin A level, age, and medical history.

[0179] In another aspect, the present disclosure provides a method for assessing the diagnosis of age-related macular degeneration in an individual, which includes determining by measurement that the vitamin A level in a sample from the individual is higher than a threshold of vitamin A, and assessing the diagnosis of macular degeneration based on the vitamin A level. In some embodiments, the method further includes determining the age and / or medical history of the individual. Thus, in some embodiments, the assessment of the diagnosis of macular degeneration is based on the individual's vitamin A level, age, and medical history.

[0180] In another aspect, the present disclosure provides a method for assessing a dosing regimen in an individual with age-related macular degeneration, which includes determining by measurement that the vitamin A level in a sample from the individual is higher than a threshold of vitamin A, and assessing the diagnosis of macular degeneration based on the vitamin A level. In some embodiments, the method further includes determining the age and / or medical history of the individual. Thus, in some embodiments, the assessment of the dosing regimen for an individual with macular degeneration is based on the individual's vitamin A level, age, and medical history.

[0181] In some embodiments, the threshold level of vitamin A is at least about 150 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 175 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 200 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 220 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 221 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 222 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 223 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 224 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 225 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 250 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 300 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 350 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 390 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 391 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 392 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 393 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 394 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 395 ng / mL. In some embodiments, the threshold level of vitamin A is from about 150 ng / mL to about 500 ng / mL.In some embodiments, the threshold level of vitamin A is from about 150 ng / mL to about 175 ng / mL, from about 150 ng / mL to about 200 ng / mL, from about 150 ng / mL to about 225 ng / mL, from about 150 ng / mL to about 250 ng / mL, from about 150 ng / mL to about 300 ng / mL, from about 150 ng / mL to about 350 ng / mL, from about 150 ng / mL to about 400 ng / mL, from about 150 ng / mL to about 450 ng / mL, from about 150 ng / mL to about 500 ng / mL, from about 175 ng / mL to about 200 ng / mL, from about 175 ng / mL to about 225 ng / mL, from about 175 ng / mL to about 250 ng / mL, from about 175 ng / mL to about 300 ng / mL, from about 175 ng / mL to about 350 ng / mL, from about 175 ng / mL to about 400 ng / mL, from about 175 ng / mL to about 450 ng / mL, from about 175 ng / mL to about 500 ng / mL, from about 200 ng / mL to about 225 ng / mL, from about 200 ng / mL to about 250 ng / mL, from about 200 ng / mL to about 300 ng / mL, from about 200 ng / mL to about 350 ng / mL, from about 200 ng / mL to about 400 ng / mL, from about 200 ng / mL to about 450 ng / mL, from about 200 ng / mL to about 500 ng / mL, from about 225 ng / mL to about 250 ng / mL, from about 225 ng / mL to about 300 ng / mL, from about 225 ng / mL to about 350 ng / mL, from about 225 ng / mL to about 400 ng / mL, from about 225 ng / mL to about 450 ng / mL, from about 225 ng / mL to about 500 ng / mL, from about 250 ng / mL to about 300 ng / mL, from about 250 ng / mL to about 350 ng / mL, from about 250 ng / mL to about 400 ng / mL, from about 250 ng / mL to about 450 ng / mL, from about 250 ng / mL to about 500 ng / mL, from about 300 ng / mL to about 350 ng / mL, from about 300 ng / mL to about 400 ng / mL, from about 300 ng / mL to about 450 ng / mL, from about 300 ng / mL to about 500 ng / mL, from about 350 ng / mL to about 400 ng / mL, from about 350 ng / mL to about 450 ng / mL, from about 350 ng / mL to about 500 ng / mL, from about 400 ng / mL to about 450 ng / mL, from about 400 ng / mL to about 500 ng / mL, or from about 450 ng / mL to about 500 ng / mL.In some embodiments, the threshold level of vitamin A is about 150 ng / mL, about 175 ng / mL, about 200 ng / mL, about 225 ng / mL, about 250 ng / mL, about 300 ng / mL, about 350 ng / mL, about 400 ng / mL, about 450 ng / mL, or about 500 ng / mL. In some embodiments, the threshold level of vitamin A is at least about 150 ng / mL, about 175 ng / mL, about 200 ng / mL, about 225 ng / mL, about 250 ng / mL, about 300 ng / mL, about 350 ng / mL, about 400 ng / mL, or about 450 ng / mL. In some embodiments, the threshold level of vitamin A is at most about 175 ng / mL, about 200 ng / mL, about 225 ng / mL, about 250 ng / mL, about 300 ng / mL, about 350 ng / mL, about 400 ng / mL, about 450 ng / mL, or about 500 ng / mL.

[0182] In some embodiments, the threshold may vary based on various characteristics of the individual. Non-limiting examples of such characteristics include demographics, gender, age, medical history, previous diagnosis of eye conditions, body mass index (BMI), height, weight, systemic conditions, including obesity, hyperlipidemia, hyperglycemia, hypertension, diabetes, cardiovascular disease, or non-alcoholic fatty liver disease.

[0183] In some embodiments, the threshold level of vitamin A depends on the age of the individual. In some embodiments, the vitamin A threshold level in adolescents (e.g., individuals aged 12 - 17 years) is lower than that in adults (e.g., individuals aged 18 years or older). In some embodiments, the vitamin A threshold level in adolescents is at least about 150 ng / mL, at least about 175 ng / mL, at least about 200 ng / mL, at least about 210 ng / mL, at least about 220 ng / mL, at least about 221 ng / mL, at least about 222 ng / mL, at least about 223 ng / mL, at least about 224 ng / mL, at least about 225 ng / mL, at least about 230 ng / mL, or at least about 250 ng / mL. In some embodiments, the threshold in adults is at least about 300 ng / mL, at least about 325 ng / mL, at least about 350 ng / mL, at least about 375 ng / mL, at least about 380 ng / mL, at least about 385 ng / mL, at least about 390 ng / mL, at least about 391 ng / mL, at least about 392 ng / mL, at least about 393 ng / mL, at least about 394 ng / mL, at least about 395 ng / mL, or at least about 400 ng / mL.

[0184] In some embodiments, if the vitamin A level is above a threshold, AMD is diagnosed. In some embodiments, if the vitamin A level is above a threshold, a risk score is calculated.

[0185] In some embodiments, if the vitamin A level is above a threshold, a therapy is administered. In some embodiments, the therapy is administered to reduce the vitamin A level. In some embodiments, the therapy includes administering a pharmaceutical composition to an individual. In some embodiments, the therapy includes administering a pharmaceutical compound to an individual. In some embodiments, the pharmaceutical composition comprises an RBP4 inhibitor. The RBP4 inhibitor can be any RBP4 inhibitor provided herein.

[0186] In some embodiments, if the vitamin A level is below a threshold, additional steps may be taken or recommended. Such below-threshold readings may still indicate some risk of the individual developing AMD in the future, and thus future monitoring may be recommended or additional tests may be required to confirm or complete the diagnosis of AMD.

[0187] Furthermore, in some embodiments, the method includes assessing the likelihood of developing AMD based on the vitamin A level. In some embodiments, assessing the likelihood of developing AMD includes generating a risk score. In some embodiments, if the risk score is above a threshold, an AMD treatment (e.g., treatment with an RBP4 inhibitor) is administered. In some embodiments, the risk score reflects the likelihood of the individual developing AMD at a future time point based on the measurements provided herein.

[0188] In some embodiments, the method includes providing a recommendation to re-evaluate an individual's AMD after a period of time (e.g., when a risk score for developing AMD has been calculated or when the vitamin A level is below a threshold but there are other signs indicating possible development of AMD). In some embodiments, if the level or risk score is below a threshold, the method includes providing a recommendation to re-evaluate an individual's AMD after a period of time. The period of time can be any suitable period of time. In some embodiments, the period of time is from about 1 month to about 24 months. In some embodiments, the period of time is from about 1 month to about 3 months, from about 1 month to about 6 months, from about 1 month to about 9 months, from about 1 month to about 12 months, from about 1 month to about 18 months, from about 1 month to about 24 months, from about 3 months to about 6 months, from about 3 months to about 9 months, from about 3 months to about 12 months, from about 3 months to about 18 months, from about 3 months to about 24 months, from about 6 months to about 9 months, from about 6 months to about 12 months, from about 6 months to about 18 months, from about 6 months to about 24 months, from about 9 months to about 12 months, from about 9 months to about 18 months, from about 9 months to about 24 months, from about 12 months to about 18 months, from about 12 months to about 24 months, or from about 18 months to about 24 months. In some embodiments, the period of time is about 1 month, about 3 months, about 6 months, about 9 months, about 12 months, about 18 months, or about 24 months. In some embodiments, the period of time is at least about 1 month, about 3 months, about 6 months, about 9 months, about 12 months, or about 18 months. In some embodiments, the period of time is at most about 3 months, about 6 months, about 9 months, about 12 months, about 18 months, or about 24 months.

[0189] The recommendation for re-evaluation includes performing additional tests after a period of time, including but not limited to re-evaluating the individual's vitamin A level or performing any other AMD test or evaluation provided herein, such as a physician follow-up for an eye physical examination.

[0190] In some embodiments, an individual's vitamin A level is evaluated by an assay. Any assay suitable for measuring vitamin A level can be employed (e.g., antibody assay, mass spectrometry-based assay (e.g., LC / MS), liquid chromatography assay (e.g., HPLC, UPLC), etc.). In some embodiments, the assay includes an antibody assay, an electrophoresis assay, an immunoassay, a radioimmunoassay, a chromatography assay, a mass spectrometry assay, a microarray-based detection assay, a polymerase chain reaction assay, a sequencing assay, an immunohistochemistry assay, or any combination thereof. In some embodiments, the assay includes an antibody assay. In some embodiments, the antibody assay includes an enzyme-linked immunosorbent assay (ELISA). In some embodiments, the assay is a chromatography assay. In some embodiments, the assay includes high performance liquid chromatography (HPLC), ultra high performance liquid chromatography (UPLC), or liquid chromatography / mass spectrometry (LC-MS).

[0191] In some embodiments, the sample comprises a blood sample from an individual. The blood sample can be further processed, such as to remove impurities or to leave serum and / or plasma. In some embodiments, the sample is a plasma or serum sample. In some embodiments, the sample is a plasma sample. In some embodiments, the sample is a serum sample. In some embodiments, the level is measured from plasma or serum derived from the blood sample. In some embodiments, the level is measured from plasma derived from the blood sample. In some embodiments, the level is measured from serum derived from the blood sample.

[0192] In some embodiments, in addition to the vitamin A level, the determination of AMD diagnosis, the risk assessment of developing AMD, and / or the decision to administer a therapy are made based on other considerations. In some embodiments, the determination is based on the age and / or medical history of the individual. In some embodiments, the determination is at least partially based on the age of the individual. In some embodiments, the determination is at least partially based on the medical history of the individual. In some embodiments, the medical history of the individual can include information such as a previously diagnosed eye condition, body mass index (BMI), height, weight, systemic conditions (including obesity, hyperlipidemia, hyperglycemia, hypertension, diabetes, cardiovascular disease, or non-alcoholic fatty liver disease). In some embodiments, the determination is at least partially based on the RBP4 level in the individual (e.g., in a blood sample of the individual).

[0193] In addition, the methods provided herein can further include any additional tests or diagnoses for assessing the presence of AMD in the individuals provided herein, measurements for determining the presence or absence of any genomic variants provided herein, or any other measurements useful for diagnosing AMD. In some embodiments, the method further includes performing at least one of color fundus photography, fundus autofluorescence, spectral-domain optical coherence tomography, or microperimetry.

[0194] In some embodiments, the method further includes classifying the progression of age-related macular degeneration. The classification can be based on the level of vitamin A (measured as provided herein) or in combination with other methods provided herein (such as by using the Age-Related Eye Disease Study (AREDS) classification).

[0195] Genomic Variants Predictive of AMD

[0196] The present disclosure also provides genomic variants indicative of an individual's propensity to develop AMD. In some embodiments, the genomic variants include single nucleotide polymorphisms (SNPs) in the gene encoding ATP-binding cassette subfamily A member 4 (ABCA4) in an individual. ABCA4 is a protein encoded by the ABCA4 gene in humans and other eukaryotes. The ABCA4 protein is expressed almost exclusively in the retina and is associated with Stargardt and other ocular diseases, including but not limited to macular dystrophy, cone-rod dystrophy, retinitis pigmentosa, and age-related macular degeneration. Reduced ABCA4 activity is associated with the excessive accumulation of toxic retinoids and lipofuscin. In certain cases, such mutations are detected by sequencing the DNA or RNA of the subject. In some embodiments, the genomic variants provided herein can predict the likelihood that a subject will develop AMD, either alone or in combination with other factors. The genomic variants herein include SNPs that are missense SNPs, intronic SNPs, synonymous SNPs, or any other type of SNP. An individual includes one or more of the SNPs provided herein, located on the same allele or different alleles. Examples of such genomic variants and SNPs can be found in Table A below.

[0197] Table A. Genomic Variants

[0198] Variant number Variant Mutation Protein change Type 1 rs3747961 c.*372＝ NA 3’UTR variant 2 rs6666652 c.6764G>T p.S2255I Missense 3 rs1800717 NA NA Intron 4 rs763108716 c.6579C>T p.F2193= Synonymous 5 rs185601596 NA NA Intron 6 rs17110761 NA NA Intron 7 rs61748519 c.6255C>T p.L2085= Synonymous 8 rs1801359 c.6249C>T p.12083＝ Synonymous 9 rs145766145 NA NA Intron 10 rs76258939 c.3626T>C p.M1209T Missense 11 rs200551567 c.2841C>T p.D947= Synonymous 12 rs754765164 NA NA Intron splice acceptor variant 13 rs201602424 c.1614C>T p.A538= Synonymous 14 rs564661476 NA NA Intron insertion 15 rs4147831 c.1269C>T p.H423= Synonymous 16 rs6657239 c.635G>A p.R212H Missense 17 rs2297632 NA NA Intron 18 rs1801555 c.6285T>C p.D2095= Synonymous 19 rs1762114 c.6069T>C p.I2023= Synonymous 20 rs4147863％ NA NA Intron 21 rs2275029％ c.5844A>G p.P1948= Synonymous 22 rs1800739％ NA NA Intron 23 rs4147857％ c.5814A>G p.L1938= Synonymous 24 rs4147856 NA NA Intron 25 rs1801574 c.5682G>C p.L1894= Synonymous 26 rs55860151 NA NA Intron 27 rs1800549 c.4283C>T p.T1428M Missense 28 rs3112831 c.1268A>C p.H423R Synonymous 29 rs4147830 NA NA Intron 30 rs2297634 NA NA Intron 31 rs4847281 c.141A>G p.P47= Synonymous

[0199] In one aspect, the present disclosure provides a method of assessing whether an individual has AMD, is likely to develop AMD, and, in certain cases, treating or preventing the AMD. In some embodiments, the method includes determining the presence or absence of one or more genomic variants by assay. In some embodiments, the one or more genomic variants include at least one of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574. In some embodiments, the method includes using the presence or absence of the one or more genomic variants to calculate a risk score for age-related macular degeneration. In some embodiments, the method includes using the presence or absence of the one or more genomic variants to diagnose age-related macular degeneration. In some embodiments, the method further includes administering a therapy to treat the individual's age-related macular degeneration. In some embodiments, the method further includes administering a therapy to prevent the individual's age-related macular degeneration.

[0200] In some embodiments, the method includes determining the presence or absence of one or more genomic variants by assay. In some embodiments, the one or more genomic variants include at least 1 of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574. In some embodiments, the one or more genomic variants include at least 2 of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574. In some embodiments, the one or more genomic variants include at least 3 of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574. In some embodiments, the one or more genomic variants include at least 4 of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574. In some embodiments, the one or more genomic variants include at least 5 of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574. In some embodiments, the one or more genomic variants include at least 6 of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574. In some embodiments, the one or more genomic variants include each of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574.

[0201] In some embodiments, the one or more genomic variants include rs4147863. In some embodiments, the one or more genomic variants include rs2275029. In some embodiments, the one or more genomic variants include rs1800739. In some embodiments, the one or more genomic variants include rs4147857. In some embodiments, the one or more genomic variants include rs4147856. In some embodiments, the one or more genomic variants include rs1801555. In some embodiments, the one or more genomic variants include rs1801574.

[0202] In some embodiments, one or more genomic variants include at least one of the following: rs3747961, rs6666652, rs1800717, rs763108716, rs185601596, rs17110761, rs61748519, rs1801359, rs145766145, rs76258939, rs200551567, rs754765164, rs201602424, rs564661476, rs4147831, rs6657239, rs2297632, rs1801555, rs1762114, rs55860151, rs1800549, rs3112831, rs4147830, rs2297634, or rs4847281. In some embodiments, one or more genomic variants comprise at least 5 of the following: rs3747961, rs6666652, rs1800717, rs763108716, rs185601596, rs17110761, rs61748519, rs1801359, rs145766145, rs76258939, rs200551567, rs754765164, rs201602424, rs564661476, rs4147831, rs6657239, rs2297632, rs1801555, rs1762114, rs55860151, rs1800549, rs3112831, rs4147830, rs2297634, or rs4847281. In some embodiments, the one or more genomic variants include at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 genomic variants.

[0203] In some embodiments, one or more genomic variants include at least 1 genomic variant from Table A. In some embodiments, one or more genomic variants include at least 5 genomic variants from Table A. In some embodiments, one or more genomic variants include at least 7 genomic variants from Table A. In some embodiments, one or more genomic variants include at least 10 genomic variants from Table A. One or more genomic variants include at least 15 genomic variants from Table A. In some embodiments, one or more genomic variants include at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 genomic variants from Table A.

[0204] In some embodiments, detecting the presence or absence of one or more genomic variants further comprises determining the allele frequency of one or more genomic variants in an individual. In some embodiments, one or more genomic variants are present on a single copy of an individual gene. In some embodiments, one or more genomic variants are present on multiple copies of an individual gene.

[0205] In some embodiments, determining the presence or absence of one or more genomic variants comprises assaying genetic material from an individual. In some cases, the genetic material is obtained from blood, serum, plasma, sweat, hair, tears, urine, and other techniques known to those of skill in the art. In some embodiments, the genetic material is obtained from the blood, serum, or plasma of an individual.

[0206] In some embodiments, methods of detecting the presence, absence, or level of genomic variants in a sample obtained from an individual involve detecting nucleic acid sequences. In some cases, the nucleic acid sequence comprises deoxyribonucleic acid (DNA), such as in the case of detecting complementary DNA (cDNA) of an mRNA transcript. In some cases, the nucleic acid sequence comprises a denatured DNA molecule or a fragment thereof. In some cases, the nucleic acid sequence comprises DNA selected from the group consisting of: genomic DNA, amplified DNA, circular DNA, circulating DNA, cell-free DNA, or exosomal DNA. In some cases, the DNA is single-stranded DNA (ssDNA), double-stranded DNA, denatured double-stranded DNA, synthetic DNA, and combinations thereof. Circular DNA can be cleaved or fragmented. In some cases, the nucleic acid sequence comprises ribonucleic acid (RNA). In some cases, the nucleic acid sequence comprises fragmented RNA. In some cases, the nucleic acid sequence comprises partially degraded RNA.

[0207] In some embodiments, disclosed herein are methods of detecting genomic variants by nucleic acid-based assays of samples obtained from a subject. In some cases, the nucleic acid-based assays include quantitative polymerase chain reaction (qPCR), gel electrophoresis (including, for example, Northern or Southern blotting), immunochemistry, in situ hybridization (such as fluorescence in situ hybridization (FISH)), cytochemistry, microarrays, or sequencing. In some embodiments, the sequencing techniques include next-generation sequencing techniques. In some embodiments, the method involves hybridization assays, such as fluorescence qPCR (e.g., TaqMan TM, SYBR green, SYBRgreen I, SYBR green II, SYBR gold, ethidium bromide, methylene blue, Pyronin Y, DAPI, acridine orange, BlueView, or phycoerythrin), which involves a nucleic acid amplification reaction with a specific primer pair and hybridization of an amplified nucleic acid probe containing a detectable moiety or molecule specific for the target nucleic acid sequence. In some cases, the number of amplification cycles used to detect the target nucleic acid in a qPCR assay is from about 5 to about 30 cycles. In some cases, the number of amplification cycles used to detect the target nucleic acid is at least about 5 cycles. In some cases, the number of amplification cycles used to detect the target nucleic acid is at most about 30 cycles. In some cases, the number of amplification cycles used to detect the target nucleic acid is from about 5 to about 10, about 5 to about 15, about 5 to about 20, about 5 to about 25, about 5 to about 30, about 10 to about 15, about 10 to about 20, about 10 to about 25, about 10 to about 30, about 15 to about 20, about 15 to about 25, about 15 to about 30, about 20 to about 25, about 20 to about 30, or about 25 to about 30 cycles. For TaqMan TM method, the probe can be a hydrolyzable probe that contains a fluorophore and a quencher and is hydrolyzed by DNA polymerase when hybridized to the target nucleic acid. In some cases, the presence of the target nucleic acid is determined when the number of amplification cycles reaching the threshold is less than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, or 20 cycles. In some cases, hybridization can occur at a standard hybridization temperature, such as from about 35°C to about 65°C in a standard PCR buffer.

[0208] Another exemplary nucleic acid-based assay involves the use of nucleic acid probes that are bound or otherwise immobilized on beads, microtiter plates, or other substrates, where the nucleic acid probes are configured to hybridize to a target nucleic acid sequence. In some cases, the nucleic acid probes are specific for one or more of the gene products (PRS) described herein. In some cases, nucleic acid probes specific for a biomarker include nucleic acid probe sequences that are sufficiently complementary to the polynucleotide sequence of the biomarker. In some cases, the biomarker includes a transcribed polynucleotide sequence (e.g., RNA, cDNA). In some embodiments, the nucleic acid probe can be, for example, a full-length cDNA or a portion thereof, such as an oligonucleotide having a length of at least about 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides and sufficient to specifically hybridize to the target nucleic acid sequence under standard hybridization conditions. In some embodiments, the target nucleic acid sequence is immobilized on a solid surface and contacted with the probe, for example, by running the separated target nucleic acid sequence on an agarose gel and transferring the target nucleic acid sequence from the gel to a membrane (such as nitrocellulose). In some embodiments, the probe is immobilized on a solid surface, such as in an Affymetrix gene chip array, and the probe is contacted with the target nucleic acid sequence.

[0209] In some embodiments, the term "probe" with respect to nucleic acids refers to any nucleic acid molecule capable of selectively binding to a specific target nucleic acid sequence. In some cases, probes are specifically designed to be labeled, for example, with a radioactive label, a fluorescent label, an enzyme, a chemiluminescent tag, a colorimetric tag, or other labels or tags known in the art. In some cases, the fluorescent label includes a fluorophore. In some cases, the fluorophore is an aromatic or heteroaromatic compound. In some cases, the fluorophore is pyrene, anthracene, naphthalene, acridine, stilbene, benzoxazole, indole, benzindole, oxazole, thiazole, benzothiazole, cyanine, carbocyanine, salicylate, anthranilate, xanthene dyes, coumarin. Exemplary xanthene dyes include, for example, fluorescein and rhodamine dyes. Fluorescein and rhodamine dyes include, but are not limited to, 6-carboxyfluorescein (FAM), 2’7’-dimethoxy-4’5’-dichloro-6-carboxyfluorescein (JOE), tetrachlorofluorescein (TET), 6-carboxyrhodamine (R6G), N,N,N;N’-tetramethyl-6-carboxyrhodamine (TAMRA), 6-carboxy-X-rhodamine (ROX). Suitable fluorescent probes also include naphthylamine dyes having an amino group at the α or β position. For example, naphthylamino compounds include 1-dimethylaminonaphthyl-5-sulfonate, 1-anilinonaphthyl-8-sulfonate, and 2-p-toluidinyl-6-naphthalenesulfonate, 5-(2’-aminoethyl)aminonaphthalene-1-sulfonic acid (EDANS). Exemplary coumarins include, for example, 3-phenyl-7-isothiocyanatocoumarin; acridines such as 9-isothiocyanatoacridine and acridine orange; N-(p-(2-benzoxazolyl)phenyl)maleimide; cyanines such as indodicarbocyanine 3 (Cy3), indodicarbocyanine 5 (Cy5), indodicarbocyanine 5.5 (Cy5.5), 3-(-carboxy-pentyl)-3’-ethyl-5,5’-dimethyloxacarbocyanine (CyA); 1H,5H,11H,15H-xantheno[2,3,4-ij:5,6,7-I’j’]diquinoxalin-18-ium, 9-[2(or 4)-[[[6-[2,5-dioxo-1-pyrrolidinyl)oxy]-6-oxohexyl]amino]sulfonyl]-4(or 2)-sulfophenyl]-2,3,6,7,12,13,16,17-octahydro-inner salt (TR or Texas Red); or BODIPYTM dyes. In certain cases, the probe contains FAM as a dye label.

[0210] In some embodiments, detecting one or more genomic variants comprises sequencing genetic material obtained from a sample of an object. Sequencing can be performed with any suitable sequencing technology, including but not limited to single molecule real-time (SMRT) sequencing, Polony sequencing, ligation sequencing, reversible terminator sequencing, proton detection sequencing, ion semiconductor sequencing, nanopore sequencing, electronic sequencing, pyrosequencing, Maxam-Gilbert sequencing, chain termination (e.g., Sanger) sequencing, +S sequencing, or sequencing by synthesis. Sequencing methods also include next-generation sequencing, such as modern sequencing technologies, such as Illumina sequencing (e.g., Solexa), Roche 454 sequencing, Ion torrent sequencing, and SOLiD sequencing. In some cases, next-generation sequencing involves high-throughput sequencing methods. Other sequencing methods available to those skilled in the art can also be employed.

[0211] In some embodiments, if a genomic variant provided herein is identified in an individual, a therapy is administered. In some embodiments, the therapy is administered to reduce the level of RBP4 in the individual. In some embodiments, the therapy comprises administering a pharmaceutical composition to the individual. In some embodiments, the therapy comprises administering a pharmaceutical compound to the individual. In some embodiments, the pharmaceutical composition comprises an RBP4 inhibitor. The RBP4 inhibitor can be any RBP4 inhibitor provided herein.

[0212] Furthermore, the methods provided herein can also include any additional tests or diagnostics provided herein for assessing the presence of AMD in an individual, including measurement of RBP4 levels and comparison to a threshold provided herein, measurement of vitamin A levels and comparison to a threshold provided herein, or any other measurement that can be used to diagnose AMD. In some embodiments, the method further comprises performing at least one of color fundus photography, fundus autofluorescence, spectral domain-optical coherence tomography, or microperimetry.

[0213] In some embodiments, the method further comprises determining the age and medical history of the individual. Thus, in some embodiments, the likelihood of developing macular degeneration is evaluated based on the presence of one or more genomic variants, age, and medical history of the individual.

[0214] In some embodiments, in addition to the presence or absence of one or more genomic variants, the diagnosis of AMD, the risk assessment of developing AMD, or the treatment decision is determined based on other considerations. In some embodiments, the determination is based on the age and / or medical history of the individual. In some embodiments, the determination is at least partially based on the age of the individual. In some embodiments, the determination is at least partially based on the medical history of the individual. In some embodiments, the medical history of the individual may include information such as previously diagnosed eye conditions, body mass index (BMI), height, weight, systemic conditions (including obesity, hyperlipidemia, hyperglycemia, hypertension, diabetes, cardiovascular disease, or non-alcoholic fatty liver disease), and the like.

[0215] In some embodiments, the identification of one or more genomic variants provided herein in an individual does not necessarily require immediate therapeutic intervention, but may prompt a treating physician to recommend increased monitoring for the development of AMD or related symptoms indicative of the development of AMD. In some embodiments, the method includes providing a recommendation to re-evaluate the AMD of the individual after a period of time. In some embodiments, if the level or risk score is below a threshold, the method includes providing a recommendation to re-evaluate the AMD of the individual after a period of time. The period of time can be any suitable period of time. In some embodiments, the period of time is from about 1 month to about 24 months. In some embodiments, the period of time is from about 1 month to about 3 months, from about 1 month to about 6 months, from about 1 month to about 9 months, from about 1 month to about 12 months, from about 1 month to about 18 months, from about 1 month to about 24 months, from about 3 months to about 6 months, from about 3 months to about 9 months, from about 3 months to about 12 months, from about 3 months to about 18 months, from about 3 months to about 24 months, from about 6 months to about 9 months, from about 6 months to about 12 months, from about 6 months to about 18 months, from about 6 months to about 24 months, from about 9 months to about 12 months, from about 9 months to about 18 months, from about 9 months to about 24 months, from about 12 months to about 18 months, from about 12 months to about 24 months, or from about 18 months to about 24 months. In some embodiments, the period of time is about 1 month, about 3 months, about 6 months, about 9 months, about 12 months, about 18 months, or about 24 months. In some embodiments, the period of time is at least about 1 month, about 3 months, about 6 months, about 9 months, about 12 months, or about 18 months. In some embodiments, the period of time is at most about 3 months, about 6 months, about 9 months, about 12 months, about 18 months, or about 24 months. The period of time can also be a longer time, such as at least about 2 years, at least about 3 years, at least about 5 years, at least about 10 years, or longer. In some embodiments, regular monitoring of the development of AMD in the individual may be recommended.

[0216] Suggestions for re-evaluation include performing additional tests after a period of time, including but not limited to assessing RBP4 levels in an individual or performing any of the other AMD tests or assessments provided herein, such as physician follow-up for an eye physical examination.

[0217] In addition, the methods provided herein can also include any additional tests or diagnostics provided herein for assessing the presence of AMD in an individual, including measurement of RBP4 levels and comparison to the thresholds provided herein, measurement of vitamin A levels and comparison to the thresholds provided herein, or any other measurements that can be used to diagnose AMD. In some embodiments, the method also includes performing at least one of color fundus photography, fundus autofluorescence, spectral domain-optical coherence tomography, or microperimetry.

[0218] In some embodiments, the method also includes classifying the progression of age-related macular degeneration. The classification can be based on the level of RBP4 (measured as provided herein) or in combination with other methods provided herein (such as by using the classification of the Age-Related Eye Disease Study (AREDS)).

[0219] Combination of RBP4 level / vitamin A level and genomic variants for predicting AMD

[0220] It is also possible to simultaneously probe the biomarker levels provided herein (e.g., blood concentration of RBP4 and / or vitamin A) and genomic variants (e.g., ABCA4 variants) to provide a diagnosis, risk score, or treatment recommendation for an individual's AMD. In some embodiments, the diagnostic or risk score generated by this combination method is more reliable than using any one method alone. In some embodiments, this allows for a certain diagnosis or risk assessment of AMD without the need for a more invasive physical examination of the eye, since the diagnostic or risk score can be calculated from a single sample (e.g., a blood sample) from the individual.

[0221] One aspect of the present invention is a method for assessing the likelihood of age-related macular degeneration in an individual in need thereof, comprising: obtaining a blood sample from the individual; extracting a protein fraction from the blood sample; extracting a nucleic acid fraction from the blood sample; determining, by a first assay, the level of retinol-binding protein 4 (RBP4) and / or vitamin A from the protein fraction; determining, by a second assay, the allele frequencies of one or more genomic variants of the nucleic acid from the nucleic acid fraction; and assessing the likelihood of age-related macular degeneration based on the RBP4 level and / or vitamin A level and the allele frequencies of one or more genomic variants. In some embodiments, the one or more genomic variants include at least one of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574.

[0222] Another aspect of the present invention is a method for treating age-related macular degeneration in an individual in need thereof, comprising: a) providing the level of retinol-binding protein 4 (RBP4) and / or the level of vitamin A, wherein the level is determined by an assay of the protein fraction of a sample from the individual; b) providing the allele frequencies of one or more genomic variants, wherein the allele frequencies are determined by an assay of the nucleic acid fraction of a sample from the individual; and c) administering a therapy based on an assessment of the level of RBP4 and / or the level of vitamin A and the allele frequencies of one or more genomic variants. In some embodiments, the one or more genomic variants include at least one of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574.

[0223] In some embodiments, the level of RBP4 is compared to an RBP4 threshold provided elsewhere herein. Any RBP4 threshold provided herein is used in combination with the detection of genomic variants in a diagnostic / assessment protocol. Additionally, any method or assay for determining the level of RBP4 in a sample provided herein is equally applicable to diagnosing AMD, assessing the risk of developing AMD, or determining a method of treating AMD based on a combination of genomic variants and RBP4 level.

[0224] In some embodiments, the methods provided herein are used to detect the presence or absence of one or more genomic variants. Any combination or number of genomic variants provided herein can be used in combination with the information provided by the RBP4 level for the diagnosis, risk assessment, or treatment of AMD provided herein.

[0225] In embodiments for measuring RBP4 levels and genomic variants, an assay measures the RBP4 level and a second assay determines the presence or absence (or allele frequency) of one or more genomic variants in a sample derived from an individual. In some embodiments, the assay and the second assay are performed on the same sample. In some embodiments, the assay and the second assay are performed on different samples derived from the individual. The samples need not be collected from the individual at the same time, and the samples need not be of the same type. In some embodiments, the assay and the second assay are performed on the same sample derived from the individual. In some embodiments, the assay and the second assay are performed on two independent samples derived from the individual. In some embodiments, the two independent samples are samples of the same type (e.g., blood samples). In some embodiments, the two independent samples are samples of different types (e.g., a blood sample and a urine sample). In some embodiments, the two independent samples are two blood samples. The two independent samples need not be collected from the individual at the same time. In some embodiments, the two independent samples are collected from the individual at the same time. In some embodiments, the two independent samples are collected at different times. In some embodiments, the two independent samples are collected at intervals of at most about 1 day, at most about 2 days, at most about 3 days, at most about 1 week, at most about 2 weeks, at most about 1 month, at most about 2 months, at most about 3 months, or at most about 6 months.

[0226] As the methods provided herein for assessing the presence or absence of AMD based on measuring RBP4 levels or genomic variants can further include additional tests or diagnostics to determine the presence of AMD in an individual, so can methods that utilize the presence or absence of RBP4 levels and genomic variants. In some embodiments, the determination of diagnosis, risk assessment, or administration of therapy is also based on additional criteria, such as the age and medical history of the subject. In some embodiments, the determination is based on the age and / or medical history of the individual. In some embodiments, the determination is at least partially based on the age of the individual. In some embodiments, the determination is at least partially based on the medical history of the individual. In some embodiments, the medical history of the individual can include information such as a previously diagnosed eye condition, body mass index (BMI), height, weight, systemic conditions (including obesity, hyperlipidemia, hyperglycemia, hypertension, diabetes, cardiovascular disease, or non-alcoholic fatty liver disease), and the like.

[0227] In addition, the methods provided herein can further include any additional tests or diagnostics provided herein for assessing the presence of AMD in an individual. In some embodiments, the method further includes performing at least one of color fundus photography, fundus autofluorescence, spectral domain-optical coherence tomography, or microperimetry.

[0228] In some embodiments, the method further includes classifying the progression of age-related macular degeneration. The classification can be based on the level of RBP4 (measured as provided herein), or in combination with other methods provided herein (such as by using the Age-Related Eye Disease Study (AREDS) classification).

[0229] In some embodiments, the risk score reflects the probability that an individual will develop AMD at a future time point based on the measurements provided herein.

[0230] In some embodiments, the identification of one or more genomic variants provided herein and the measurement of the level of RBP4 in an individual do not require immediate therapeutic intervention, but can prompt a treating physician to recommend increased monitoring for the development of AMD or related symptoms indicative of the development of AMD. In some embodiments, the method includes providing a recommendation to re-evaluate the individual's AMD after a period of time. In some embodiments, if the level or risk score is below a threshold, the method includes providing a recommendation to re-evaluate the individual's AMD after a period of time. The period of time can be any suitable period of time. In some embodiments, the period of time is from about 1 month to about 24 months. In some embodiments, the period of time is from about 1 month to about 3 months, from about 1 month to about 6 months, from about 1 month to about 9 months, from about 1 month to about 12 months, from about 1 month to about 18 months, from about 1 month to about 24 months, from about 3 months to about 6 months, from about 3 months to about 9 months, from about 3 months to about 12 months, from about 3 months to about 18 months, from about 3 months to about 24 months, from about 6 months to about 9 months, from about 6 months to about 12 months, from about 6 months to about 18 months, from about 6 months to about 24 months, from about 9 months to about 12 months, from about 9 months to about 18 months, from about 9 months to about 24 months, from about 12 months to about 18 months, from about 12 months to about 24 months, or from about 18 months to about 24 months. In some embodiments, the period of time is about 1 month, about 3 months, about 6 months, about 9 months, about 12 months, about 18 months, or about 24 months. In some embodiments, the period of time is at least about 1 month, about 3 months, about 6 months, about 9 months, about 12 months, or about 18 months. In some embodiments, the period of time is at most about 3 months, about 6 months, about 9 months, about 12 months, about 18 months, or about 24 months. The period of time can also be a longer time, such as at least about 2 years, at least about 3 years, at least about 5 years, at least about 10 years, or longer. In some embodiments, regular monitoring of the development of AMD in the individual can be recommended.

[0231] The recommendation for re-evaluation includes performing additional tests after a period of time, including but not limited to assessing the level of RBP4 in the individual or performing any other AMD test or evaluation provided herein, such as a physician follow-up for an eye physical examination.

[0232] In some embodiments, if a genomic variant provided herein is identified in an individual, a therapy is administered. In some embodiments, the therapy is administered to reduce the level of RBP4 in the individual. In some embodiments, the therapy includes administering a pharmaceutical composition to the individual. In some embodiments, the therapy includes administering a pharmaceutical compound to the individual. In some embodiments, the pharmaceutical composition comprises an RBP4 inhibitor. The RBP4 inhibitor can be any RBP4 inhibitor provided herein.

[0233] Additional Considerations for Predicting or Diagnosing AMD

[0234] In some embodiments, determining a diagnosis of AMD, assessing the risk of developing AMD, or deciding whether to treat is based on additional tests that can optionally be performed in addition to measuring the RBP4 level of the subject and / or determining the presence or absence of a genomic variant provided herein. Accordingly, the methods provided herein can also provide additional tests for use in conjunction with the biomarkers indicative of AMD provided herein.

[0235] The methods provided herein can also include additional tests for assessing the presence of AMD or the risk of developing AMD in an individual. A variety of such additional tests and diagnostics are known in the art, including color fundus photography, fundus autofluorescence, spectral domain-optical coherence tomography, and microperimetry.

[0236] In some embodiments, the methods provided herein also include analyzing at least one of color fundus photography, fundus autofluorescence, spectral domain-optical coherence tomography, or microperimetry. In some embodiments, the method also includes analyzing color fundus photography. In some embodiments, the method also includes analyzing fundus autofluorescence. In some embodiments, the method also includes spectral domain-optical coherence tomography. In some embodiments, the method also includes microperimetry. In some embodiments, the diagnosis of AMD, the assessment of the risk of developing AMD, or the decision to administer therapy for AMD is at least partially based on one of these assessments.

[0237] In addition, the methods provided herein can also include determining the age or medical history of the individual. In some embodiments, the determination is at least partially based on the age of the individual. In some embodiments, the determination is at least partially based on the medical history of the individual. In some embodiments, the medical history of the individual can include information such as a previously diagnosed eye condition, body mass index (BMI), height, weight, systemic conditions (including obesity, hyperlipidemia, hyperglycemia, hypertension, diabetes, cardiovascular disease, or non-alcoholic fatty liver disease), and the like.

[0238] In some embodiments, the methods herein include classifying the progression of AMD. In some embodiments, the classification is based on the Age-Related Eye Disease Study Classification [Age-Related Eye Disease Study Research G. The Age-Related Eye Disease Study (AREDS): design implications. AREDS report no. 1. Control Clin Trials. 1999;20(6):573-600) (AREDS)]. The AREDS classification includes Category 1 (defined as few (1-15), small (<63 μm), or no drusen and no pigment changes); Category 2 (early AMD, characterized by the presence of several small, few medium-sized (63-124 μm) drusen and / or pigment changes in one or both eyes); Category 3 (intermediate AMD, characterized by the presence of numerous (20 soft or 65 hard without any soft), medium-sized drusen, one large (>125 μm) drusen, and / or geographic atrophy not involving the macula in one or both eyes); Category 4 (advanced unilateral AMD, consisting of either the late dry form with geographic atrophy involving the macula in one eye or the exudative form with choroidal neovascularization in one eye).

[0239] RBP4 inhibitory compounds and compounds that reduce RBP4 in the blood

[0240] In some embodiments, the present disclosure provides RBP4 inhibitory compounds, compounds that reduce the level of RBP4 in the blood, and pharmaceutical compositions comprising the compounds. In some embodiments, the compound is an RBP4 inhibitor. In some embodiments, the compound reduces the level of RBP4 in the blood. The compounds and compositions herein can be used to inhibit RPB4 and treat various diseases, including age-related macular degeneration and STGD. In some embodiments, after diagnosing an individual with AMD or STGD or assessing the risk of an individual developing AMD or STGD using the methods provided herein, a therapy comprising an RBP4 inhibitory compound (such as those provided herein) is administered to the individual. Examples of RBP4 inhibitor compounds can be found in U.S. Publication No. US2010 / 0292206; PCT Publication No. WO2010119992; U.S. Publication No. US2011 / 0251187, U.S. Publication No. US2018 / 0237404; U.S. Patent No. 10,273,243; U.S. Patent No. 8,980,924; U.S. Patent No. 9,637,450; U.S. Patent No. 9,944,644; U.S. Patent No. 9,938,291; U.S. Patent No. 10,072,016; and PCT Publication No. WO2018232154, which are all incorporated herein by reference.

[0241] Some embodiments provided herein describe an RBP4 inhibitor or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof for treating age-related macular degeneration, having the structure of formula (I):

[0242]

[0243] Wherein:

[0244] R A 1 、R A 2 、R A 3 、R A 4 and R A 5 are each independently halogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclic group, optionally substituted heterocycloalkyl, -COR A 7 、-CON(R A 7 ) 2 、optionally substituted (C 0 -C 4 alkylene)-CN, optionally substituted (C 0 -C 4 alkylene)-OR A 7 、optionally substituted (C 0 -C 4 alkylene)-N(R A 7 ) 2 、optionally substituted (C 0 -C 4 alkylene)N(R A 8 )-COR A 7 、optionally substituted (C 0 -C 4 alkylene)-SO 2 N(R A 7 ) 2 、optionally substituted (C 0 -C 4 alkylene)-SO 2 R A 7 、optionally substituted (C 0 -C 4 alkylene)N(RA 8 )-SO 2 N(R A 7 ) 2 or optionally substituted (C 0 -C 4 alkylene)N(R A 8 )-SO 2 R A 7 ;

[0245] Each R A 7 is independently selected from H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted carbocyclic group, optionally substituted carbocyclic alkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclic group, optionally substituted heterocyclic alkyl, optionally substituted heteroaryl or optionally substituted heteroaralkyl; or two R A 11 groups together with the nitrogen to which they are attached join to form an optionally substituted N-heterocyclic group;

[0246] Each R A 8 is independently selected from H or optionally substituted alkyl;

[0247] R A 6 is -H, -OH, optionally substituted alkyl or halogen;

[0248] p is 0, 1, 2, 3, 4 or 5;

[0249] A A has the structure:

[0250]

[0251] wherein:

[0252] α, β, χ and δ each independently are absent or present, and when present each is a bond;

[0253] X is C;

[0254] Z 1 is S, O or N;

[0255] Z 2 is S, O, N or NR A 9 ;

[0256] R A 9 is H, optionally substituted alkyl or oxetane; and

[0257] B A is a substituted or unsubstituted fused 5-, 6- or 7-membered ring structure; or

[0258] a pharmaceutically acceptable salt thereof.

[0259] In certain embodiments, the compound of formula (I) has the structure

[0260]

[0261] wherein

[0262] R A 1 、R A 2 、R A 3 、R A 4 and R A 5 are each independently H, halogen, CF 3 or C 1 -C 4 alkyl, wherein two or more of R A 1 、R A 2 、R A 3 、R A 4 and R A 5 are not H;

[0263] R A 6 is H, OH or halogen; and

[0264] A A has the structure:

[0265]

[0266] wherein

[0267] α, β, χ and δ each independently are absent or present and, when present, each is a bond;

[0268] X is C or N;

[0269] Z 1 is N;

[0270] Z 2 is N or NR A 9 ,

[0271] wherein RA 9 is H, C 1 -C 4 alkyl or oxetane;

[0272] B A is a substituted or unsubstituted 5-, 6- or 7-membered ring structure; or

[0273] its pharmaceutically acceptable salts.

[0274] In some embodiments, the compound of formula (I) is 1-(3-(4-(3,4-difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)ethan-1-one. In certain embodiments, the compound of formula (I) is 1-(3-(4-(3,4-difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)ethan-1-one; 1-(3-(4-(3,4-difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazol-5(1H)-yl)ethan-1-one; (4-(3-fluoro-2,5-bis(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(2-chloro-3-fluorophenyl)piperidin-1-yl)(5-(2-methoxyethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(2-chloro-3-fluorophenyl)piperidin-1-yl)(5-(3,3,3-trifluoropropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(2-chloro-3-fluorophenyl)piperidin-1-yl)(5-(2,2,2-trifluoroethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(2-chloro-3-fluorophenyl)piperidin-1-yl)(5-(oxetan-3-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(4-fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(4-fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; (4-(2-chloro-3-fluorophenyl)piperidin-1-yl)(5-(cyclopropylmethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(2-chloro-3-fluorophenyl)piperidin-1-yl)(5-ethyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(3,4-difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(6-ethyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; (4-(4-fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(methylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone;1-(3-(4-(2-Fluoro-6-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)ethan-1-one; (4-(3-Fluoro-2,5-bis(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; (4-(2-Chloro-3-fluorophenyl)piperidin-1-yl)(6-(cyclopropylmethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; (4-(3,5-Bis(trifluoromethyl)phenyl)piperidin-1-yl)(5-(methylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(2-Chloro-3-fluorophenyl)piperidin-1-yl)(5-(methylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(2-Chloro-3-fluorophenyl)piperidin-1-yl)(6-(oxetan-3-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; 3-(4-(2-Chloro-3-fluorophenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carbonitrile (4-(5-Fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(methylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; 3-(4-(3,5-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-N-methyl-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxamide (6-(cyclopropylmethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(3,4-difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 3-(4-(3,5-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylic acid methyl ester; (4-(3,5-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(3,3,3-trifluoropropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-neopentyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(2-Chloro-5-fluorophenyl)piperidin-1-yl)(5-(methylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone;(4-(3,5-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(6-(2,2,2-trifluoroethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; 3-(4-(3,5-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carbonitrile; (4-(3,5-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(6-(oxetan-3-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; 3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-N-methyl-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxamide; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(6-neopentyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(6-(2,2,2-trifluoroethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; 3-(4-(2-Chloro-3-fluorophenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylic acid methyl ester; 1-(3-(4-(3,5-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)ethan-1-one; (6-(Cyclopropylmethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(3,5-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(6-(2-methoxyethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; 1-(3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)-3-methylbutan-1-one; 1-(3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)propan-1-one; 1-(3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)-2-methylpropan-1-one;3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carbonitrile; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(6-(3,3,3-trifluoropropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(2-methoxyethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(3,3,3-trifluoropropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; 3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-N-methyl-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxamide; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(6-(oxetan-3-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; 3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylic acid methyl ester; 2-(3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)acetic acid; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(oxetan-3-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (5-(Cyclopropylmethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-ethyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; 3-(4-(3-Fluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-N-methyl-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxamide; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(6-methyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone;Methyl 3-(4-(3,5-difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate; (4-(3,5-difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-ethyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(3,5-difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(2-methoxyethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; 3-(4-(3,4-difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carbonitrile; 3-(4-(3,4-difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate; (4-(3-fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(6-(3,3,3-trifluoropropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; (6-ethyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(3-fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (4-(3-fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(oxetan-3-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(3-fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(2-methoxyethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(3-fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(3,3,3-trifluoropropyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(2-chloro-5-fluorophenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; Imidazo[1,2-a]pyridin-2-yl(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (4-(5-fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; 1-(3-(4-(3,4-difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one;(5-Ethyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(3-fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 3-(4-(3-fluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-N-methyl-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxamide; (4-(3-fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(methylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(3,5-difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(3,5-difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; (4-(2-chloro-3-fluorophenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(2-chloro-3-fluorophenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; (4-(3,4-difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(3,4-difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; (4-(3,5-bis(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(3,5-bis(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; (4-(3-fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(3-fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; 1-(3-(4-(2-chloro-3-fluorophenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)ethan-1-one; 1-(3-(4-(3-fluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)ethan-1-one;1-(3-(4-(5-Fluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)ethan-1-one; 1-(3-(4-(3,5-Bis(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)ethan-1-one; 1-(3-(4-(4-Fluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)ethan-1-one; (4-(2-Fluoro-6-(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (4-(3-Fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(2,2,2-trifluoroethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; (5-(Cyclopropylmethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(3-Fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; Methyl 3-(4-(3-Fluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate; 3-(4-(3-Fluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carbonitrile; 1-(3-(4-(2-Chloro-5-fluorophenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)ethan-1-one; (4-(2-Fluoro-6-(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; tert-Butyl 2-(3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)acetate; tert-Butyl 3-(4-(3,5-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate; tert-Butyl 3-(4-(3,5-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylate; tert-Butyl 3-(4-(2-Fluoro-6-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate;tert-Butyl 3-(4-(5-fluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate; tert-Butyl 3-(4-(5-fluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylate; tert-Butyl 3-(4-(2-chloro-3-fluorophenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate; tert-Butyl 3-(4-(2-fluoro-6-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylate; tert-Butyl 3-(4-(3,5-bis(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate; tert-Butyl 3-(4-(3,5-bis(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylate; tert-Butyl 3-(4-(2-chloro-5-fluorophenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate; tert-Butyl 3-(4-(2-chloro-5-fluorophenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylate; tert-Butyl 3-(4-(4-fluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylate; tert-Butyl 3-(4-(3-fluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylate; tert-Butyl 3-(4-(3,4-difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate; tert-Butyl 3-(4-(3,4-difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylate; tert-Butyl 3-(4-(3-fluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate; tert-Butyl 3-(4-(4-fluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate; tert-Butyl 3-(4-(2-chloro-3-fluorophenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylate;(6,6-Dimethyl-1,4,6,7-tetrahydropyrano[4,3-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6,6-dioxido-1,4,5,7-tetrahydrothieno[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1,4,6,7-tetrahydropyrano[4,3-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1,4,5,7-tetrahydropyrano[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1-Methyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1-Methyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 1-Ethyl-N,N-dimethyl-3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxamide; (5-(2,2,2-Trifluoroethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-(2,2,2-Trifluoroethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-Chloro-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1H-Pyrazolo[3,4-b]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Chloro-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(Methylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 1-(3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one; (4,5,6,7-Tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Fluoro-1-(oxetan-3-yl)-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1-Ethyl-6-fluoro-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone;(6-Fluoro-1-isopropyl-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 1-(3-(4-Fluoro-4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one; (5-Fluoro-1-methyl-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Fluoro-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Methyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-(Methylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)ethan-1-one; (5-Fluoro-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-((Chloromethyl)sulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(2-Methoxyethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (4-Fluoro-4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)methanone; 1-(3-(4-(4-Fluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)ethan-1-one; (1-Ethyl-5-fluoro-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Fluoro-1-methyl-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[4,3-c]pyridin-6-one; 3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[3,4-c]pyridin-5-one; 6-Methyl-3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[3,4-c]pyridin-5-one;5-Methyl-3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[4,3-c]pyridin-6-one; (5,5-dioxido-1,4,6,7-tetrahydrothieno[4,3-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1-methyl-5-(methylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1-methyl-6-(methylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 1-(1-ethyl-3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)ethan-1-one; (5-(methoxymethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-(methoxymethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-methoxy-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(oxetan-3-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-isobutyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)propan-1-one; (5-ethyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 3-methyl-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)butan-1-one; 2-methyl-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)propan-1-one; 2,2-dimethyl-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)propan-1-one;(5-(isopropylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(isobutylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(ethylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-1H-indazole-5-carbonitrile; (7-chloro-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5,6-difluoro-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-(2-methoxyethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 3,3,3-trifluoro-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)propan-1-one; (5-(tert-butyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-isopropyl-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; N-methyl-3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxamide; N-methyl-3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxamide; (5-bromo-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxylic acid tert-butyl ester; 3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylic acid tert-butyl ester; (5-fluoro-1-isopropyl-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (7-fluoro-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone;(1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1H-pyrazolo[4,3-b]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-methoxy-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-fluoro-1-(oxetan-3-yl)-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1-ethyl-5-(methylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1-ethyl-6-(methylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 1-(1-methyl-3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one; 1-(1-methyl-3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)ethan-1-one; N,N-dimethyl-3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxamide; N,N-dimethyl-3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridine-6-carboxamide; (1-methyl-5,5-dioxido-1,4,6,7-tetrahydrothieno[4,3-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)(1,6,6-trimethyl-1,4,6,7-tetrahydropyrano[4,3-c]pyrazol-3-yl)methanone; (1-methyl-1,4,6,7-tetrahydropyrano[4,3-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 2-methyl-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)propan-1-one; (6-(isopropylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone;(6-(Ethylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)propan-1-one; 2-Methoxy-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one; 3,3,3-Trifluoro-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)propan-1-one; (1H-Indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1-Methyl-1H-indazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-(Oxetan-3-yl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-(tert-Butylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 2,2-Dimethyl-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)propan-1-one; (6-(tert-Butyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-(Isobutylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 3-Methyl-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6-yl)butan-1-one; (6-Isobutyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Isopropyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Ethyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone;(5-(tert-Butylsulfonyl)-4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; tert-Butyl 3-(4-fluoro-4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate; (4-Hydroxy-4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)(1-methyl-1H-indazol-3-yl)methanone; 1-(3-(4-Hydroxy-4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)ethan-1-one; 3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-N-methyl-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-neopentyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)methanone; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(oxetan-3-yl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)methanone; (5-(Cyclopropylmethyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(3,4-difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-ethyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)methanone; 1-(3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazol-5(1H)-yl)-3-methylbutan-1-one; 1-(3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazol-5(1H)-yl)-2-methylpropan-1-one; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-pyridinecarbonyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)methanone; 3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carbonitrile; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(2-methoxyethyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)methanone;(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(3,3,3-trifluoropropyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)methanone; (5-Benzoyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(3,4-difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; Methyl 3-(4-(3,4-difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)methanone; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(2,2,2-trifluoroethyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)methanone; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(pyrrolidine-1-carbonyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)methanone; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-isonicotinoyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)methanone; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-nicotinoyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)methanone; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(piperidine-1-carbonyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)methanone; (4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(5-(piperazine-1-carbonyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)methanone; 1-(3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazol-5(1H)-yl)propan-1-one; (5,5-Dioxido-4,6-dihydro-1H-thieno[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (4,6-Dihydro-1H-furo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1-Ethyl-5-(methylsulfonyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone;(1-Methyl-5-(methylsulfonyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1-Methyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 2-Methoxy-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazol-5(1H)-yl)ethan-1-one; (5-(2-Methoxyethyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 3,3,3-Trifluoro-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazol-5(1H)-yl)propan-1-one; (5-(Oxetan-3-yl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(Isobutylsulfonyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(Isopropylsulfonyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(Ethylsulfonyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; tert-Butyl 3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate; (5-Methyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(Methylsulfonyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 1-(3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazol-5(1H)-yl)ethan-1-one; (5-(tert-Butylsulfonyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(tert-Butyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone;(5-Isobutyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-Isopropyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-Ethyl-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; N-Methyl-3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide; 1-(1-Methyl-3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-yl)ethan-1-one; 2,2-Dimethyl-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-yl)propan-1-one; 3-Methyl-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-yl)butan-1-one; 2-Methyl-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-yl)propan-1-one; 1-(3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-yl)propan-1-one; N,N-Dimethyl-3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide; (5-(2,2,2-Trifluoroethyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(Methoxymethyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1,4,5,6-Tetrahydropyrrolo[3,4-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 4-(3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazole-5-carbonyl)piperazine-1-carboxylic acid tert-butyl ester; 3-(4-(3,4-Difluoro-2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylic acid tert-butyl ester; (5,5-Dioxido-4,6,7,8-tetrahydro-1H-thia; (thiepino)[4,3-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (4,6,7,8-tetrahydro-1H-oxa (oxepino)[4,3-c]pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; N-methyl-3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6,7,8-tetrahydropyrazolo[4,3-c]azepin -5(1H)-carboxamide; (5-(2,2,2-trifluoroethyl)-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(tert-butylsulfonyl)-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 2,2-dimethyl-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6,7,8-tetrahydropyrazolo[4,3-c]azepin -5(1H)-yl)propan-1-one; (5-(tert-butyl)-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(isobutylsulfonyl)-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 3-methyl-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6,7,8-tetrahydropyrazolo[4,3-c]azepin -5(1H)-yl)butan-1-one; (5-isobutyl-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(isopropylsulfonyl)-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 2-methyl-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6,7,8-tetrahydropyrazolo[4,3-c]azepin -5(1H)-yl)propan-1-one; (1-ethyl-5-(methylsulfonyl)-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 1-(1-methyl-3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6,7,8-tetrahydropyrazolo[4,3-c]azepin -5(1H)-yl)ethan-1-one; (5-(methoxymethyl)-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-methyl-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-isopropyl-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(ethylsulfonyl)-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6,7,8-tetrahydropyrazolo[4,3-c]azepin -5(1H)-yl)propan-1-one; (5-ethyl-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(methylsulfonyl)-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6,7,8-tetrahydropyrazolo[4,3-c]azepin -5(1H)-yl)ethan-1-one; (1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1-methyl-5-(methylsulfonyl)-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1-methyl-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepin -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; N,N-dimethyl-3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6,7,8-tetrahydropyrazolo[4,3-c]azepine -5(1H)-carboxamide; (5-(2-methoxyethyl)-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepine -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 2-methoxy-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6,7,8-tetrahydropyrazolo[4,3-c]azepine -5(1H)-yl)ethan-1-one; 3,3,3-trifluoro-1-(3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6,7,8-tetrahydropyrazolo[4,3-c]azepine -5(1H)-yl)propan-1-one; (5-(oxetan-3-yl)-1,4,5,6,7,8-hexahydropyrazolo[4,3-c]azepine -3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 3-(4-(2-(trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,6,7,8-tetrahydropyrazolo[4,3-c]azepine tert-Butyl -5(1H)-formate; (6-(trifluoromethyl)imidazo[1,2-b]pyridazin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-fluoroimidazo[1,2-b]pyridazin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-(pyrrolidin-1-yl)imidazo[1,2-b]pyridazin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-cyclopropylimidazo[1,2-b]pyridazin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-methoxyimidazo[1,2-b]pyridazin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-methylimidazo[1,2-b]pyridazin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-chloroimidazo[1,2-b]pyridazin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; imidazo[1,2-b]pyridazin-2-yl(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-chloro-2-methylimidazo[1,2-b]pyridazin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1H-benzo[d]imidazol-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1H-imidazo[4,5-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (4-(5-fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; (4-(3-fluoro-2-(trifluoromethyl)phenyl)piperidin-1-yl)(6-(2-methoxyethyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-3-yl)methanone; 6-methyl-2-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)pyrimidine-4-carboxylic acid; methyl 6-methyl-2-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)pyrimidine-4-carboxylate; N-(cyclopropylsulfonyl)-2-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)benzamide; N-(phenylsulfonyl)-2-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)benzamide; N-(methylsulfonyl)-2-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)benzamide; 3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)benzamide; 2-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)benzamide; 4-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)benzoic acid; 3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-carbonyl)benzoic acid;2-(4-(2-(Trifluoromethyl)phenyl)piperidine-1-carbonyl)benzoic acid; 4-(4-(2-(tert-Butyl)phenyl)piperidine-1-carbonyl)benzoic acid; 2-(4-(2-(tert-Butyl)phenyl)piperidine-1-carbonyl)benzoic acid; 3-(4-(2-(tert-Butyl)phenyl)piperidine-1-carbonyl)benzoic acid; 4-(4-(2-(Trifluoromethyl)phenyl)piperidine-1-carbonyl)benzamide; 1-(3-(4-(2-(Trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,7-dihydroisothiazolo[5,4-c]pyridin-6(5H)-yl)ethan-1-one; (4,5,6,7-Tetrahydroisothiazolo[5,4-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (4,5,6,7-Tetrahydroisothiazolo[4,5-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 1-(3-(4-(2-(Trifluoromethyl)phenyl)piperidine-1-carbonyl)-6,7-dihydroisoxazolo[4,5-c]pyridin-5(4H)-yl)ethan-1-one; 1-(3-(4-(2-(Trifluoromethyl)phenyl)piperidine-1-carbonyl)-4,7-dihydroisoxazolo[5,4-c]pyridin-6(5H)-yl)ethan-1-one; (4,5,6,7-Tetrahydroisoxazolo[4,5-c]pyridin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; Benzo[c]isothiazol-3-yl(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; Benzo[d]thiazol-2-yl(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; Benzo[d]isoxazol-3-yl(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 1-(3-(4-(2-(Trifluoromethyl)phenyl)piperidine-1-carbonyl)-6,7-dihydroisothiazolo[4,5-c]pyridin-5(4H)-yl)ethan-1-one; Benzo[d]oxazol-2-yl(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (3-Methyloxetan-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; Oxetan-3-yl(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 2-(2-Hydroxyphenyl)-1-(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)ethan-1-one; (4-(2-(tert-Butyl)phenyl)piperidin-1-yl)(tetrahydrothiophen-2-yl)methanone; rac-(2R,3R)-2-(4-(2-(tert-Butyl)phenyl)piperidine-1-carbonyl)-3-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester; rac-tert-Butyl (2R,4R)-2-(4-(2-(tert-Butyl)phenyl)piperidine-1-carbonyl)-4-hydroxypyrrolidine-1-carboxylate 2-(2-Oxo-2-(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)ethyl)phenylsulfamate;(4-(2-(tert-Butyl)phenyl)piperidin-1-yl)(1,1-dioxidotetrahydrothiophen-2-yl)methanone; rac-(4-(2-(tert-Butyl)phenyl)piperidin-1-yl)((2R,3R)-3-hydroxypyrrolidin-2-yl)methanone; rac-(4-(2-(tert-Butyl)phenyl)piperidin-1-yl)((2R,4R)-4-hydroxypyrrolidin-2-yl)methanone; rac-(R)-1-(2-(4-(2-(tert-Butyl)phenyl)piperidine-1-carbonyl)pyrrolidin-1-yl)ethan-1-one; (6-Bromo-1H-pyrrolo[3,2-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-Morpholino-1H-pyrrolo[3,2-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-(1H-Imidazol-1-yl)-1H-pyrrolo[3,2-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-Chloro-1H-pyrrolo[3,2-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-Fluoro-1H-pyrrolo[3,2-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (5-Methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Methoxy-1H-pyrrolo[3,2-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; Imidazo[1,2-a]pyridin-2-yl(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Chloro-2-methylimidazo[1,2-b]pyridazin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; Imidazo[1,2-b]pyridazin-6-yl(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1H-Pyrrolo[2,3-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1H-Pyrrolo[3,2-c]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Chloro-1H-pyrrolo[3,2-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Morpholino-1H-pyrrolo[3,2-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-(1H-Imidazol-1-yl)-1H-pyrrolo[3,2-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1-Methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone;(5-Methoxy-1H-pyrrolo[3,2-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Fluoro-1H-pyrrolo[3,2-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1H-Imidazo[4,5-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Methyl-1H-pyrrolo[3,2-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1H-Indol-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1H-Pyrrolo[3,2-b]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1H-Pyrrolo[2,3-c]pyridin-2-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1H-Pyrazol-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (1H-1,2,3-Triazol-5-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; Pyrazin-2-yl(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Methoxypyridazin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Methylpyridazin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (4-Methyl-1,2,3-thiadiazol-5-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; (6-Chloropyridazin-3-yl)(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; Pyridazin-3-yl(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; Pyridazin-4-yl(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)methanone; 4-(2-(Trifluoromethyl)phenyl)piperidine-1-carboxylic acid; or 3-Oxo-3-(4-(2-(trifluoromethyl)phenyl)piperidin-1-yl)propanoic acid.;

[0275] Some embodiments provided herein describe RBP4 inhibitors or pharmaceutically acceptable salts, solvates, polymorphs, prodrugs, metabolites, N-oxides, stereoisomers or isomers thereof for the treatment of age-related macular degeneration and having a structure of formula (II):

[0276]

[0277] Wherein:

[0278] Ring A B is optionally further substituted benzene;

[0279] R B 1is an optionally substituted branched C 3-6 alkyl;

[0280] X B 1 is O, S, SO, SO 2 or NH;

[0281] X B 2 is a bond or C 1 -C 3 alkylene;

[0282] Ring B B is azetidine, pyrrolidine or piperidine;

[0283] X B 3 is CO or SO 2 ;

[0284] R B 2 is a substituent; or

[0285] its pharmaceutically acceptable salts.

[0286] In some embodiments, the compound of formula (II) has the structure

[0287]

[0288] wherein:

[0289] Ring A B is an optionally further substituted benzene;

[0290] R B 1 is an optionally substituted branched C 3 -C 6 alkyl

[0291] X B 1 is O, S, SO, SO 2 or NH;

[0292] X B 2 is a bond or C 1 -C 3 alkylene;

[0293] Ring B B is azetidine or piperidine;

[0294] X B 3 is CO or SO 2 ;

[0295] RB 2 is an optionally substituted hydrocarbon group, an optionally substituted heterocyclic group, an optionally substituted hydroxyl group, an optionally substituted mercapto group, a cyano group, a nitro group, an acyl group or a halogen atom; or

[0296] its pharmaceutically acceptable salt.

[0297] In some embodiments, the compound of formula (II) is 4-(3-(2-tert-butylphenoxy)azetidin-1-yl)-4-oxobutanoic acid, 3-{3-[(2-tert-butyl-4-fluorophenoxy)methyl]azetidin-1-yl}-3-oxopropanoic acid, 2-{[3-(2-tert-butyl-4-chlorophenoxy)azetidin-1-yl]carbonyl}pyridine, 4-[3-(2-tert-butyl-4-chlorophenoxy)azetidin-1-yl]-4-oxobutanoic acid, {3-[(2-tert-butyl-4-chlorophenoxy)methyl]azetidin-1-yl}(oxo)acetic acid, {3-[(2-tert-butylphenoxy)methyl]azetidin-1-yl}(oxo)acetic acid, 3-{3-[(2-tert-butylphenoxy)methyl]azetidin-1-yl}-3-oxopropanoic acid, {4-[(2-tert-butyl-4-chlorophenoxy)methyl]piperidin-1-yl}(oxo)acetic acid, [4-(2-tert-butylphenoxy)piperidin-1-yl](oxo)acetic acid or {4-[(2-tert-butylphenoxy)methyl]piperidin-1-yl}(oxo)acetic acid, or its pharmaceutically acceptable salt.

[0298] Some embodiments provided herein describe an RBP4 inhibitor or its pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer for treating age-related macular degeneration and having the structure of formula (III):

[0299]

[0300] Wherein:

[0301] Ring A C is an optionally further substituted benzene ring;

[0302] Ring B C is an optionally further substituted piperazine ring; and

[0303] R C is a substituent; or

[0304] its pharmaceutically acceptable salt.

[0305] In some embodiments, the compound of formula (III) has the structure

[0306]

[0307] Wherein:

[0308] Ring A C is a benzene ring optionally substituted with 1 to 3 substituents selected from: (a) a halogen atom and (b) C 1-6 alkyl;

[0309] Ring B C is a piperazine ring optionally substituted with 1 to 3 substituents selected from: (a) a halogen atom, (b) C optionally substituted with 1 to 3 halogen atoms 1 -C 6 alkyl and (c) C optionally substituted with 1 to 3 halogen atoms 1 -C 6 alkoxy; and

[0310] R C is (1) optionally substituted C 1 -C 10 alkyl, (2) optionally substituted C 6 -C 14 aryl, (3) optionally substituted 5- or 6-membered aromatic heterocyclic group, (4) optionally substituted amino group, (5) optionally substituted carboxyl group or (6) optionally substituted carbamoyl group; or

[0311] a pharmaceutically acceptable salt thereof.

[0312] In some embodiments, the compound of formula (III) is N-{[4-(2-tert-butylphenyl)piperazin-1-yl]carbonyl}glycine, 3-[4-(2-tert-butylphenyl)piperazin-1-yl]-3-oxopropanoic acid, [4-(2-tert-butyl-4-chlorophenyl)piperazin-1-yl](oxo)acetic acid, 5-{2-[4-(2-tert-butylphenyl)piperazin-1-yl]-2-oxoethyl}imidazolidine-2,4-dione, [(5-{[4-(2-tert-butylphenyl)piperazin-1-yl]carbonyl}isoxazol-3-yl)oxy]acetic acid or a pharmaceutically acceptable salt thereof.

[0313] Some embodiments provided herein describe an RBP4 inhibitor or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof for the treatment of age-related macular degeneration, having the structure of formula (IV):

[0314]

[0315] Wherein:

[0316] Ring A D is a 5-membered non-aromatic heterocycle, which is optionally further substituted with one substituent;

[0317] Ring B D is an optionally further substituted benzene ring; and

[0318] X D is a bond, O, CH 2 O, OCH 2 , CH 2 , (CH 2 ) 2 , S, CH 2 S, SCH 2 , S(O), CH 2 S(O), S(O)CH 2 , S(O) 2 , CH 2 S(O) 2 or S(O) 2 CH 2 ; or

[0319] its pharmaceutically acceptable salts.

[0320] In some embodiments, the compound of formula (IV) has the structure

[0321]

[0322] wherein:

[0323] Ring A D is a 5-membered non-aromatic heterocycle, which is optionally further substituted by an oxo group;

[0324] Ring B D is a benzene ring, which is optionally further substituted by 1 to 4 substituents; and

[0325] X D is O, CH 2 O, OCH 2 , CH 2 , (CH 2 ) 2 , S, CH 2 S, SCH 2 , S(O), CH 2 S(O), S(O)CH 2 , S(O) 2 , CH 2 S(O) 2 or S(O) 2 CH 2 ; or

[0326] its pharmaceutically acceptable salts.

[0327] In some embodiments, the compound of formula (IV) is ({(3S)-1-[3,5-bis(trifluoromethyl)phenyl]pyrrolidin-3-yl}oxy)acetic acid, ({1-[4-chloro-3-(trifluoromethyl)phenyl]pyrrolidin-3-yl}sulfanyl)acetic acid, 3-{(2R,5S)-5-[3,5-bis(trifluoromethyl)phenyl]tetrahydrofuran-2-yl}propanoic acid, or a pharmaceutically acceptable salt thereof.

[0328] Some embodiments provided herein describe an RBP4 inhibitor or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof for treating age-related macular degeneration and having the structure of formula (V):

[0329]

[0330] Wherein:

[0331] Ring A E is an optionally further substituted monocyclic nitrogen-containing aromatic heterocycle;

[0332] X E is CH 2 or O; and

[0333] R E is a hydrogen atom or C 1 -C 6 alkyl; or

[0334] its pharmaceutically acceptable salt.

[0335] In some embodiments, the compound of formula (V) has the structure

[0336]

[0337] Wherein:

[0338] Ring A E is a pyrazole ring, pyridine ring, oxazole ring, imidazole ring or pyrimidine ring;

[0339] X E is S, an optionally substituted alkylene or O; and

[0340] R E is a hydrogen atom or C 1 -C 6 alkyl; or

[0341] its pharmaceutically acceptable salt.

[0342] In some embodiments, the compound of formula (V) is ((4-(3,5-bis(trifluoromethyl)phenyl)-1,3-oxazol-2-yl)thio)acetic acid, ethyl ((6-(3,5-bis(trifluoromethyl)phenyl)pyridin-3-yl)thio)acetate, ((6-(3,5-bis(trifluoromethyl)phenyl)pyridin-3-yl)thio)acetic acid, or 3-(3-(3,5-bis(trifluoromethyl)phenyl)-1H-pyrazol-1-yl)butyric acid.

[0343] In some embodiments, the RBP4 inhibitor is the RBP4 inhibitor provided in Table B.

[0344] Table B: RBP4 inhibitors

[0345]

[0346]

[0347]

[0348]

[0349] Preparation of the compound

[0350] The compounds used in the chemical reactions described herein are prepared according to organic synthesis techniques known to those skilled in the art, starting from commercially available chemicals and / or compounds described in the chemical literature. "Commercially available chemicals" are obtained from standard commercial sources, including Acros Organics (Pittsburgh, PA), Aldrich Chemical (Milwaukee, WI, including Sigma Chemical and Fluka), Apin Chemicals Ltd. (Milton Park, UK), Avocado Research (Lancashire, U.K.), BDH Inc. (Toronto, Canada), Bionet (Cornwall, U.K.), Chemservice Inc. (West Chester, PA), Crescent Chemical Co. (Hauppauge, NY), Eastman Organic Chemicals, Eastman Kodak Company (Rochester, NY), Fisher Scientific Co. (Pittsburgh, PA), Fisons Chemicals (Leicestershire, UK), Frontier Scientific (Logan, UT), ICN Biomedicals, Inc. (Costa Mesa, CA), Key Organics (Cornwall, U.K.), Lancaster Synthesis (Windham, NH), Maybridge Chemical Co. Ltd. (Cornwall, U.K.), Parish Chemical Co. (Orem, UT), Pfaltz & Bauer, Inc. (Waterbury, CN), Polyorganix (Houston, TX), Pierce Chemical Co. (Rockford, IL), Riedel de Haen AG (Hanover, Germany), Spectrum Quality Product, Inc. (New Brunswick, NJ), TCI America (Portland, OR), Trans World Chemicals, Inc. (Rockville, MD), and Wako Chemicals USA, Inc. (Richmond, VA).

[0351] Suitable reference books and papers that describe in detail the synthesis of the reactants that can be used to prepare the compounds described herein or provide references to articles describing such preparations include, for example, "Synthetic Organic Chemistry", John Wiley & Sons, Inc., New York; S.R. Sandler et al., "Organic Functional Group Preparations," 2nd ed., Academic Press, New York, 1983; H.O. House, "Modern Synthetic Reactions," 2nd ed., W.A. Benjamin, Inc. Menlo Park, Calif. 1972; T.L. Gilchrist, "Heterocyclic Chemistry," 2nd ed., John Wiley & Sons, New York, 1992; J. March, "Advanced Organic Chemistry: Reactions, Mechanisms and Structure," 4th ed., Wiley-Interscience, New York, 1992. Other suitable reference books and papers that describe in detail the synthesis of the reactants that can be used to prepare the compounds described herein or provide references to articles describing such preparations include, for example, Fuhrhop, J and Penzlin G. "Organic Synthesis: Concepts, Methods, Starting Materials", Second Revised and Enlarged Edition (1994) John Wiley & Sons ISBN: 3-527-29074-5; Hoffman, R.V. "Organic Chemistry, An Intermediate Text" (1996) Oxford University Press, ISBN 0-19-509618-5; Larock, R.C. "Comprehensive Organic Transformations: A Guide to Functional Group Preparations" 2nd ed. (1999) Wiley-VCH, ISBN: 0-471-19031-4; March, J. "Advanced Organic Chemistry: Reactions, Mechanisms, and Structure" 4th ed. (1992) John Wiley & Sons, ISBN: 0-471-60180-2; Otera, J.(Editor) "Modern Carbonyl Chemistry" (2000) Wiley-VCH, ISBN: 3-527-29871-1; Patai, S. "Patai's 1992 Guide to the Chemistry of Functional Groups" (1992) Interscience ISBN: 0-471-93022-9; Solomons, T.W.G. "Organic Chemistry" 7th Edition (2000) John Wiley & Sons, ISBN: 0-471-19095-0; Stowell, J.C., "Intermediate Organic Chemistry" 2nd Edition (1993) Wiley-Interscience, ISBN: 0-471-57456-2; "Industrial Organic Chemicals: Starting Materials and Intermediates: An Ullmann's Encyclopedia" (1999) John Wiley & Sons, ISBN: 3-527-29645-X, 8 volumes; "Organic Reactions" (1942 - 2000) John Wiley & Sons, over 55 volumes and "Chemistry of Functional Groups" John Wiley & Sons, 73 volumes.

[0352] Alternatively, specific and similar reactants can be identified through indexes of known chemicals and reactions compiled by the Chemical Abstract Service of the American Chemical Society, which are available in most public and university libraries, or through online databases (for details, contact the American Chemical Society in Washington, D.C.). Chemicals that are known but not listed in commercial catalogs are optionally prepared by conventional chemical synthesis companies, and many standard chemical supply companies (e.g., those listed above) offer conventional synthesis services. References for the preparation and selection of pharmaceutical salts of the heterocyclic RBP4 inhibitory compounds described herein are P.H. Stahl & C.G. Wermuth "Handbook of Pharmaceutical Salts", Verlag Helvetica Chimica Acta, Zurich, 2002.

[0353] Retinol Binding Protein 4 (RBP4)

[0354] Retinol binding protein 4 (RBP4) is the only retinol transport protein in the blood and is secreted by adipocytes and the liver. Reducing RBP4 levels can reduce the accumulation of lipofuscin, and the accumulation of lipofuscin can lead to vision loss in diseases such as STGD, AMD, dry (atrophic) age-related macular degeneration. In some cases, reducing RBP4 reduces the accumulation of lipofuscin in the retina. In some embodiments, the compounds and formulations described herein reduce serum or plasma RBP4, thereby delaying or preventing vision loss due to excessive accumulation of lipofuscin in the retina. In some embodiments, the compounds and formulations described herein reduce serum or plasma RBP4, thereby delaying or preventing vision loss due to age-related macular degeneration.

[0355] In some embodiments, 48 hours after administration of an RBP4 inhibitor, its pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer, the serum or plasma level of RBP4 is reduced by at least 30% from baseline. In some embodiments, 48 hours after administration of an RBP4 inhibitor, its pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer, the serum or plasma level of RBP4 is reduced by at least 40% from baseline. In some embodiments, 48 hours after administration of an RBP4 inhibitor, its pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer, the serum or plasma level of RBP4 is reduced by at least 50% from baseline. In other embodiments, 48 hours after administration of an RBP4 inhibitor, its pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer, the serum or plasma level of RBP4 is reduced by at least 65% from baseline. In certain embodiments, 48 hours after administration of an RBP4 inhibitor, its pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer, the serum or plasma level of RBP4 is reduced by at least 80% from baseline. In some embodiments, 48 hours after administration of an RBP4 inhibitor, its pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer, the serum or plasma level of RBP4 is reduced by at least 85% from baseline.

[0356] In some embodiments, 36 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 30% from baseline. In some embodiments, 36 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 40% from baseline. In some embodiments, 36 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 50% from baseline. In other embodiments, 36 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 65% from baseline. In certain embodiments, 36 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 80% from baseline. In some embodiments, 36 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 85% from baseline.

[0357] In some embodiments, 24 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 30% from baseline. In some embodiments, 24 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 40% from baseline. In some embodiments, 24 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 50% from baseline. In other embodiments, 24 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 65% from baseline. In certain embodiments, 24 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 80% from baseline. In some embodiments, 24 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 85% from baseline.

[0358] In some embodiments, 12 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 30% from baseline. In some embodiments, 12 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 40% from baseline. In some embodiments, 12 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 50% from baseline. In other embodiments, 12 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 65% from baseline. In certain embodiments, 12 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 80% from baseline. In some embodiments, 12 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 85% from baseline.

[0359] In some embodiments, 6 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 20% from baseline. In some embodiments, 6 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 25% from baseline. In some embodiments, 6 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 30% from baseline. In some embodiments, 6 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 40% from baseline. In some embodiments, 6 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 50% from baseline. In other embodiments, 6 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 65% from baseline. In certain embodiments, 6 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 80% from baseline. In some embodiments, 6 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer or isomer thereof, the serum or plasma level of RBP4 is reduced by at least 85% from baseline.

[0360] In some embodiments, the serum or plasma level of RBP4 is reduced by at least 1 mg / dL 48 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In other embodiments, the serum or plasma level of RBP4 is reduced by at least 2 mg / dL 48 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In some embodiments, the serum or plasma level of RBP4 is reduced by at least 5 mg / dL 48 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In certain embodiments, the serum or plasma level of RBP4 is reduced by at least 10 mg / dL 48 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In some embodiments, the serum or plasma level of RBP4 is reduced by at least 15 mg / dL 48 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof.

[0361] In some embodiments, the serum or plasma level of RBP4 is reduced by at least 1 mg / dL 36 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In other embodiments, the serum or plasma level of RBP4 is reduced by at least 2 mg / dL 36 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In some embodiments, the serum or plasma level of RBP4 is reduced by at least 5 mg / dL 36 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In certain embodiments, the serum or plasma level of RBP4 is reduced by at least 10 mg / dL 36 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In some embodiments, the serum or plasma level of RBP4 is reduced by at least 15 mg / dL 36 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof.

[0362] In some embodiments, the serum or plasma level of RBP4 is reduced by at least 1 mg / dL 24 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In other embodiments, the serum or plasma level of RBP4 is reduced by at least 2 mg / dL 24 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In some embodiments, the serum or plasma level of RBP4 is reduced by at least 5 mg / dL 24 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In certain embodiments, the serum or plasma level of RBP4 is reduced by at least 10 mg / dL 24 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In some embodiments, the serum or plasma level of RBP4 is reduced by at least 15 mg / dL 24 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof.

[0363] In some embodiments, the serum or plasma level of RBP4 is reduced by at least 1 mg / dL 12 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In other embodiments, the serum or plasma level of RBP4 is reduced by at least 2 mg / dL 12 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In some embodiments, the serum or plasma level of RBP4 is reduced by at least 5 mg / dL 12 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In certain embodiments, the serum or plasma level of RBP4 is reduced by at least 10 mg / dL 12 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In some embodiments, the serum or plasma level of RBP4 is reduced by at least 15 mg / dL 12 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof.

[0364] In some embodiments, the serum or plasma level of RBP4 is reduced by at least 1 mg / dL 6 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In other embodiments, the serum or plasma level of RBP4 is reduced by at least 2 mg / dL 6 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In some embodiments, the serum or plasma level of RBP4 is reduced by at least 5 mg / dL 6 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In certain embodiments, the serum or plasma level of RBP4 is reduced by at least 10 mg / dL 6 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof. In some embodiments, the serum or plasma level of RBP4 is reduced by at least 15 mg / dL 6 hours after administration of an RBP4 inhibitor, a pharmaceutically acceptable salt, solvate, polymorph, prodrug, metabolite, N-oxide, stereoisomer, or isomer thereof.

[0365] Method of treatment

[0366] In some embodiments, the compounds disclosed herein are used to treat or ameliorate a disease associated with an altered RBP4 pathway, such as AMD or STGD, when administered to a subject in need thereof. In some cases, the compounds disclosed herein are used to treat or ameliorate the effects of a disease associated with an altered RBP4 pathway, such as age-related macular degeneration or STGD, when administered to a subject in need thereof.

[0367] Age-related macular degeneration

[0368] Age-related macular degeneration (AMD) is an eye condition common in people over 50 years of age and is a leading cause of vision loss. It damages the macula, a small area near the center of the retina and the part of the eye needed for sharp central vision. As AMD progresses, it is common to have blurry areas near the center of vision. Over time, the blurry areas may grow larger, and the subject may develop blind spots in their central vision.

[0369] Some embodiments provided herein describe the use of the RBP4 inhibitors described herein for treating AMD in a subject in need thereof. In some embodiments, the RBP4 inhibitor inhibits AMD. In certain embodiments, the RBP4 inhibitor prevents the development of AMD or its clinical symptoms. In certain embodiments, the RBP4 inhibitor alleviates the development of AMD or its clinical symptoms. In certain embodiments, the RBP4 inhibitor relieves AMD in the subject. In certain embodiments, the RBP4 inhibitor causes regression, reversal, or improvement of AMD. In certain embodiments, the RBP4 inhibitor reduces the number, frequency, duration, or severity of the clinical symptoms of AMD.

[0370] In some embodiments, the RBP4 inhibitor is used prophylactically. In certain embodiments, the RBP4 inhibitor is used to prevent or reduce the risk of developing AMD. In certain embodiments, the RBP4 inhibitor prevents the clinical symptoms of AMD from developing in a subject who may be predisposed to AMD but has not yet experienced or exhibited symptoms of AMD.

[0371] Dry (atrophic) age-related macular degeneration

[0372] Approximately 85% to 90% of cases of macular degeneration are of the "dry" (atrophic) type. It is estimated that 62.9 million people worldwide have this type of AMD; 8 million of them are Americans. Due to increasing life expectancy and current demographics, this number is expected to triple by 2020. There is currently no FDA-approved treatment for dry AMD. Given the lack of treatment and high prevalence, the development of drugs for dry AMD is crucial. Clinically, atrophic AMD represents a slowly progressive neurodegenerative disease in which specialized neurons (rod and cone photoreceptor cells) die in the central part of the retina called the macula. Histopathological and clinical imaging studies have shown that photoreceptor degeneration in dry AMD is triggered by abnormalities in the retinal pigment epithelium (RPE) located beneath the photoreceptors and providing key metabolic support for these photoreceptor neurons. Experimental and clinical data have shown that the excessive accumulation of cytotoxic autofluorescent lipid-protein-retinoid aggregates (lipofuscin) in the RPE is the major inducer of dry AMD. The major cytotoxic component of RPE lipofuscin is the pyridinium bisretinoid A2E. Other cytotoxic bisretinoids are isoA2E, atRALdi-PE, and A2-DHP-PE. The formation of A2E and other lipofuscin bisretinoids such as A2-DHP-PE (A2-dihydropyridinium-phosphatidylethanolamine) and atRALdi-PE (all-trans-retinal dimer-phosphatidylethanolamine) starts in a non-enzymatic manner in photoreceptors and can be considered a byproduct of the normal functional visual cycle.

[0373] Some embodiments provided herein describe the use of the RBP4 inhibitors described herein for treating dry (atrophic) AMD in a subject in need thereof. In some embodiments, the RBP4 inhibitor inhibits dry (atrophic) AMD. In certain embodiments, the RBP4 inhibitor prevents the development of dry (atrophic) AMD or its clinical symptoms. In certain embodiments, the RBP4 inhibitor reduces the development of dry (atrophic) AMD or its clinical symptoms. In certain embodiments, the RBP4 inhibitor alleviates dry (atrophic) AMD in a subject. In certain embodiments, the RBP4 inhibitor causes regression, reversal, or improvement of dry (atrophic) AMD. In certain embodiments, the RBP4 inhibitor reduces the number, frequency, duration, or severity of the clinical symptoms of dry (atrophic) AMD.

[0374] In some embodiments, the RBP4 inhibitor is used prophylactically. In certain embodiments, the RBP4 inhibitor is used to prevent or reduce the risk of developing dry (atrophic) AMD. In certain embodiments, the RBP4 inhibitor prevents the clinical symptoms of dry (atrophic) AMD from developing in a subject who may be predisposed to dry (atrophic) AMD but has not yet experienced or manifested symptoms of dry (atrophic) AMD.

[0375] Pharmaceutical composition

[0376] In certain embodiments, the RBP4 inhibitory compound as described herein is administered in pure chemical form. In other embodiments, the RBP4 inhibitory compounds described herein are combined with a pharmaceutically suitable or acceptable carrier (also referred to herein as a pharmaceutically suitable (or acceptable) excipient, a physiologically suitable (or acceptable) excipient, or a physiologically suitable (or acceptable) carrier), which is selected based on the chosen route of administration and standard pharmaceutical practices such as those described in Remington: The Science and Practice of Pharmacy (Gennaro, 21st ed. Mack Pub. Co., Easton, PA (2005)).

[0377] Provided herein is a pharmaceutical composition comprising at least one RBP4 inhibitory compound or a stereoisomer, pharmaceutically acceptable salt, or N-oxide thereof, and one or more pharmaceutically acceptable carriers. A carrier is acceptable or suitable if it is compatible with the other ingredients of the composition and not harmful to the recipient (i.e., the subject or patient) of the composition.

[0378] One embodiment provides a pharmaceutical composition comprising an RBP4 inhibitor or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. In one embodiment, the pharmaceutical composition is provided in a dosage form for oral administration, which comprises a compound provided herein and one or more pharmaceutically acceptable excipients or carriers.

[0379] In certain embodiments, the RBP4 inhibitory compound is substantially pure in that it contains less than about 5% or less than about 1% or less than about 0.1% of other small organic molecules, such as unreacted intermediates or synthetic by-products generated in one or more steps of the synthetic method.

[0380] Suitable oral dosage forms include, for example, tablets, pills, capsules, or hard or soft gelatin, methylcellulose, or other suitable materials that are readily soluble in the digestive tract. In some embodiments, suitable non-toxic solid carriers are used, including, for example, pharmaceutical grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose, magnesium carbonate, etc. (See, for example, Remington: The Science and Practice of Pharmacy (Gennaro, 21st Edition Mack Pub. Co., Easton, PA (2005)).

[0381] In some embodiments, the pharmaceutical composition provided herein is formulated as a tablet, capsule, powder, or liquid form for oral administration. In some embodiments, the tablet contains a solid carrier or adjuvant. Liquid pharmaceutical compositions generally contain a liquid carrier, such as water, petroleum, animal or vegetable oils, mineral oil, or synthetic oils. In some embodiments, a physiological saline solution, dextrose, or other sugar solution, or ethylene glycol is optionally included. In some embodiments, the capsule contains a solid carrier, such as gelatin.

[0382] In another embodiment, the pharmaceutical composition is provided in a dosage form for parenteral administration, which comprises a compound provided herein and one or more pharmaceutically acceptable excipients or carriers. When the pharmaceutical composition is formulated for intravenous, cutaneous, or subcutaneous injection, the active ingredient is in the form of a parenterally acceptable aqueous solution that is pyrogen-free and has a suitable pH, isotonicity, and stability. Persons skilled in the relevant art can ...

Claims

1. A method for treating a subject suffering from an eye disease, the method comprising: (a) administering a treatment to the subject; (b) determining whether the treatment is therapeutically effective for treating the subject based on a composite biomarker; (c) predicting whether the prognostic potential of the treatment is improved based on the composite biomarker; wherein the composite biomarker is calculated based on at least two variables, and at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof.

2. A method for treating a subject suffering from an eye disease, the method comprising: (a) administering a treatment to the subject; (b) performing measurements on the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (c) calculating the at least two variables to generate a composite biomarker; and (d) determining whether the treatment is therapeutically effective for treating the subject based on the composite biomarker.

3. A method for treating a subject suffering from an eye disease, the method comprising: (a) administering a treatment to the subject; (b) performing a first measurement on the subject at a first time point to obtain a first set of at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (c) calculating the at least two variables of the first set to generate a first composite biomarker; (d) performing a second measurement on the subject at a second time point to obtain a second set of the at least two variables; (e) calculating the at least two variables of the second set to generate a second composite biomarker; and (f) determining whether the treatment is therapeutically effective for treating the subject based on the first and second composite biomarkers.

4. A method for treating a subject suffering from an eye disease, the method comprising: (a) administering a treatment to the subject; (b) performing measurements on the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; and (c) calculating the at least two variables to generate a composite biomarker, wherein when the composite biomarker exceeds a threshold, the treatment is therapeutically effective for treating the subject, and when the composite biomarker does not exceed the threshold, the treatment is not therapeutically effective for treating the subject.

5. A method for treating a subject suffering from an eye disease, the method comprising: (a) administering a first dose of treatment to the subject; (b) performing measurements on the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (c) calculating the at least two variables to generate a composite biomarker; (d) determining whether the treatment with the first dose is therapeutically effective for treating the subject based on the composite biomarker; and (e) administering a second dose of the treatment to the subject, wherein the second dose is different from the first dose when it is determined that the first dose is not therapeutically effective for treating the subject, and wherein the second dose is substantially the same as the first dose when it is determined that the first dose is therapeutically effective for treating the subject.

6. A method of treating a subject having an eye disease, the method comprising: (a) performing a first determination on the subject to obtain a first set of at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) calculating the at least two variables of the first set to produce a first composite biomarker; (c) administering a treatment to the subject; (d) performing a second determination on the subject to obtain a second set of the at least two variables; (e) calculating the at least two variables of the second set to produce a second composite biomarker; and (f) determining whether the treatment is therapeutically effective for treating the subject based on the first and second composite biomarkers.

7. A method of treating a subject having an eye disease, the method comprising: (a) performing a determination on the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) calculating the at least two variables to produce a composite biomarker; (c) selecting a treatment for the subject based on the composite biomarker; and (d) administering the treatment to the subject.

8. A method of treating a subject having an eye disease, the method comprising: (a) performing a first determination on the subject to obtain a first set of at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) calculating the at least two variables of the first set to produce a first composite biomarker; (c) selecting a treatment for the subject based on the first composite biomarker; (d) administering the treatment to the subject; (e) performing a second determination on the subject to obtain a second set of the at least two variables; (f) calculating the at least two variables of the second set to produce a second composite biomarker; and (g) determining whether the treatment is therapeutically effective for treating the subject based on the first and second composite biomarkers.

9. A method of treating a subject having an eye disease, the method comprising: (a) performing a determination on the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) calculating the at least two variables to produce a composite biomarker; (c) Select a treatment for the subject when the composite biomarker exceeds a threshold, or optionally, do not select the treatment for the subject when the composite biomarker does not exceed the threshold; and (d) Administer the treatment to the subject when the treatment is selected for the subject, or optionally, do not administer the treatment to the subject when the treatment is not selected for the subject.

10. A method of treating a subject having an eye disease, the method comprising: (a) Performing a first measurement on the subject to obtain at least two variables of a first set, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, eye anatomy, eye physiology, eye pathology, and changes thereof; (b) Calculating the at least two variables of the first set to produce a first composite biomarker; (c) Selecting a treatment for the subject based on the first composite biomarker; (d) Administering a first dose of the treatment to the subject; (e) Performing a second measurement on the subject to obtain the at least two variables of a second set; (f) Calculating the at least two variables of the second set to produce a second composite biomarker; (g) Determining whether the first dose of the treatment is therapeutically effective for treating the subject based on the first and second composite biomarkers; and (h) Administering a second dose of the treatment to the subject, wherein the second dose is different from the first dose when it is determined that the first dose is not therapeutically effective for treating the subject, and wherein the second dose is substantially the same as the first dose when it is determined that the first dose is therapeutically effective for treating the subject.

11. A method of treating a subject having an eye disease, the method comprising: (a) Performing a first measurement on the subject to obtain at least two variables of a first set, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, eye anatomy, eye physiology, eye pathology, and changes thereof; (b) Calculating the at least two variables of the first set to produce a first composite biomarker; (c) Administering a first dose of treatment to the subject; (d) Performing a second measurement on the subject to obtain the at least two variables of a second set; (e) Calculating the at least two variables of the second set to produce a second composite biomarker; (f) Determining that the first dose of the treatment is therapeutically effective for treating the subject when the difference between the first and second composite biomarkers exceeds a threshold, or optionally, determining that the first dose of the treatment is not therapeutically effective for treating the subject when the difference between the first and second composite biomarkers does not exceed the threshold; and (g) Administering a second dose of the treatment to the subject, wherein the second dose is different from the first dose when it is determined that the first dose is not therapeutically effective for treating the subject, and wherein the second dose is substantially the same as the first dose when it is determined that the first dose is therapeutically effective for treating the subject.

12. A method of monitoring the outcome of treatment for an ocular disease in a subject, the method comprising determining that a dose of the treatment administered to the subject is therapeutically effective for treating the subject by calculating at least two variables of the subject to produce a composite biomarker, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof.

13. A method of treating a subject having an ocular disease, which comprises administering to the subject a therapeutically effective dose of a treatment, wherein the treatment is selected for the subject based on a composite biomarker, wherein the composite biomarker is calculated based on at least two variables of the subject, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof.

14. A method, which comprises selecting a treatment for a subject having an ocular disease based on a composite biomarker, wherein the composite biomarker is calculated based on at least two variables of the subject, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular structure, ocular physiology, ocular pathology, and changes thereof.

15. A method of selecting a treatment for a subject having an ocular disease, the method comprising: (a) calculating at least two variables of the subject to produce a composite biomarker, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; and (b) selecting the treatment for the subject when the composite biomarker exceeds a threshold, or optionally, not selecting the treatment for the subject when the composite biomarker does not exceed the threshold.

16. A method of selecting a treatment for a subject having an ocular disease, the method comprising: (a) assaying the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) calculating the at least two variables to produce a composite biomarker; and (c) selecting the treatment for the subject when the composite biomarker exceeds a threshold, or optionally, not selecting the treatment for the subject when the composite biomarker does not exceed the threshold.

17. A method of predicting a subject's treatment response to a treatment for an ocular disease, the method comprising: (a) calculating at least two variables of the subject to produce a composite biomarker, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; and (b) predicting the subject's treatment response to the treatment based on the composite biomarker.

18. A method of predicting a subject's treatment response to a treatment for an ocular disease, the method comprising: (a) Measure the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) Calculate the at least two variables to generate a composite biomarker; and (c) When the composite biomarker exceeds a threshold, determine that the subject may have a therapeutic response to the treatment, or optionally, when the composite biomarker does not exceed the threshold, determine that the subject may not have a therapeutic response to the treatment.

19. A method comprising determining a dose of treatment for a subject with an ocular disease based on a composite biomarker, wherein the composite biomarker is calculated based on at least two variables of the subject, and wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof.

20. A method for determining a dose of treatment for a subject with an ocular disease, the method comprising: (a) Measure the subject to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) Calculate the at least two variables to generate a composite biomarker; and (c) Determine a dose of the treatment for the subject based on the composite biomarker.

21. A method for determining a dose of treatment for an ocular disease of a subject, the method comprising: (a) Perform a first measurement on the subject to obtain at least two variables of a first group, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof; (b) Calculate the at least two variables of the first group to generate a first composite biomarker; (c) Administer the treatment to the subject; (d) Perform a second measurement on the subject to obtain at least two variables of a second group; (e) Calculate the at least two variables of the second group to generate a second composite biomarker; and (f) Determine a dose of the treatment for the subject based on the first and second composite biomarkers.

22. A method comprising administering a second dose of treatment to a subject with an ocular disease, wherein based on a composite biomarker calculated from at least two variables of the subject, it is determined that a first dose of the treatment previously administered to the subject lacks therapeutic efficacy for treating the ocular disease of the subject, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and changes thereof, and wherein the second dose is different from the first dose.

23. A method for determining a dose of treatment for an ocular disease of a subject, the method comprising: (a) Administer a dose of the treatment to the subject; (b) Measure the object to obtain at least two variables, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and their changes; (c) Calculate the at least two variables to generate a composite biomarker; (d) Determine whether the dose of the treatment is therapeutically effective for treating the object suffering from the ocular disease based on the composite biomarker.

24. A method for determining a dose of treatment for an object suffering from an ocular disease, the method comprising: (a) Perform a first measurement on the object to obtain at least two variables of a first group, wherein at least one of the at least two variables is related to one or more selected from visual acuity, visual function, ocular anatomy, ocular physiology, ocular pathology, and their changes; (b) Calculate the at least two variables of the first group to generate a first composite biomarker; (c) Administer the dose of the treatment to the object; (d) Perform a second measurement on the object to obtain at least two variables of a second group; (e) Calculate the at least two variables of the second group to generate a second composite biomarker; and (f) When the difference between the first and second composite biomarkers exceeds a threshold, determine that the dose of the treatment is therapeutically effective for treating the object, or optionally, when the difference between the first and second composite biomarkers does not exceed the threshold, determine that the dose of the treatment is not therapeutically effective for treating the object.

25. The method according to any one of the preceding claims, wherein the ocular disease includes macular lesion, retinopathy, retinal atrophy, macular atrophy, macular degeneration, age-related macular degeneration (AMD), Stargardt disease (STGD), RP (retinitis pigmentosa), ABCA4 gene mutation, or a combination thereof.

26. The method according to any one of the preceding claims, wherein a sample from the object suffering from the ocular disease has an expression level of retinol-binding protein 4 (RBP4) of at least a threshold.

27. The method according to claim 26, wherein the threshold is about 25 μg / ml.

28. The method according to claim 26, wherein the threshold is about 35 μg / ml.

29. The method according to claim 26, wherein the threshold is between about 25 μg / ml and about 100 μg / ml.

30. The method according to any one of claims 26-29, wherein the expression level of RBP4 is measured by an assay comprising an antibody assay, an electrophoresis assay, an immunoassay, a radioimmunoassay, a chromatography assay, a mass spectrometry assay, a microarray-based detection assay, a polymerase chain reaction assay, a sequencing assay, an immunohistochemistry assay, or any combination thereof.

31. The method according to claim 30, wherein the antibody assay includes ELISA.

32. The method according to any one of claims 26 - 31, wherein a sample from the subject suffering from the eye disease has an expression level of vitamin A at least at a threshold value.

33. The method according to claim 32, wherein the threshold value is about 150 ng / mL.

34. The method according to claim 32, wherein the threshold value is about 225 ng / mL or about 390 ng / mL.

35. The method according to claim 32, wherein the threshold value is between about 150 ng / mL and about 500 ng / mL.

36. The method according to any one of claims 32 - 35, wherein the expression level of the vitamin is measured by an assay comprising an antibody assay, an electrophoresis assay, an immunoassay, a radioimmunoassay, a chromatography assay, a mass spectrometry assay, a microarray - based detection assay, a polymerase chain reaction assay, a sequencing assay, an immunohistochemistry assay, or any combination thereof.

37. The method according to claim 26 or 32, wherein the sample comprises a blood sample.

38. The method according to claim 37, wherein the expression level of RBP4 or vitamin A is measured from plasma or serum derived from the blood sample.

39. The method according to any one of claims 1 - 38, wherein the presence or absence of one or more genomic variants indicates that the subject suffers from the eye disease.

40. The method according to any one of claims 1 - 38, wherein the presence or absence of one or more genomic variants indicates that the subject suffers from Stargardt's disease.

41. The method according to any one of claims 1 - 40, wherein the presence or absence of one or more genomic variants selected from rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, and rs1801574 indicates that the subject suffers from the eye disease.

42. The method according to claim 39 or 40, wherein the one or more genomic variants comprise at least 4 of rs4147863, rs2275029, rs1800739, rs4147857, rs4147856, rs1801555, or rs1801574.

43. The method according to claim 39 or 40, wherein the one or more genomic variants comprise at least five of rs3747961, rs6666652, rs1800717, rs763108716, rs185601596, rs17110761, rs61748519, rs1801359, rs145766145, rs76258939, rs200551567, rs754765164, rs201602424, rs564661476, rs4147831, rs6657239, rs2297632, rs1801555, rs1762114, rs55860151, rs1800549, rs3112831, rs4147830, rs2297634 or rs4847281.

44. The method according to any one of claims 1-43, wherein the treatment comprises a pharmaceutical composition comprising an RBP4 inhibitor or a compound configured to reduce the blood RBP4 concentration of the subject.

45. The method according to claim 44, wherein the pharmaceutical composition comprises a compound having the structure of formula (I): Wherein: R A 1 、R A 2 、R A 3 、R A 4 and R A 5 each independently is H, halogen, CF 3 or C 1 -C 4 alkyl, wherein two or more of R A 1 、R A 2 、R A 3 、R A 4 and R A 5 are not H; R A 6 is H, OH or a halogen; and A A having a structure: Where α, β, χ and δ each independently do not exist or exist, and when present each Is a bond; X is C or N; Z 1 is N; Z 2 is N or NR A 9 , wherein R A 9 is H, C 1 -C 4 alkyl or oxetane; B A is a substituted or unsubstituted 5-, 6- or 7-membered ring structure; or a pharmaceutically acceptable salt thereof.

46. The method according to claim 45, wherein the compound has the structure or a pharmaceutically acceptable salt thereof.

47. The method according to claim 45, wherein the compound has the structure or a pharmaceutically acceptable salt thereof.

48. The method according to claim 44, wherein the pharmaceutical composition comprises a compound having the structure of formula (II): Wherein: Ring A B is an optionally further substituted benzene; R B 1 is an optionally substituted branched C 3 -C 6 alkyl group; X B 1 is O, S, SO, SO 2 or NH; X B 2 is a key or C 1 -C 3 an alkylene group; Ring B B is azetidine or piperidine; X B 3 is CO or SO 2 ; R B 2 is an optionally substituted hydrocarbyl group, an optionally substituted heterocyclic group, an optionally substituted hydroxy group, an optionally substituted mercapto group, a cyano group, a nitro group, an acyl group or a halogen atom; or Its pharmaceutically acceptable salts.

49. The method according to claim 48, wherein the compound is 4-(3-(2-tert-butylphenoxy)azetidin-1-yl)-4-oxobutyric acid, 3-{3-[(2-tert-butyl-4-fluorophenoxy)methyl]azetidin-1-yl}-3-oxopropanoic acid, 2-{[3-(2-tert-butyl-4-chlorophenoxy) Azetidin-1-yl]carbonyl}pyridine, 4-[3-(2-tert-butyl-4-chlorophenoxy)azetidin-1-yl]-4-oxobutyric acid, {3-[(2-tert-butyl-4-chlorophenoxy)methyl]azetidin-1-yl}(oxo)acetic acid, {3-[(2-tert-butylphenoxy)methyl]azetidin-1-yl}(oxo)acetic acid, 3-{3-[(2-tert-butylphenoxy)methyl]azetidin-1-yl}-3-oxopropanoic acid, {4-[(2-tert-butyl-4-chlorophenoxy)methyl]piperidin-1-yl}(oxo)acetic acid, [4-(2-tert-butylphenoxy)piperidin-1-yl](oxo)acetic acid or {4-[(2-tert-butylphenoxy)methyl]piperidin-1-yl}(oxo)acetic acid, or its pharmaceutically acceptable salts.

50. The method according to claim 44, wherein the pharmaceutical composition comprises a compound having the structure of formula (III): Wherein: Ring A C is a benzene ring optionally substituted with 1 to 3 substituents selected from the following: (a) a halogen atom and (b) C 1 -C 6 alkyl; Ring B C is a piperazine ring optionally substituted with 1 to 3 substituents selected from the following: (a) a halogen atom, (b) C 1 -C 6 alkyl optionally substituted with 1 to 3 halogen atoms, and (c) C 1 -C 6 alkoxy; and R C is (1) optionally substituted C 1 -C 10 alkyl, (2) optionally substituted C 6 -C 14 aryl, (3) optionally substituted 5- or 6-membered aromatic heterocyclic group, (4) optionally substituted amino group, (5) optionally substituted carboxyl group or (6) optionally substituted carbamoyl group; or Its pharmaceutically acceptable salts.

51. The method according to claim 50, wherein the compound is N-{[4-(2-tert-butylphenyl)piperazin-1-yl]carbonyl}glycine, 3-[4-(2-tert-butylphenyl)piperazin-1-yl]-3-oxopropanoic acid, [4-(2-tert-butyl-4-chlorophenyl)piperazin-1-yl](oxo)acetic acid, 5-{2-[4-(2-tert-butylphenyl)piperazin-1-yl]-2-oxoethyl}imidazolidine-2,4-dione, [(5-{[4-(2-tert-butylphenyl)piperazin-1-yl]carbonyl}isoxazol-3-yl)oxy]acetic acid, or a pharmaceutically acceptable salt thereof.

52. The method according to claim 44, wherein the pharmaceutical composition comprises a compound having the structure of formula (IV): Wherein: Ring A D is a 5-membered non-aromatic heterocycle, which is optionally further substituted by an oxo group; Ring B D is a benzene ring, which is optionally further substituted with 1 to 4 substituents; and X D is O, CH 2 O, OCH 2 , CH 2 , (CH 2 ) 2 , S, CH 2 S, SCH 2 , S(O), CH 2 S(O), S(O)CH 2 , S(O) 2 , CH 2 S(O) 2 or S(O) 2 CH 2 ; or a pharmaceutically acceptable salt thereof.

53. The method according to claim 52, wherein the compound is ((3S)-1-[3,5-bis(trifluoromethyl)phenyl]pyrrolidin-3-yl)oxyacetic acid, ((1-[4-chloro-3-(trifluoromethyl)phenyl]pyrrolidin-3-yl)thio)acetic acid, 3-{(2R,5S)-5-[3,5-bis(trifluoromethyl)phenyl]tetrahydrofuran-2-yl}propanoic acid, or a pharmaceutically acceptable salt thereof.

54. The method according to claim 44, wherein the pharmaceutical composition comprises a compound having the structure of formula (V): Wherein: Ring A E is a pyrazole ring, pyridine ring, oxazole ring, imidazole ring or pyrimidine ring; X E is S, an optionally substituted alkylene or O; and R E is a hydrogen atom or a C 1 -C 6 -alkyl; or its pharmaceutically acceptable salt.

55. The method according to claim 54, wherein the compound is ((4-(3,5-bis(trifluoromethyl)phenyl)-1,3-oxazol-2-yl)thio)acetic acid, ethyl ((6-(3,5-bis(trifluoromethyl)phenyl)pyridin-3-yl)thio)acetic acid, ((6-(3,5-bis(trifluoromethyl)phenyl)pyridin-3-yl)thio)acetic acid or 3-(3-(3,5-bis(trifluoromethyl)phenyl)-1H-pyrazol-1-yl)butyric acid or 3-{3-[3,5-bis(trifluoromethyl)phenyl]-1H-pyrazol-1-yl}propanoic acid (also known as STG-001).

56. The method according to any one of claims 1-55, wherein at least one of the at least two variables is correlated with one or more selected from visual acuity, the average score of a functional vision questionnaire, and measurements obtained from imaging techniques.

57. The method according to any one of claims 1-55, wherein at least one of the at least two variables is correlated with one or more selected from a change in visual acuity, a change in the average score of a functional vision questionnaire, and a change in measurements obtained from imaging techniques for detecting or quantifying changes in ocular structure or visual function.

58. The method according to any one of claims 1-55, wherein at least one of the at least two variables is correlated with a change in visual acuity, a change in the average score of a functional vision questionnaire, and a change in measurements obtained from imaging techniques for detecting or quantifying optic nerve atrophy or changes in visual function.

59. The method according to any one of claims 1-55, wherein the at least two variables are related to a change in visual acuity, a change in the average score of a functional vision questionnaire, and a change in a measurement obtained from an imaging technique for detecting or quantifying a change in optic nerve atrophy or visual function.

60. The method according to any one of claims 1-55, wherein the at least two variables are related to a change in visual acuity and a change in the average score of a functional vision questionnaire.

61. The method according to any one of claims 1-55, wherein the at least two variables are related to a change in visual acuity and a change in a measurement obtained from an imaging technique for detecting or quantifying a change in optic nerve atrophy or visual function.

62. The method according to any one of claims 1-55, wherein the at least two variables are related to a change in the average score of a functional vision questionnaire and a change in a measurement obtained from an imaging technique for detecting or quantifying a change in optic nerve atrophy or visual function.

63. The method according to any one of claims 1-55, wherein both of the at least two variables are related to a change in visual acuity.

64. The method according to any one of claims 1-55, wherein both of the at least two variables are related to a change in the average score of a functional vision questionnaire.

65. The method according to any one of claims 1-55, wherein both of the at least two variables are related to a change in a measurement obtained from an imaging technique for detecting or quantifying a change in optic nerve atrophy or visual function.

66. The method according to any one of claims 1-55, wherein at least one of the at least two variables is related to a change in an eye structure, a change in an eye pathology, a change in retinal atrophy, a change in macular atrophy, a change in macular degeneration, or a combination thereof.

67. The method according to any one of claims 1-66, wherein two of the at least two variables are related to each other.

68. The method according to any one of claims 1-66, wherein one of the at least two variables is related to visual acuity.

69. The method according to any one of claims 1-66, wherein one of the at least two variables is related to a change in visual acuity.

70. The method according to any one of claims 1-66, wherein all of the at least two variables are related to a change in visual acuity.

71. The method according to any one of claims 1-66, wherein one of the at least two variables is related to the state of the eye disease.

72. The method according to any one of claims 1-66, wherein one of the at least two variables is related to the progression of the eye disease.

73. The method according to any one of claims 1-66, wherein the at least two variables include values obtained from a logMAR chart, a Snellen chart, a best corrected visual acuity (BCVA) test, an Early Treatment Diabetic Retinopathy Study (ETDRS) letter test, or a combination thereof.

74. The method according to any one of claims 1 - 66, wherein the at least two variables include values obtained from a functional vision questionnaire, which preferably includes one or more questions related to the following: (a) The confidence of the subject in moving to different places during the day or at night; (b) The ability of the subject to recognize people, participate in sports, or obtain information; and (c) The ability of the subject to engage in social interaction or work efficiency, wherein each of the one or more questions is scored for the subject.

75. The method according to claim 74, wherein each of the one or more questions is scored for the subject on a severity scale of 0 - 4 as follows: (a) Scored 0 for an answer of "never"; (b) Scored 1 for an answer of "almost never"; (c) Scored 2 for an answer of "sometimes"; (d) Scored 3 for an answer of "almost always"; and (e) Scored 4 for an answer of "always".

76. The method according to any one of claims 1 - 75, wherein the at least two variables include values obtained from microperimetry (MP), perimetry, or both.

77. The method according to claim 76, wherein the MP includes a spatial map measuring the retinal sensitivity of the subject.

78. The method according to claim 76, wherein the MP includes measuring the average retinal sensitivity of the subject or its change.

79. The method according to any one of claims 1 - 78, wherein the at least two variables include values obtained from one or more selected from fundus photography, fundus angiography (FA), fundus autofluorescence (FAF), spectral domain - optical coherence tomography (SD - OCT), quantitative autofluorescence (qAF), infrared (IR) imaging, optical coherence tomography - angiography (OCT - A), and wide - field imaging.

80. The method according to any one of claims 1 - 79, wherein the at least two variables include one or more selected from the area of suspected decreased autofluorescence (QDAF), the area of determined decreased autofluorescence (DDAF), the area of decreased autofluorescence (DAF), retinal thickness, ellipsoid zone (EZ) defect width, central subfield retinal thickness (CST), outer subfield retinal thickness (OST), middle subfield retinal thickness (MST), and their changes.

81. The method according to claim 80, wherein the area of decreased autofluorescence (DAF) includes the area of suspected decreased autofluorescence (QDAF) and the area of determined decreased autofluorescence (DDAF).

82. The method according to any one of claims 1 - 81, wherein the at least two variables include two or more selected from visual acuity, QDAF, DAF, EZ defect width, or their changes.

83. The method according to any one of claims 1 - 81, wherein the at least two variables include visual acuity, QDAF, DAF, EZ defect width, or their changes.

84. The method according to claim 83, wherein the EZ defect width is related to the visual acuity.

85. The method according to claim 83, wherein the EZ defect width is related to the QDAF.

86. The method according to claim 83, wherein the EZ defect width is related to the DAF.

87. The method according to claim 83, wherein the visual acuity is related to the QDAF.

88. The method according to claim 83, wherein the EZ defect width is related to the visual acuity, QDAF, and DAF.

89. The method according to claim 83, wherein the visual acuity is related to the EZ defect width and QDAF.

90. The method according to claim 83, wherein the EZ defect width is related to the visual acuity, QDAF, and DAF, and the correlation coefficient is greater than about 0.8 and the p-value is less than about 0.

05.

91. The method according to claim 83, wherein the visual acuity is related to the EZ defect width and QDAF, and the p-value is less than about 0.

05.

92. The method according to any one of claims 1-91, wherein the at least two variables of the subject are obtained from the right eye and the left eye of the subject.

93. The method according to any one of claims 1-92, wherein the method further comprises predicting whether the prognostic potential of the treatment is improved based on one or more composite biomarkers.

94. The method according to any one of claims 1-93, wherein when the composite biomarker exceeds the threshold, the treatment is therapeutically effective for treating the subject.

95. The method according to any one of claims 1-93, wherein when the composite biomarker does not exceed the threshold, the treatment is not therapeutically effective for treating the subject.

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