LRRC4 family mimic molecules and their diagnostic applications

Abstract

The present invention relates to the use of LRRC4 family mimetic molecules to identify expression levels of FAM19A5 protein in a subject. In some aspects, such mimetic molecules can be used to identify and / or diagnose subjects suffering from or at risk of developing a disease or disorder associated with increased FAM19A5 protein expression. The present invention also provides methods of treating subjects identified and / or diagnosed using the methods provided herein.

Description

Detailed Description of the Invention

[0001] [Technical field] CROSS-REFERENCE TO RELATED APPLICATIONS This PCT application claims priority to U.S. Provisional Application No. 63 / 282,598, filed November 23, 2021, the entire contents of which are incorporated herein by reference.

[0002] References to sequence listings submitted electronically using EFS-Web The entire sequence content, submitted electronically as an XML file (Name: 3763.020PC01_Seqlisting_ST26, Size: 130,483 bytes; Creation Date: November 22, 2022), is incorporated by reference into this application.

[0003] The present invention provides a method for measuring the expression level of FAM19A5 protein using LRRC4 family mimetic molecules. The present invention also provides the use of said LRRC4 family mimetic molecules to diagnose a disease or disorder associated with increased FAM19A5 expression and / or to identify subjects suitable for treatment with a FAM19A5 antagonist.

[0004] [Background technology] Diseases and disorders of the central nervous system (CNS) comprise a heterogeneous group of diseases with etiology and pathogenesis that are generally unknown. Currently, there is no way to identify subjects who will respond to treatment for CNS diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), and other related disorders such as traumatic brain injury (TBI), neuropathic pain, and glaucoma, and there are no treatments for them. In fact, in the vast majority of cases, available treatments for CNS diseases only provide some minor symptomatic benefit and maintain a palliative state. With increasing life expectancy and population growth worldwide, it is expected that more individuals will suffer from CNS diseases and disorders (see Feigin, VL, et al, Lancet Neurol 16(11):877-897(2017)). Thus, there is a need for new and more effective approaches to treating and / or diagnosing CNS disorders.

[0005] Summary of the Invention [Means for solving the problems] The present invention provides a method for measuring the expression level of family with sequence similarity 19, member A5 ("FAM19A5") protein in a subject in need thereof, the method comprising the steps of (i) contacting a biological sample from the subject with a leucine-rich containing 4 ("LRRC4") family mimetic molecule capable of specifically binding to the FAM19A5 protein, and (ii) measuring the expression level of the FAM19A5 protein ("FAM19A5 protein expression level") from the biological sample.

[0006] The present invention also provides a method for diagnosing a disease or disorder in a subject in need thereof, the method comprising the steps of (i) contacting a biological sample from the subject with a leucine-rich containing 4 ("LRRC4") family mimetic molecule capable of specifically binding to family with sequence similarity 19, member A5 ("FAM19A5") protein, and (ii) measuring an expression level of the FAM19A5 protein ("FAM19A5 protein expression level") from the biological sample.

[0007] In some aspects, the disease or disorder is associated with an increased FAM19A5 protein expression level. In some aspects, the FAM19A5 protein expression level is increased in the biological sample compared to a control group (e.g., a corresponding expression level in a subject not afflicted with the disease or disorder). In some aspects, the FAM19A5 protein expression is increased about 1-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 6-fold, about 7-fold, about 8-fold, about 9-fold, about 10-fold, about 15-fold, about 20-fold, about 25-fold, about 30-fold, about 35-fold, about 40-fold, about 45-fold, or about 50-fold compared to a control group.

[0008] The invention also provides a method for identifying a subject suitable for treatment with an antagonist to Family with sequence similarity 19, member A5 ("FAM19A5") protein (a "FAM19A5 antagonist"), the method comprising the steps of (i) contacting a biological sample from the subject with a leucine-rich containing 4 ("LRRC4") family mimetic molecule capable of specifically binding to FAM19A5 protein, and (ii) measuring an expression level of FAM19A5 protein (a "FAM19A5 protein expression level") from the biological sample, the subject being suitable for treatment if the FAM19A5 protein expression level is increased compared to a control group (e.g., a corresponding expression level in a subject not afflicted with the disease or disorder). In some aspects, the subject has previously received a FAM19A5 antagonist.

[0009] In some aspects, the FAM19A5 protein expression level is reduced by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% compared to a control group.

[0010] In some aspects, the methods of identifying a subject suitable for FAM19A5 antagonist treatment provided herein further comprise administering to the subject an additional dose of a FAM19A5 antagonist after the contacting and measuring steps.

[0011] The present invention also provides a method for treating a disease or disorder associated with increased expression levels of Family with sequence similarity 19, member A5 ("FAM19A5") protein in a subject in need thereof, the method comprising the steps of administering to the subject a therapeutic agent for the disease or disorder, and measuring the expression level of FAM19A5 protein ("FAM19A5 protein expression level") by contacting a biological sample from the subject with a leucine-rich containing 4 ("LRRC4") family mimetic molecule capable of specifically binding to FAM19A5 protein.

[0012] In some aspects, the methods of treatment provided herein further comprise administering to the subject an additional dose of a therapeutic agent if the FAM19A5 protein expression level is increased compared to a control group (e.g., a corresponding expression level in a subject not afflicted with a disease or disorder). In some aspects, the FAM19A5 protein expression level is increased by about 1-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 6-fold, about 7-fold, about 8-fold, about 9-fold, about 10-fold, about 15-fold, about 20-fold, about 25-fold, about 30-fold, about 35-fold, about 40-fold, about 45-fold, or about 50-fold compared to a control group.

[0013] For any of the methods provided herein, in some aspects, the FAM19A5 protein expression level is measured by enzyme linked immunosorbent assay (ELISA), immunohistochemistry, Western-blotting, radioimmunoassay, radical-immunodiffusion, immunoprecipitation assay, Ouchterlony immunodiffusion method, rocket immunoelectrophoresis, tissue immunostaining method, complement fixation assay, FACS, protein chip, SIMOA technology, or a combination thereof. In some aspects, the biological sample comprises blood, cerebral spinal fluid (CSF), serum, plasma, tissue, cell culture medium, saliva, urine, or a combination thereof. In some aspects, the contacting and measuring steps are performed outside the body.

[0014] For any of the methods of treatment provided herein, in some aspects the treatment includes an antagonist to FAM19A5 ("FAM19A5 antagonist"), a leucine-rich containing 4 ("LRRC4") family mimetic molecule, or both.

[0015] In some aspects, the FAM19A5 antagonist comprises an antibody or an antigen-binding portion thereof, an antisense oligonucleotide, an siRNA, an shRNA, an miRNA, a dsRNA targeting FAM19A5, an aptamer, a PNA, a vector comprising the same, or a combination thereof. In some aspects, the FAM19A5 antagonist is an antibody or an antigen-binding portion thereof.

[0016] In some aspects, the LRRC4 family mimetic molecule of the methods provided herein is not an antibody or an antigen-binding portion thereof.

[0017] In some aspects, the LRRC4 family mimetic molecule comprises a polypeptide that comprises, consists of, or consists essentially of a domain of a LRRC4 protein family member, wherein the domain (the "FAM19A5 binding domain") is capable of binding to a FAM19A5 protein, and wherein the polypeptide is shorter than a corresponding full-length LRRC4 protein family member (SEQ ID NO:4; SEQ ID NO:5; or SEQ ID NO:6).

[0018] In some aspects, the FAM19A5 binding domain of the LRRC4 family mimetic molecule is about 10 to about 23 amino acids in length. In some aspects, the FAM19A5 binding domain is about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, or about 23 amino acids in length. In some aspects, the FAM19A5 binding domain is about 10 amino acids in length.

[0019] For any of the methods provided herein, in some aspects, the LRRC4 family mimetic molecule comprises a polypeptide comprising an amino acid sequence (N-terminus to C-terminus) having the following chemical formula: A-(T / S)-B (Formula I): where (i) A is X1-(T / S)-(Y / F)-F-X5; X1 is tyrosine (Y), phenylalanine (F), valine (V), leucine (L), or isoleucine (I); (T / S) is threonine (T) or serine (S); (Y / F) is tyrosine (Y) or phenylalanine (F); X5 is any amino acid; (ii) B comprises (V / I)-TV-(E / V); (V / I) is valine (V) or isoleucine (I); (E / V) is glutamic acid (E) or valine (V).

[0020] For any of the methods provided herein, in some aspects, the LRRC4 family mimetic molecule comprises a polypeptide comprising an amino acid sequence (N-terminus to C-terminus) having the following formula: A-(T / S)-B (Formula I): where (i) A comprises (Y / W / M)-(T / Y)-(Y / W)-(F / Y / W)-(T / Y); (Y / W / M) is tyrosine (Y), tryptophan (W), or methionine (M); (T / Y) is threonine (T) or tyrosine (Y); (Y / W) is tyrosine (Y) or tryptophan (W); (F / Y / W) is phenylalanine (F), tyrosine (Y), or tryptophan (W); (ii) B includes X7-(T / S / Y)-X9-X10; X7 is valine (V), tyrosine (Y), phenylalanine (F), leucine (L), tryptophan (W), or methionine (M); (T / S / Y) is threonine (T), serine (S), or tyrosine (Y); X9 is valine (V), isoleucine (I), tyrosine (Y), phenylalanine (F), leucine (L), tryptophan (W), or methionine (M); X10 is glutamic acid (E), aspartic acid (D), isoleucine (I), tyrosine (Y), phenylalanine (F), methionine (M), or tryptophan (W).

[0021] For any of the methods provided herein, in some aspects, the LRRC4 family mimetic molecule comprises an amino acid sequence (N-terminus to C-terminus) having the following formula: X1-X2-X3-F-X5-T-X7-TV-X10 (Formula II): where X1 is Y, F, V, L, or I; X2 is T or S; X3 is Y or F; X5 is any amino acid; X7 is V or I; and / or X10 is E or V.

[0022] For any of the methods provided herein, in some aspects, the LRRC4 family mimetic molecule comprises a polypeptide comprising an amino acid sequence (N-terminus to C-terminus) having the following formula: X1-X2-X3-X4-X5-X6-X7-X8-X9-X10 (Formula VI): where X1 is Y, F, V, L, I, W, or M; X2 is T, S, or Y; X3 is Y, F, or W; X4 is F, Y, or W; X5 is any amino acid, e.g., T, S, or Y; X6 is T, S, or Y; X7 is V, I, Y, F, L, W, or M; X8 is T, S, or Y; X9 is V, I, Y, F, L, W, or M; and / or X10 is E, D, V, I, Y, F, M, or W.

[0023] In some aspects, X1 is Y, F, V, L, or I; X2 is T or S; X3 is Y or F; X4 is F; X5 is T or S; X6 is T; X7 is V or I; X8 is T; X9 is V; X10 is E or V; or a combination thereof.

[0024] In some aspects, the LRRC4 family mimetic molecule comprises a polypeptide comprising the amino acid sequence provided as SEQ ID NO:29 (YTYFTTVTVE). In some aspects, the LRRC4 family mimetic molecule comprises a polypeptide consisting of the amino acid sequence provided as SEQ ID NO:29 (YTYFTTVTVE). In some aspects, the LRRC4 family mimetic molecule comprises a polypeptide comprising the amino acid sequence provided as SEQ ID NO:20 (NYSFFTTVTVETTEISPEDTTRK). In some aspects, the LRRC4 family mimetic molecule comprises a polypeptide consisting of the amino acid sequence provided as SEQ ID NO:20 (NYSFFTTVTVETTEISPEDTTRK). In some aspects, the polypeptide comprises a polypeptide comprising the amino acid sequence provided as SEQ ID NO:21 (NFSYFSTVTVETMEPSQDERTTR). In some aspects, the polypeptide comprises a polypeptide consisting of the amino acid sequence provided as SEQ ID NO:21 (NFSYFSTVTVETMEPSQDERTTR). In some aspects, the LRRC4 family mimetic molecule comprises a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 18 (GYTYFTTVTVETLETQPGEE). In some aspects, the LRRC4 family mimetic molecule comprises a polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 18 (GYTYFTTVTVETLETQPGEE).

[0025] In some aspects, the amino acid at X2 is phosphorylated or O-glycosylated.

[0026] In some aspects, the LRRC4 family mimetic molecule comprises a polypeptide comprising an amino acid sequence having at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:29 (YTYFTTVTVE).

[0027] In some aspects, the LRRC4 family mimetic molecule comprises a polypeptide comprising an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence set forth in SEQ ID NO:5, SEQ ID NO:4, or SEQ ID NO:6, respectively, and that comprises at least one amino acid sequence modification relative to the amino acid sequence set forth in SEQ ID NO:5, SEQ ID NO:4, or SEQ ID NO:6. In some aspects, the at least one amino acid modification increases binding of the polypeptide to a FAM19A5 protein. In some aspects, the at least one amino acid modification increases stability of the LRRC4 family mimetic molecule.

[0028] In some aspects, at position 453 of SEQ ID NO:5, the amino acid residue is T or modified with S or Y; at position 454 of SEQ ID NO:5, the amino acid residue is T or modified with S or Y; at position 449 of SEQ ID NO:5, the amino acid residue is Y or modified with F, V, L, I, W, or M; at position 450 of SEQ ID NO:5, the amino acid residue is T or modified with S or Y; at position 451 of SEQ ID NO:5, the amino acid residue is Y or modified with F or W; at position 452 of SEQ ID NO:5 at position 455 of SEQ ID NO:5, the amino acid residue is V or modified with I, Y, F, L, W, or M; at position 456 of SEQ ID NO:5, the amino acid residue is T or modified with S or Y; at position 457 of SEQ ID NO:5, the amino acid residue is V or modified with I, Y, F, L, W, or M; at position 458 of SEQ ID NO:5, the amino acid residue is E or modified with D, V, I, Y, F, M, or W; or combinations thereof.

[0029] In some aspects, the LRRC4 family mimetic molecule does not include the transmembrane and / or intracellular domain of a LRRC4 protein family member.

[0030] In some aspects, the LRRC4 family mimetic molecule further comprises one or more additional amino acids at the N-terminus, C-terminus, or both the N-terminus and C-terminus of the LRRC4 family mimetic molecule, in some aspects, the one or more additional amino acids are hydrophilic amino acids, D-amino acids, or both.

[0031] In some aspects, the N-terminus, C-terminus, or both the N-terminus and C-terminus of the LRRC4 family mimetic molecule comprise a modification that increases the stability of the LRRC4 family mimetic molecule. In some aspects, the modification comprises Fmoc, PEGylation, acetylation, methylation, cyclization, or a combination thereof.

[0032] In some aspects, the LRRC4 family mimetic molecule is a fusion protein. In some aspects, the LRRC4 family mimetic molecule further comprises a half-life extending moiety. In some aspects, the half-life extending moiety comprises Fc, albumin, albumin binding polypeptide, Pro / Ala / Ser (PAS), C-terminal peptide of the beta subunit of human chorionic gonadotropin (CTP), polyethylene glycol (PEG), long unstructured hydrophilic sequences of amino acids (XTEN), hydroxyethyl starch (HES), an albumin binding small molecule, or a combination thereof.

[0033] In some aspects, the LRRC4 family mimetic molecule comprises a small molecule.

[0034] In some aspects, the LRRC4 family mimetic molecule is a small molecule of formula (I):

[0035] [ka]

[0036] or a pharma- ceutically acceptable salt thereof: In the above, (i) R1, R2, and R3 are each independently selected from the group consisting of hydrogen, fluoro, chloro, bromo, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, t-butyl, n-pentyl, iso-pentyl, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 1-fluoroethyl, 2,2-difluoroethyl, 1,2-difluoroethyl, 1,1-difluoroethyl, and the like. roethyl, 2,2,2-trifluoroethyl, methoxy, ethoxy, n-propyloxy, iso-propyloxy, n-butoxy, trifluoromethoxy, difluoromethoxy, fluoromethoxy, acetyl, propionyl, n-butanoyl, iso-butanoyl, n-pentanoyl, nitro, amino, N-methylamino, N-ethylamino, Nn-propylamino, N,N-dimethylamino, N,independently selected from N-dimethylamino, N-acetylamino, N-propionylamino, N-(trifluoroacetyl)amino, formyl, hydroxy, methylthio, ethylthio, n-propylthio, methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, phenyl, hydroxymmethyl, 1-hydroxyethyl, and 2-hydroxyethyl; (ii)

[0037] [ka]

[0038] is a single or double bond; (iii) Z is a linear or branched (C1-C8) alkyl, a linear or branched (C2-C8) alkenyl, a linear or branched (C2-C8) alkynyl, a (C3-C8) cycloalkyl, a (C5-C8) cycloalkenyl, a (3-8 membered) heterocycloalkyl, a (C7-C 14 )bicycloalkyl, (C7-C 14 )bicycloalkenyl, (7-14 membered)heterobicycloalkyl, (C6-C 10 )aryl, (5-10 membered)heteroaryl, and -CH-C(O)-CH=CH-Q, where Q is selected from (C3-C8)cycloalkyl, (C5-C8)cycloalkenyl, (3-8 membered)heterocycloalkyl, (C6-C 10)aryl, and (5-6 membered)heteroaryl; each cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl is optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from (C1-C6)alkoxy, (C1-C6)alkyl, halo, (C1-C6)haloalkoxy, nitro, amino, N-methylamino, N-ethylamino, N-N-propylamino, N,N-dimethylamino, formyl, and hydroxy; (iv) L is a single, double, or triple bond.

[0039] In some aspects, the LRRC4 family mimetic molecule is a mimetic molecule selected from the following structural formulas:

[0040] [ka]

[0041] or a pharma- ceutically acceptable salt thereof.

[0042] In some aspects, the LRRC4 family mimetic molecule is a small molecule of formula (II):

[0043] [ka]

[0044] or a pharma- ceutically acceptable salt thereof: where (i) R1, R2 and R3 are hydrogen, fluoro, chloro, bromo, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, t-butyl, n-pentyl, iso-pentyl, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 1-fluoroethyl, 2,2-difluoroethyl, 1,2-difluoroethyl, 1,1-difluoroethyl, 2,2,2-trifluoroethyl, methoxy, ethoxy, n-furopyroxy, iso-furopyroxy, n-butoxy, trifluoromethoxy, difluoromethyl, independently selected from methoxy, fluoromethoxy, acetyl, propionyl, n-butanoyl, iso-butanoyl, n-pentanoyl, nitro, amino, N-methylamino, N-ethylamino, Nn-propylamino, N,N-dimethylamino, N-acetylamino, N-propionylamino, N-(trifluoroacetyl)amino, formyl, hydroxy, methylthio, ethylthio, n-propylthio, methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, phenyl, hydroxymethyl, 1-hydroxyethyl, and 2-hydroxyethyl; (ii) Z is a linear or branched (C1-C8) alkyl, a linear or branched (C2-C8) alkenyl, a linear or branched (C2-C8) alkynyl, a (C3-C8) cycloalkyl, a (C5-C8) cycloalkenyl, a (3-8 membered) heterocycloalkyl, a (C7-C 14 )bicycloalkyl, (C7-C 14 )bicycloalkenyl, (7-14 membered)heterobicycloalkyl, (C6-C 10 )aryl, (5-10 membered)heteroaryl, and -CH=CH-Q, where Q is selected from (C3-C8)cycloalkyl, (C5-C8)cycloalkenyl, (3-8 membered)heterocycloalkyl, (C6-C 10)aryl, and (5-6 membered)heteroaryl; each cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl is optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from (C1-C6)alkoxy, (C1-C6)alkyl, halo, (C1-C6)haloalkoxy, nitro, amino, N-methylamino, N-ethylamino, N-N-propylamino, N,N-dimethylamino, formyl, and hydroxy; (iii) L is a single, double, or triple bond.

[0045] In some aspects, the LRRC4 family mimetic molecule is a mimetic molecule selected from the following structural formulas:

[0046] [ka]

[0047] or a pharma- ceutically acceptable salt thereof.

[0048] In some aspects, the LRRC4 family mimetic molecule is a small molecule of formula (III):

[0049] [ka]

[0050] or a pharma- ceutically acceptable salt thereof: where (i) R1, R2 and R3 are hydrogen, fluoro, chloro, bromo, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, t-butyl, n-pentyl, iso-pentyl, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 1-fluoroethyl, 2,2-difluoroethyl, 1,2-difluoroethyl, 1,1-difluoroethyl, 2,2,2-trifluoroethyl, methoxy, ethoxy, n-furopyroxy, iso-furopyroxy, n-butoxy, trifluoromethoxy, difluoromethyl, independently selected from methoxy, fluoromethoxy, acetyl, propionyl, n-butanoyl, iso-butanoyl, n-pentanoyl, nitro, amino, N-methylamino, N-ethylamino, Nn-propylamino, N,N-dimethylamino, N-acetylamino, N-propionylamino, N-(trifluoroacetyl)amino, formyl, hydroxy, methylthio, ethylthio, n-propylthio, methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, phenyl, hydroxymethyl, 1-hydroxyethyl, and 2-hydroxyethyl; (ii) Z is a linear or branched (C1-C8) alkyl, a linear or branched (C2-C8) alkenyl, a linear or branched (C2-C8) alkynyl, -Y-(C3-C8) cycloalkyl, -Y-(C5-C8) cycloalkenyl, -Y-(3-8 membered) heterocycloalkyl, -Y-(C7-C 14 )bicycloalkyl, -Y-(C7-C 14 )bicycloalkenyl, -Y-(7-14 membered)heterobicycloalkyl, -Y-(C6-C 10 )aryl, and -Y-(5-10 membered)heteroaryl, wherein Y is a bond or a C1-C3 straight or branched alkylene, and said cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from (C1-C6)alkoxy, (C1-C6)alkyl, halo, (C1-C6)haloalkoxy, nitro, amino, N-methylamino, N-ethylamino, N-N-propylamino, N,N-dimethylamino, formyl, and hydroxy; (iii) L is a single, double, or triple bond; (iv) n is 0 or 1.

[0051] In some aspects, the LRRC4 family mimetic molecule is a mimetic molecule selected from the following structural formulas:

[0052] [ka]

[0053] or a pharma- ceutically acceptable salt thereof.

[0054] In some aspects, the LRRC4 family mimetic molecule may compete with a LRRC4 protein family member for binding to the FAM19A5 protein. In some aspects, the LRRC4 protein family member comprises a LRRC4 protein, a LRRC4B protein, a LRRC4C protein, or a combination thereof.

[0055] In some aspects, the disease or disorder comprises amyotrophic lateral sclerosis (ALS), Alzheimer's disease, glaucoma, diabetic retinopathy, neuropathic pain, spinal cord injury, traumatic brain injury, stroke, Parkinson's disease, or a combination thereof.

[0056] BRIEF DESCRIPTION OF THE DRAWINGS 1A-1C provide schematic diagrams of different example approaches using a sandwich ELISA assay to measure FAM19A5 protein expression from a sample. FIG. 1A shows the use of two different anti-FAM19A5 antibodies (Antibody A conjugated to HRP and Antibody B coated on an ELISA plate) in a sandwich ELISA assay to measure FAM19A5 protein expression from a sample. FIG. 1B shows the use of an LRRC4 family protein (LRRC4B) conjugated to Fc ("LRRC4B-hFc") and an anti-FAM19A5 antibody (Antibody A conjugated to HRP) in a sandwich ELISA assay to measure FAM19A5 protein expression from a sample. FIG. 1C shows the use of a LRRC4 family protein (e.g., LRRC4B) bound to a 6xHis tag, a Tobacco Etch Virus (TEV) protease sequence ("His-TEV-LRRC4B"), and an anti-FAM19A5 antibody (Antibody A conjugated to HRP) in a sandwich ELISA assay to measure FAM19A5 protein expression from samples.

[0057] FIG. 2 provides a comparison of FAM19A5 protein levels in the blood of mice (wild type and FAM19A knockout), rats, monkeys (cynomolgus and marmosets) and humans, as measured by the LRRC4 family mimetic molecule-based sandwich ELISA assay described in FIG.

[0058] Figure 3 shows FAM19A5 protein levels in rats after treatment with anti-FAM19A5 antibody. Rats received a single intravenous dose of anti-FAM19A5 antibody at one of two doses, 10 mg / kg ("1") or 50 mg / kg ("2"). Blood was collected from the animals at various times after administration, and FAM19A5 protein levels were measured using the LRRC4 family mimetic molecule-based sandwich ELISA described in Figure 1.

[0059] Figure 4 shows FAM19A5 protein and anti-FAM19A5 antibody levels in blood and CSF, respectively, of rats after anti-FAM19A5 antibody treatment. Rats received a single intravenous dose of anti-FAM19A5 antibody (10 mg / kg). "Basal" = level before anti-FAM19A5 antibody administration. FAM19A5 protein levels were measured using the LRRC4 family mimetic molecule-based sandwich ELISA assay described in Figure 1.

[0060] Figure 5 shows FAM19A5 protein and anti-FAM19A5 antibody levels in blood of rats administered anti-FAM19A5 antibody intravenously every week. Rats received a total of four doses (10 mg / kg per dose). Anti-FAM19A5 antibody levels were measured using an ELISA method using rabbit anti-human IgG Fc and HRP-conjugated goat anti-human IgG kappa light chain antibodies. FAM19A5 protein levels were measured using the LRRC4 family mimetic molecule-based sandwich ELISA assay described in Figure 1.

[0061] Figure 6 shows FAM19A5 protein levels in monkey blood at various time points after anti-FAM19A5 administration. Anti-FAM19A5 antibodies were administered at three different doses (3 mg / kg ("1"), 10 mg / kg ("2") or 30 mg / kg ("3")) and monkeys were administered a single antibody intravenously. FAM19A5 protein levels were measured using the LRRC4 family mimetic molecule-based sandwich ELISA assay described in Figure 1.

[0062] Figure 7 shows FAM19A5 protein levels in blood of traumatic brain injury (TBI)-induced mice treated with anti-FAM19A5 antibody. Mice were administered anti-FAM19A5 ("2") antibody twice (see arrows) (30 mg / kg per dose). Untreated TBI-induced mice and TBI-induced mice administered human IgG antibody ("1") served as control groups. FAM19A5 protein levels were measured using the LRRC4 family mimetic molecule-based sandwich ELISA assay described in Figure 1. Data are shown as mean ± SEM.

[0063] [Mode for carrying out the invention] The present invention provides mimetic molecules (e.g., polypeptides and small molecules) that can inhibit, reduce and / or dissociate the binding between FAM19A5 protein and LRRC4 protein family members (i.e., LRRC4 family mimetic molecules). As described herein, the LRRC4 family mimetic molecules of the present invention are similar to the LRRC4 protein family members and therefore can compete for binding to FAM19A5 protein. In some aspects, the LRRC4 family mimetic molecules described herein exhibit one or more properties (e.g., increased binding affinity and / or stability) and can compete with naturally occurring LRRC4 protein family members for binding to FAM19A5 protein. Thus, the present application discloses for the first time that such LRRC4 family mimetic molecules can be used to confirm the expression level of FAM19A5 protein in a subject. Compared to other approaches available in the art, the LRRC4 family mimetic molecules of the invention can be used to more accurately ascertain FAM19A5 protein levels (e.g., in biological fluids such as plasma, serum, or cerebrospinal fluid). Additional aspects of the invention are provided throughout this application.

[0064] To facilitate understanding of the subject matter disclosed herein, a number of terms and phrases are defined. Additional definitions are provided throughout the detailed description.

[0065] I. Definition Throughout this specification, the term "a" or "an" entity means one or more of that entity; for example, "a molecule" is understood to refer to one or more molecules. Thus, the terms "a," "one or more," and "at least one" may be used interchangeably herein.

[0066] Also, "and / or" as used herein should be considered as a specific disclosure of each of the two specified features or components without the other. Thus, the term "and / or" as used herein in phrases such as "A and / or B" is intended to include "A and B," "A or B," "A alone" and "B alone." Similarly, the term "and / or" as used in phrases such as "A, B and / or C" is intended to include each of the following aspects: A, B and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A alone; B alone; and C alone.

[0067] Where the term "comprising" is used herein, it is understood that similar aspects described with the terms "consisting of" and / or "essentially consisting of" are also provided.

[0068] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by a person of ordinary skill in the art related to the present invention. For example, references (The Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; Cell and Molecular Biology, 3rd ed., 1999, Academic Press; and the Oxford Dictionary Of Biochemistry And Molecular Biology, Revised, 2000, Oxford University Press) provide those skilled in the art with the common dictionary meaning of many terms used herein.

[0069] Units, prefixes, and symbols are expressed in SI (System International de Unites) accepted format. Numeric ranges are inclusive of the numbers defining the range. Unless otherwise indicated, amino acid sequences are written from left to right in the amino to carboxy orientation. The headings provided herein are not intended to limit the various aspects of the invention, which may have been had by reference to the specification as a whole. Thus, the terms defined below are more fully defined by reference to the specification as a whole.

[0070] The term "about" is used herein to mean approximately, in the region, roughly, etc. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the stated numerical values. In general, the term "about" may modify a numerical value above or below (higher or lower) the stated value by, for example, a variance of 10%.

[0071] As used herein, the term "alkenyl" refers to a group containing hydrogen and carbon and containing at least one carbon-carbon double bond.

[0072] The term "alkoxy," as used herein, refers to an alkyl group attached to the parent molecular moiety through an oxygen atom.

[0073] As used herein, the term "alkyl" refers to a group that contains hydrogen and carbon and no double bonds.

[0074] The term "alkynyl" as used herein refers to a group containing hydrogen and carbon and containing at least one carbon-carbon triple bond.

[0075] The term "amino" as used herein means --NH.sub.2.

[0076] The term "bicycloalkenyl" as used herein means a fused, spirocyclic or bridged bicyclic hydrocarbon ring system containing at least one double bond.

[0077] The term "bicycloalkyl" as used herein means a fused, spirocyclic or bridged bicyclylcycloalkyl ring.

[0078] The term "cycloalkenyl" as used herein means an unsaturated non-aromatic monocyclic hydrocarbon ring system having 0 heteroatoms. Representative examples of cycloalkenyl groups include, but are not limited to, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl.

[0079] The term "cycloalkyl" as used herein means a saturated monocyclic hydrocarbon ring system having 0 heteroatoms. Representative examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0080] The term "formyl" as used herein means --CHO.

[0081] The terms "halo" and "halogen" as used herein mean Cl, Br, I, or F.

[0082] The term "haloalkoxy," as used herein, refers to a haloalkyl group attached to the parent molecular moiety through an oxygen atom.

[0083] The term "haloalkyl," as used herein, refers to an alkyl group substituted with one, two, three, or four halogen atoms.

[0084] The term "heteroaryl" as used herein means an aromatic ring containing 1, 2, or 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. Representative examples of heteroaryl groups include, but are not limited to, furyl, imidazolyl, pyrazolyl, pyridinyl, pyrrolyl, thiazolyl, and thienyl.

[0085] The term "heterobicycloalkyl" as used herein means a non-aromatic bicyclic ring system containing 1, 2, 3, or 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and optionally containing one or more double bonds. The heterobicycloalkyl groups of the present invention can be attached to the parent molecular moiety through any carbon or nitrogen atom in the group.

[0086] The term "heterocycloalkyl" as used herein refers to a non-aromatic ring containing 1, 2, 3, or 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and optionally containing one or more double bonds. The heterocycloalkyl groups of the present invention can be attached to the parent molecular moiety through any carbon or nitrogen atom in the group. Representative examples of heterocycloalkyl groups include, but are not limited to, morpholinyl, piperazinyl, piperidinyl, tetrahydrofuranyl, tetrahydropyranyl, and thiomorpholinyl.

[0087] The term "leucine-rich repeat-containing 4 protein family" or "LRRC4 protein family" (including derivatives thereof) refers to a family of proteins described as key synaptic organizers and playing a role in various stages of neural circuit formation, including neuronal migration, neurite outgrowth, formation of synaptic contacts, and functional assembly (see, e.g., Woo et al., Mol Cell Neurosci 42(1):1-10 (September 2009)). The LRRC4 protein family (referred to herein as "LRRC4 protein family members" or "LRRC4 protein family members" (or derivatives thereof)) includes three members: (1) LRRC4, (2) LRRC4B, and (3) LRRC4C. LRRC4 protein family members generally contain nine leucine-rich repeat (LRR) domains flanking the LRR N- and C-terminus (see FIG. 3A). Such LRR domains are known to interact with the fibronectin type III domain of presynaptic receptor protein tyrosine phosphatase (RPTP) proteins (see, for example, Won et al., Mol Cells 41(7):622-630 (Jul. 2018)). The LRR domain is followed by an immunoglobulin-like C2 type (IG) and a threonine (Thr)-rich domain, which together form the extracellular portion of the LRRC4 protein family members. Unlike other members, the LRRC4B protein has an extra glycine (Gly)-rich domain between the IG and Thr-rich domains. In addition to the extracellular portion, the LRRC4 protein family members further contain a transmembrane (TM) domain and a postsynaptic density-binding (PB) domain at the C-terminus of the protein.

[0088] In humans, the gene encoding the LRRC4 protein is located on chromosome 7 (nucleotides 128,027,071-128,032,107 of GenBank Accession Number NC_000007.14; minus strand orientation). Synonyms for the LRRC4 protein are known, non-limiting examples include: "Nasopharyngeal Carcinoma-Associated Gene 14 Protein," "Brain Tumor-Associated Protein BAG," "Netrin-G2 Ligand," "NAG14," "NGL-2," and "BAG." The amino acid sequence of the LRRC4 protein is 653 amino acids in length and is provided in Table 1 (below). The full length ectodomain of the LRRC4 protein corresponds to amino acid residues 39-527 of SEQ ID NO:1 (i.e., SEQ ID NO:4). Unless otherwise indicated, the term "LRRC4 protein" (including its synonyms) includes any variant or isoform of the LRRC4 protein that is naturally expressed by a cell.

[0089] [Table 1]

[0090] In humans, the gene encoding the LRRC4B protein is located on chromosome 19 (nucleotides 50,516,892-50,568,435 of GenBank Accession Number NC_000019.10; minus strand orientation). Synonyms for the LRRC4B protein are known, and non-limiting examples include: "Netrin-G3 ligand," "LRIG4," "NGL-3," "HSM," and "DKFZp761A179." The amino acid sequence for the LRRC4B protein is 713 amino acids long and is provided in Table 2 (below). The full-length ectodomain of the LRRC4B protein corresponds to amino acid residues 36-576 of SEQ ID NO:2 (i.e., SEQ ID NO:5). Unless otherwise specified, the term "LRRC4B protein (including synonyms thereof)" includes any variant or isoform of the LRRC4B protein that is naturally expressed by a cell.

[0091] [Table 2]

[0092] In humans, the gene encoding the LRRC4C protein is located on chromosome 11 (nucleotides 40,107,066-41,460,419 of GenBank Accession Number NC_000011.10; minus strand orientation). Synonyms of the LRRC4C protein are known, and non-limiting examples include "NGL-1", "Netrin-G1 ligand" and "KIAA1580". The amino acid sequence for the LRRC4C protein is 640 amino acids in length and is provided in Table 3 (below). The full length ectodomain of the LRRC4C protein corresponds to amino acids 45-527 of SEQ ID NO:3 (i.e., SEQ ID NO:6). Unless otherwise indicated, the term "LRRC4C protein (including synonyms thereof)" includes any variant or isoform of the LRRC4C protein that is naturally expressed by a cell.

[0093] [Table 3]

[0094] As used herein, the term "FAM19A5 binding domain" refers to a segment / fragment of a LRRC4 protein family member capable of binding to a FAM19A5 protein.

[0095] The term "family with sequence similarity 19, member A5" or "FAM19A5" refers to a protein that belongs to the TAFA family of five highly homologous proteins (also known as the FAM19 family) and is expressed primarily in the brain and spinal cord. FAM19A5 is also known as "TAFA5" or "chemokine-like protein TAFA-5."

[0096] In humans, the gene encoding FAM19A5 is located on chromosome 22. There are multiple human FAM19A5 (UniProt:Q7Z5A7) isoforms that are said to be produced by alternative splicing. Isoform 1 (UniProt:Q7Z5A7-1) consists of 132 amino acids, isoform 2 (UniProt:Q7Z5A7-2) consists of 125 amino acids, and isoform 3 (UniProt:Q7Z5A7-3) consists of 53 amino acids. Human FAM19A5 protein is said to exist in membrane-bound and water-soluble (secreted) forms. Isoform 1 is considered a membrane protein with one transmembrane domain. Isoform 2, reported in the literature (Tang TY et al., Genomics 83(4):727-34(2004)) as a secreted protein (water-soluble), contains a signal peptide at amino acid positions 1-25. Isoform 1 is considered a membrane protein and is predicted based on EST data. Table 4 (below) provides the amino acid sequences of the three known human FAM19A5 isoforms. Unless otherwise specified, "FAM19A5" includes any variant or isoform of the FAM19A5 protein that is naturally expressed by a cell. Thus, in some aspects, a polypeptide described herein (e.g., comprising a FAM19A5 binding domain of an LRRC4 protein family member) can inhibit binding of FAM19A5 isoform 1, isoform 2, and / or isoform 3 to an LRRC4 protein family member.

[0097] [Table 4]

[0098] The term "endogenous" as used to describe a member of the LRRC4 protein family refers to an LRRC4 family protein that is naturally present in a subject. As described herein, the mimetic molecules of the present invention differ (structurally and / or functionally) from an endogenous LRRC4 protein family member.

[0099] "Binding affinity" generally refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an LRRC4 mimetic molecule) and its binding partner (e.g., a FAM19A5 protein). Unless otherwise specified, "binding affinity" as used herein refers to the intrinsic binding affinity reflecting a 1:1 interaction between members of a binding pair. The affinity of a molecule X (e.g., a mimetic molecule described herein that contains a FAM19A5 binding domain of an LRRC4 protein family member) for its partner Y (e.g., FAM19A5) is generally measured by the dissociation constant (K D Affinity can be expressed as the equilibrium dissociation constant (K D ) and the equilibrium association constant (K A ), which may be measured and / or expressed in a number of ways known in the art, including but not limited to. D is k off / k on and expressed as molar concentration (M), whereas K A is k on / k off It is calculated as the quotient of k on means, for example, the binding rate constant of an antibody to an antigen, and k off means, for example, dissociation of an antibody against an antigen. on and k off may be measured by techniques known to those of skill in the art such as immunoassays (eg, enzyme-linked immunosorbent assay (ELISA)), BIACORE, or kinetic exclusion assay (KinExA).

[0100] As used herein, the terms "specifically bind," "specifically recognize," "specific binding," "selective binding," and "selectively bind" are similar terms and refer to the binding of a molecule (e.g., an LRRC4 family mimetic molecule) to an antigen (e.g., a FAM19A5 protein), as would be understood by one of skill in the art. For example, a molecule that specifically binds to an antigen is generally capable of binding to other peptides or polypeptides with a lower affinity, as measured, for example, by immunoassays, BIACORE, KinExA3000 instruments (Sapidyne Instruments, Boise, ID), or other known assays. In some aspects, a molecule that specifically binds to an antigen exhibits a K A At least about 2 log, at least about 2.5 log, at least about 3 log, at least about 4 log or greater than A The antigen binds to the antigen having the formula:

[0101] The term "antigen" as used herein means any natural or synthetic immunogenic substance such as a protein, peptide or hapten. As is evident from the present invention, the antigen may be the FAM19A5 protein or a fragment thereof.

[0102] A molecule that "competes with other proteins for binding to a target" (e.g., an "LRRC4 family mimetic molecule") refers to a molecule that inhibits (partially or completely) the binding of other proteins (e.g., naturally occurring LRRC4 protein family members) to a target. Whether two compounds compete with each other for binding to a target, i.e., whether and to what extent a LRRC4 family mimetic molecule described herein inhibits the binding of a naturally occurring member of the LRRC4 protein family to FAM19A5 protein, may be determined using known competition experiments. In some aspects, the LRRC4 mimetic molecule described herein competes to inhibit the binding of a naturally occurring member of the LRRC4 protein family to FAM19A5 protein by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 100%. Competitive assays may be performed as described herein or, for example, in the literature (Ed Harlow and David Lane, Cold Spring Harb Protoc; 2006; doi:10.1101 / pdb.prot4277 or Chapter 11 of "Using Antibodies" by Ed Harlow and David Lane, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA).

[0103] Other competitive binding assays that can be used with the present invention include solid-phase direct or indirect radioimmunoassays (RIA), solid-phase direct or indirect enzyme immunoassays (EIA), sandwich competition assays (see Stahli et al., Methods in Enzymology 9:242 (1983)); solid-phase direct biotin-avidin EIA (see Kirkland et al., J. Immunol. 137:3614 (1986)); solid-phase direct label assays, solid-phase direct label sandwich assays (see Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Press (1988)); solid-phase direct label RIA using 1-125 labels (see Morel et al., Mol. Immunol. 25(1):7 (1988)); solid-phase direct biotin-avidin EIA (see Cheung et al., J. Immunol. 25(1):7 (1988)); al., Virology 176:546 (1990)); and directly labeled RIA. (see Moldenhauer et al., Scand. J. Immunol. 32:77 (1990)).

[0104] As used herein, the term "naturally occurring" or "naturally occurring" refers to the fact that an object (e.g., a protein) can be found in nature. For example, a polypeptide or polynucleotide sequence that is present in an organism (including viruses) that may be isolated from a natural source and has not been intentionally modified by man in a laboratory is naturally occurring. As further described elsewhere herein, the LRRC4 family mimetic molecules useful in the present invention are not naturally occurring.

[0105] A "mimetic molecule" refers to a molecule that is similar in structure and / or function to another molecule (a "reference molecule"). For example, in some aspects, a mimetic molecule can share a partial structure or sequence with a reference molecule, and such a mimetic molecule can exhibit one or more characteristics of the reference molecule. However, as will be apparent from the present invention, structural or sequence similarity is not always required. In some aspects, a mimetic molecule can differ in structure but perform similarly to the reference molecule. As described herein, in some aspects, a mimetic molecule includes a small molecule. In some aspects, a mimetic molecule includes a peptide. In some aspects, a mimetic molecule is not an antibody or an antigen-binding portion thereof.

[0106] A "polypeptide" refers to a chain containing at least two contiguously linked amino acid residues, with no upper limit to the length of the chain. One or more amino acid residues of a protein may contain modifications such as, but not limited to, glycosylation, phosphorylation, or disulfide bond formation. A "protein" may include one or more polypeptides.

[0107] The term "nucleic acid" or "nucleic acid molecule" as used herein is intended to include DNA molecules and RNA molecules. A nucleic acid molecule may be single-stranded or double-stranded and may be cDNA.

[0108] The term "vector" as used herein is intended to mean a nucleic acid molecule capable of transporting additional nucleic acids to which it has been linked. One type of vector is a "plasmid," which refers to a circular double stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, into which additional DNA segments can be ligated into the viral genome. Certain vectors (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors) are capable of autonomous replication in a host cell into which they are introduced. Other vectors (e.g., non-episomal mammalian vectors) may be integrated into the genome of the host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Certain vectors are also capable of directing the expression of genes to which they are operably linked. Such vectors are referred to herein as "recombinant expression vectors (or simply "expression vectors")." In general, expression vectors useful in recombinant DNA techniques are often in the form of plasmids. As used herein, "plasmid" and "vector" may be used interchangeably as the plasmid is the most commonly used form of vector. However, other forms of expression vectors, such as viral (eg, replication defective retroviruses, adenoviruses and adeno-associated viruses) vectors, which serve equivalent functions are also included.

[0109] The term "recombinant host cell" (or simply "host cell"), as used herein, is intended to refer to a cell that contains a nucleic acid that is not naturally present in the cell, and may be a cell into which a recombinant expression vector has been introduced. It should be understood that such terms are intended to refer to not only the particular subject cell, but also the progeny of such a cell. Because certain modifications may occur in successive generations due to mutation or environmental influences, such progeny may not actually be identical to the parent cell, but are still included within the scope of the term "host cell" as used herein.

[0110] As used herein, the term "administering" refers to the physical introduction of an agent (e.g., an LRRC4 mimetic molecule) or a composition containing an agent to a subject using any of a variety of methods and delivery systems known to those of skill in the art. Non-limiting examples of administration routes available in the prior art include intravenous, intraperitoneal, intramuscular, subcutaneous, spinal or other parenteral administration routes, such as injection or infusion. The term "parenteral administration," as used herein, generally refers to modes of administration other than enteral and topical administration by injection, and includes, but is not limited to, intravenous, intraperitoneal, intramuscular, intraarterial, intracheal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, as well as in vivo electroporation. Alternatively, the molecules described herein (e.g., the LRRC4 family mimetic molecules described herein) may be administered via a parenteral route, such as a topical, epidermal or mucosal route of administration, e.g., intranasally, orally, vaginally, rectally, sublingually or topically. Administration may also be, for example, once, multiple times and / or over one or more extended periods of time.

[0111] As used herein, the term "subject" includes any human or non-human animal. The term "non-human animal" includes all vertebrates, e.g., mammals and non-mammals such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc.

[0112] As used herein, the term "neuron" includes electrically excitable cells that process and transmit information by electrical and chemical signals. Neurons are the primary components of the brain and spinal cord of the CNS and the ganglia of the peripheral nervous system (PNS) and can be connected to each other to form a neuronal network. A typical neuron is composed of a cell body (soma), dendrites, and an axon. The soma of a neuron contains the nucleus. The dendrites of a neuron are cell extensions with numerous branches from which most input to the neuron originates. Axons are finer cable-like processes that extend from the cell body and transmit neural signals away from the cell body and transmit certain types of information back to the cell body.

[0113] As used herein, the term "therapeutically effective amount" refers to an amount of a substance (e.g., an LRRC4 mimetic molecule described herein) that is effective, alone or in combination with other therapeutic agents, to "treat" a disease or disorder, which reduces or alleviates the risk, latency, likelihood or occurrence of a disease or disorder (e.g., a neurological disease described herein). A "therapeutically effective amount" includes an amount of a substance or therapeutic agent that provides some improvement or benefit to a subject having or at risk of having a disease or disorder (e.g., a neurological disease described herein). Thus, a "therapeutically effective amount" is an amount that reduces or provides partial relief, alleviation of the risk, latency, likelihood or occurrence of a disease, and / or reduces at least one indicator and / or provides a reduction in at least one clinical symptom of the disease or disorder.

[0114] As used herein, the term "diagnosis" (or its derivatives) refers to a method available for identifying or predicting whether a patient suffers from a given disease or condition and identifying subjects suitable for treatment. One of skill in the art can diagnose based on one or more diagnostic markers (e.g., FAM19A5), the presence, absence, amount, or change in amount of said diagnostic markers indicating the presence, severity, or absence of a pathology. In some aspects, an increase in FAM19A5 protein expression (e.g., in a biological sample of a subject) is evidence of a disease or pathology. Non-limiting examples of such diseases or disorders are provided herein. The term "diagnosis" does not imply the ability to determine with 100% accuracy the presence or absence of a particular disease or disorder, nor does it imply that a given course or outcome is more likely to occur than not. Instead, one of skill in the art will understand that the term "diagnosis" refers to an increased likelihood that a particular disease or disorder exists in a subject.

[0115] II. Diagnostic Methods The present invention provides a method for determining the expression level of FAM19A5 in a subject in need thereof using an LRRC4 family mimetic molecule. As identified herein, in some aspects, the method for determining the expression level of FAM19A5 protein comprises: (i) contacting an LRRC4 family mimetic molecule (e.g., as described herein) with a biological sample obtained from the subject, and (ii) measuring the expression level of FAM19A5 protein in the biological sample. In some aspects, the method for determining the expression level of FAM19A5 protein comprises measuring the expression level of FAM19A5 protein in a biological sample obtained from the subject, the biological sample being contacted with an LRRC4 family mimetic molecule.

[0116] As is evident from the present invention, contacting the biological sample with the LRRC4 family mimetic molecule causes the LRRC4 family mimetic molecule to specifically bind to FAM19A5 protein present in the biological sample. As described herein, in some aspects, the LRRC4 family mimetic molecule that binds to FAM19A5 protein with greater binding affinity is compared to other binding agents available in the art (e.g., natural or synthetic). Thus, in some aspects, when the biological sample contains other FAM19A5 binding agents (e.g., naturally occurring LRRC4 protein family members), the LRRC4 mimetic molecule described herein can compete with other FAM19A5 binding agents for binding to FAM19A5 protein. In some aspects, upon contact with a biological sample containing FAM19A5 protein, the LRRC4 family mimetic molecules described herein bind to at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% of the FAM19A5 protein present in the biological sample. In some aspects, the LRRC4 family mimetic molecules of the present invention bind to all of the FAM19A5 protein present in the biological sample.

[0117] Without being bound to any one theory, in some aspects, the increased binding affinity of the LRRC4 family mimetic molecule to FAM19A5 protein provides greater accuracy when determining FAM19A5 protein levels in a biological sample of a subject. For example, many detection assays use antibodies or Fc fragments to determine and measure the level of a protein of interest (i.e., target protein) in a sample (FIGS. 1A and 1B). However, such assays often have high background due to, for example, Fc-mediated non-specific binding and / or competition in binding to the target protein with antibodies naturally present in the sample. As confirmed herein, the LRRC4 family mimetic molecule of the present invention exhibits certain properties (e.g., lacking an Fc fragment and binding to FAM19A5 protein with high affinity) and when used to determine FAM19A5 protein levels in a sample (FIG. 1C), the background is reduced and FAM19A5 protein levels can be measured more accurately. As used herein, the term "accuracy" (and its derivatives) refers to the ability to closely measure (e.g., within ±10%) the actual amount of a substance (e.g., FAM19A5 protein) present in a given sample, which may be the result of, for example, reduced non-specific (i.e., background) signal.

[0118] Thus, in some aspects, the accuracy of measuring FAM19A5 protein expression levels in a biological sample using a method described herein (e.g., comprising contacting the biological sample with an LRRC4 family mimetic molecule) is increased compared to a control method (e.g., a corresponding method that does not include contacting the biological sample with an LRRC4 family mimetic molecule described herein, e.g., a method that comprises contacting the biological sample with an anti-FAM19A5 antibody). In some aspects, the accuracy is increased by at least about 1-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, or at least about 50-fold compared to the control method.

[0119] After contacting the biological sample with a LRRC4 family mimetic molecule as described herein, the method includes measuring the expression level of FAM19A5 protein in the biological sample. In some aspects, the measuring step includes quantifying the amount of LRRC4 family mimetic molecule bound to FAM19A5 protein (i.e., "LRRC4 family mimetic molecule-FAM19A5 complex"). Any suitable method / assay in the art may be used to quantify the amount of LRRC4 family mimetic molecule-FAM19A5 complex in the sample. Non-limiting examples include ELISA, immunohistochemistry, Western blot, radioimmunoassay, radical-immunodiffusion, immunoprecipitation, Ouchterlony immunodiffusion, rocket immunoelectrophoresis, tissue immunostaining, complement fixation, FACS, protein chips, Meso Scale Discovery (MSD) ELISA-based assay, SIMOA technology, or combinations thereof. In some aspects, the amount of LRRC4 family mimetic-FAM19A5 complex is quantified using Mesoscale Discovery (MSD) ELISA-based analysis. In some aspects, the amount of LRRC4 family mimetic-FAM19A5 complex is quantified using SIMOA technology. In some aspects, the amount of LRRC4 family mimetic-FAM19A5 complex is quantified using ELISA analysis. In some aspects, the ELISA analysis comprises a sandwich ELISA analysis as depicted in FIG. 1C.

[0120] As will be apparent from the present invention, the above-described methods can be used for a wide range of diagnostic and / or therapeutic purposes. For example, the applicant has determined that certain diseases or disorders may be associated with abnormal (e.g., increased) levels of FAM19A5 expression and / or activity (see, e.g., US11,155,613; US2020 / 0157202; US2020 / 0300870; US2021 / 0070849; WO2020 / 079595; US2020 / 0223914; US2020 / 0299373; and WO2020 / 136603, each of which is incorporated herein by reference in its entirety). Thus, in some aspects, the invention provides methods of diagnosing a disease or disorder in a subject in need thereof, comprising: (i) contacting a biological sample obtained from the subject with an LRRC4 family mimetic molecule (e.g., as described herein); and (ii) measuring the expression level of FAM19A5 protein in the described biological sample. In some aspects, the method of diagnosing a disease or disorder comprises measuring the expression level of FAM19A5 protein in the biological sample obtained from the subject after contacting the biological sample with the LRRC4 family mimetic molecule.

[0121] In some aspects, an increased expression level of FAM19A5 protein compared to a control group indicates that the subject suffers from or is at risk of developing a disease or disorder (e.g., associated with increased FAM19A5 expression and / or activity). In some aspects, the control group comprises a corresponding expression level in a biological sample from a subject prior to suffering from or being at risk of developing the disease or disorder. In some aspects, the control group comprises an expression level of FAM19A5 protein in a biological sample obtained from a corresponding subject suffering from or not at risk of developing the disease or disorder.

[0122] In some aspects, the expression level of FAM19A5 protein in a subject's biological sample is increased by at least about 1-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold compared to a control group.

[0123] As described herein, the methods (e.g., for measuring expression levels of FAM19A5 protein and / or diagnosing a disease or disorder) include contacting a biological sample obtained from a subject with a LRRC4 family mimetic molecule. As used herein, the term "biological sample" refers to any sample containing a substance that may be derived from a subject (e.g., a human). Non-limiting examples of biological samples useful in the present invention include: blood, cerebral spinal fluid (CSF), serum, plasma, tissue, cell culture medium, saliva, urine, or combinations thereof. In some aspects, the biological sample comprises blood. In some aspects, the biological sample comprises CSF. In some aspects, the biological sample comprises serum. In some aspects, the biological sample comprises plasma. In some aspects, the biological sample comprises cell culture medium. In some aspects, the biological sample comprises both blood and CSF. In some aspects, the biological sample comprises any combination of blood, CSF, serum, plasma, and culture medium. In some aspects, the expression level of FAM19A5 protein determined from the biological sample correlates to the total expression level of FAM19A5 protein in the subject.

[0124] As further described in the present invention, the applicant has previously determined that administration of a FAM19A5 antagonist (e.g., an anti-FAM19A5 antibody) can be useful in treating diseases or disorders associated with increased expression levels of FAM19A5 protein.

[0125] Thus, in some aspects, the present invention relates to a method for identifying a subject suitable for treatment with a FAM19A5 antagonist, the method comprising: (i) contacting a biological sample obtained from the subject with a LRRC4 family mimetic molecule; and (ii) measuring the expression level of FAM19A5 protein in the biological sample. As described herein, in some aspects, the measuring step comprises quantifying the amount of LRRC4 family mimetic molecule bound to FAM19A5 protein. Any suitable method / analysis (e.g., as described herein) may be used to quantify the amount of LRRC4 family mimetic-FAM19A5 complex present in the sample. In some aspects, the method for identifying a subject suitable for treatment with a FAM19A5 antagonist comprises measuring the expression level of FAM19A5 protein in a biological sample obtained from the subject after contacting the biological sample with a LRRC4 family mimetic molecule.

[0126] In some aspects, if the expression level of FAM19A5 protein in the biological sample is increased compared to a control group, the subject is suitable for treatment with a FAM19A5 antagonist. In some aspects, the control group comprises a corresponding expression level in a biological sample from a subject prior to suffering from or being at risk for developing the disease or disorder. In some aspects, the control group comprises an expression level of FAM19A5 protein in a biological sample obtained from a corresponding subject not suffering from or at risk for developing the disease or disorder.

[0127] In some aspects, the expression level of FAM19A5 protein in a biological sample of a subject suitable for treatment with a FAM19A5 antagonist is increased by at least about 1-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold compared to a control group.

[0128] III. Treatment method In some aspects, the invention provides methods of treating a disease or disorder in a subject. For example, in some aspects, the subject has been identified or diagnosed as suffering from or at risk of developing a disease or disorder using a method described herein (e.g., contacting a biological sample from the subject with a LRRC4 family mimetic molecule and measuring the expression level of FAM19A5 protein in the biological sample). In some aspects, the subject has been identified as suitable for treatment, e.g., using a method described herein.

[0129] Thus, in some aspects, the invention provides a method of treating a disease or disorder associated with an increased expression level of FAM19A5 protein in a subject in need thereof, the method comprising measuring the expression level of FAM19A5 protein in a biological sample obtained from the subject by contacting the biological sample with a LRRC4 family mimetic molecule (e.g., as described herein), the subject having been administered a therapeutic agent for the disease or disorder prior to the measuring step. In some aspects, the measuring step comprises quantifying the amount of LRRC4 family mimetic molecule bound to the FAM19A5 protein. As further described in the invention, any suitable method / assay known in the art and / or described herein may be used to quantify the amount of LRRC4 family mimetic-FAM19A5 complex in the sample.

[0130] In some aspects, the expression level of FAM19A5 protein in the subject's biological sample is increased compared to a control group. In some aspects, the control group includes a corresponding expression level in a subject's biological sample before suffering from or at risk of developing the disease or disorder. In some aspects, the control group includes a corresponding expression level of FAM19A5 protein in a biological sample obtained from a corresponding subject suffering from or not at risk of developing the disease or disorder. In some aspects, the expression level of FAM19A5 protein in the subject's biological sample is increased by at least about 1-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold compared to the control group.

[0131] In some aspects, if the expression level of FAM19A5 protein is increased compared to a control group, the method of treating a disease or disorder provided herein further comprises administering to the subject an additional dose of a therapeutic agent. In some aspects, the additional dose is the same as the amount previously administered to the subject. In some aspects, the additional dose is greater than the amount previously administered to the subject. In some aspects, the additional dose is less than the amount previously administered to the subject. In some aspects, the additional dose is administered at the same dosing interval as previously administered to the subject. In some aspects, the additional dose is administered to the subject at a different dosing interval (e.g., longer or shorter) from one another. In some aspects, the additional dose may be administered in combination with an additional therapeutic agent, non-limiting examples of which are provided elsewhere herein.

[0132] In some aspects, when the expression level of the FAM19A5 protein increases compared to a control group, the method includes discontinuing treatment. When treatment is discontinued, in some aspects, the method includes administering another therapeutic agent (e.g., another FAM19A5 antagonist) to the subject.

[0133] In some aspects, the level of FAM19A5 protein is the same as compared to a control group (e.g., the expression level of FAM19A5 protein in a biological sample obtained from a corresponding subject not suffering from or at risk of developing a disease or disorder). In some aspects, the expression level of FAM19A5 protein is the same as that of a control group, and the expression level of FAM19A5 protein is at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% of the corresponding expression in the control group. Thus, when comparing the amount of FAM19A5 protein in a sample to that of a control group, the term "same" includes both an amount that matches the amount present in the control group (i.e., 100% match) and is nearly as close (e.g., within about 20%).

[0134] In some aspects, when the expression level of FAM19A5 protein is similar compared to a control group, the methods of treating a disease or disorder provided herein further comprise administering to the subject an additional dose of a therapeutic agent. In some aspects, the additional dose is the same as the dose previously administered to the subject. In some aspects, the additional dose is greater than the dose previously administered to the subject. In some aspects, the additional dose is less than the dose previously administered to the subject. In some aspects, the additional dose is administered to the subject at the same dosing interval as previously administered to the subject. In some aspects, the additional dose is administered to the subject at a different dosing interval (e.g., longer or shorter) from each other. In some aspects, the additional dose may be administered with an additional therapeutic agent (e.g., as described herein).

[0135] In some aspects, when the expression level of FAM19A5 protein is similar compared to the control group, the method includes discontinuing the treatment. When the treatment is discontinued, in some aspects, the method includes administering another therapeutic agent (e.g., another FAM19A5 antagonist) to the subject.

[0136] In some aspects, the expression level of FAM19A protein is decreased relative to a control group (e.g., the expression level of FAM19A5 protein in a biological sample obtained from a corresponding subject not suffering from or at risk of developing a disease or disorder). In some aspects, the expression level of FAM19A5 protein is decreased by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% in the subject's biological sample compared to the control group.

[0137] In some aspects, if the expression level of FAM19A5 protein decreases compared to the control group, the treatment is maintained (e.g., the subject continues the same treatment, e.g., at the same dose and / or dosing interval). In some aspects, if the expression level of FAM19A5 protein decreases, the treatment is altered. For example, in some aspects, the treatment is altered when the subject receives a lower dose of the therapeutic agent. In some aspects, the treatment is altered when the subject receives the same treatment (e.g., at the same dose), but with a longer dosing interval. In some aspects, the treatment is altered when the subject receives a lower dose of the therapeutic agent with a longer dosing interval.

[0138] As is apparent from at least the invention, in some aspects, the described subject matter may be useful for determining whether a subject will respond to a treatment for a disease or disorder described herein (e.g., associated with increased FAM19A5 protein expression). Thus, in some aspects, the invention relates to a method for identifying a subject that will respond to a treatment for a disease or disorder associated with increased expression levels of FAM19A5 protein, the method comprising measuring the expression level of FAM19A5 protein in a biological sample obtained from the subject by contacting the biological sample with a LRRC4 family mimetic molecule (e.g., as described herein), the subject having been administered a therapeutic agent prior to the measurement. In some aspects, the measuring step comprises quantifying the amount of LRRC4 family mimetic molecule bound to FAM19A5 protein. As described in the invention, any suitable method / assay known in the art and / or described herein may be used to quantify the amount of LRRC4 family mimetic molecule-FAM19A5 complex in the sample.

[0139] In some aspects, a subject has responded to treatment if the expression level of FAM19A5 protein is decreased compared to a control group (e.g., the expression level of FAM19A5 protein in a biological sample obtained from a corresponding subject not suffering from or at risk of developing a disease or disorder). In some aspects, the expression level of FAM19A5 protein is decreased by at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% compared to a control group. In some aspects, if a subject is determined to be responsive to treatment, treatment is maintained (e.g., at the same dose and / or dosing interval), modified (e.g., at a lower dose and / or at a longer dosing interval), or discontinued as described.

[0140] In some aspects, if the expression level of FAM19A5 protein is increased, the same or similar compared to a control group (e.g., the expression level of FAM19A5 protein in a biological sample obtained from a corresponding subject not suffering from or at risk of developing a disease or disorder), the subject is not responding to the treatment. In some aspects, if the subject is determined to be not responding to the treatment, the treatment is discontinued and another therapeutic agent is administered to the subject. In some aspects, if the subject is not responding to the treatment, the subject is treated with a higher dose of the therapeutic agent. In some aspects, if the subject is not responding to the treatment, the subject is treated with the same dose but with a shorter dosing interval. In some aspects, if the subject is not responding to the treatment, the subject is treated with a higher dose of the treatment and with a shorter dosing interval. In some aspects, if the subject is not responding to the treatment, the therapeutic agent may be administered to the subject along with an additional therapeutic agent (e.g., as described herein).

[0141] In some aspects, in any of the methods provided herein that include administering a therapeutic agent to a subject (e.g., for a disease or disorder associated with increased expression levels of FAM19A5 protein), the treatment includes a FAM19A5 antagonist. In some aspects, the treatment includes a LRRC4 family mimetic molecule. In some aspects, the treatment includes both a FAM19A5 antagonist and a LRRC4 family mimetic molecule.

[0142] In some aspects, FAM19A5 antagonists useful in the present invention include antisense oligonucleotides, siRNAs, shRNAs, miRNAs, dsRNAs, aptamers, PNAs, vectors containing the same, or any combination thereof. In some aspects, FAM19A5 antagonists include antibodies that specifically bind to FAM19A5 protein (herein referred to as "anti-FAM19A5 antibodies") or antigen-binding portions thereof, polynucleotides encoding anti-FAM19A5 antibodies, or vectors containing the polynucleotides. Non-limiting examples of such FAM19A5 antagonists are described, for example, in US2019 / 0300599; US2020 / 0299373; US2021 / 0054062; and WO2020 / 141452, the entire contents of which are incorporated herein by reference. Additional information regarding LRRC4 family mimetic molecules useful in the present invention is provided separately herein.

[0143] In some aspects, when administered to a subject, FAM19A5 antagonists useful in the present invention can inhibit, slow, reduce, reverse and / or prevent gliosis and its deleterious effects on the CNS of a subject. In some aspects, when administered to a subject, FAM19A5 antagonists can inhibit, slow, reduce, reverse and / or prevent excessive or abnormal proliferation of reactive astrocytes and its deleterious effects on the CNS of a subject. In some aspects, when administered to a subject, FAM19A5 antagonists can stimulate, promote, increase and / or activate neuronal growth in the subject. In some aspects, when administered to a subject, FAM19A5 antagonists can enhance and / or promote neuronal survival in the subject. In some aspects, when administered to a subject, FAM19A5 antagonists can enhance and / or promote axonal regrowth in the subject. In some aspects, when administered to a subject, a FAM19A5 antagonist can increase neurite outgrowth and / or synaptogenesis in neurons of the subject.

[0144] In some aspects, when administered to a subject, the LRRC4 family mimetic molecules described herein can inhibit, reduce and / or dissociate the interaction between the FAM19A5 protein and a member of the LRRC4 protein family. In some aspects, after administration, complex formation between the FAM19A5 protein and a member of the LRRC4 family mimetic molecule is reduced by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% compared to control formation (e.g., a corresponding value in a pre-administration subject or a corresponding value in a non-administered subject).

[0145] As described in the present invention, the binding of FAM19A5 protein to LRRC4 protein family members can suppress the activity of LRRC4 protein family members. For example, in some aspects, the formation of FAM19A5-LRRC4 family protein complexes can induce impaired neural circuit formation, resulting in an imbalance in the dynamic gain and loss of synapses that is essential for the healthy function of central and peripheral nervous systems.

[0146] Thus, in some aspects, a decrease in the formation of a complex between the FAM19A5 protein and a LRRC4 protein family member can increase the activity of the LRRC4 protein family member. In some aspects, after administration, the activity of the LRRC4 protein family member is increased by at least about 0.5-fold, at least about 1-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold compared to a control group (e.g., a corresponding value in a pre-administration subject or a corresponding value in a non-administered subject). Non-limiting examples of such activities can include neurite outgrowth, neuronal migration, and the formation and functional assembly of synaptic contacts.

[0147] Without being bound to any one theory, in some aspects, the therapeutic efficacy of one or more of the above-mentioned FAM19A5 antagonists and / or LRRC4 family mimetic molecules may be useful for treating or reducing one or more symptoms of a disease or disorder described herein (e.g., associated with increased FAM19A5 expression), including, but not limited to, amyotrophic lateral sclerosis (ALS), Alzheimer's disease, glaucoma, diabetic retinopathy, neuropathic pain, spinal cord injury, traumatic brain injury, stroke, Parkinson's disease, or a combination thereof.

[0148] In some aspects, the present invention provides a method of treating ALS in a subject in need thereof, comprising administering to the subject a therapeutic agent for ALS and measuring the expression level of FAM19A5 protein by contacting a biological sample from the subject with an LRRC4 family mimetic molecule described herein. In some aspects, the therapeutic agent for ALS comprises a FAM19A5 antagonist, an LRRC4 family mimetic molecule, or both. In some aspects, ALS that may be treated by the present invention includes sporadic ALS, familial ALS, or both. As used herein, the term "sporadic" ALS refers to ALS where the occurrence of ALS is not associated with any family dynamics. Approximately 90% or more of ALS diagnoses are associated with sporadic ALS. As used herein, the term "familial" ALS refers to ALS occurring more than once in a family, implying a genetic component to the disease. In some aspects, ALS that may be treated by the present invention includes primary lateral sclerosis (PLS). PLS can affect the upper motor neurons of the arms and legs. However, lower motor neuron signs develop within 4 years of onset in more than 75% of people with overt PLS, so PLS cannot be clearly diagnosed until that time. PLS has a better prognosis than classical ALS because it progresses more slowly, causes less functional loss, does not affect breathing ability, and does not cause severe weight loss. In some aspects, ALS includes progressive muscular atrophy (PMA). PMA can affect the lower motor neurons of the arms and legs. Although PMA is associated with a longer survival on average compared to classical ALS, it still progresses to other spinal cord regions over time, eventually leading to respiratory failure and death. Upper motor neuron signs can develop later in the PMA process, in which case the diagnosis may be changed to classical ALS.

[0149] In some aspects, the present invention provides a method of treating Alzheimer's disease in a subject in need thereof, the method comprising administering to the subject a therapeutic agent for Alzheimer's disease and measuring the expression level of FAM19A5 protein by contacting a biological sample from the subject with an LRRC4 family mimetic molecule as described herein. In some aspects, the therapeutic agent for Alzheimer's disease comprises a FAM19A5 antagonist, an LRRC4 family mimetic molecule, or both. Without being bound to any one theory, in some aspects, the method of treating Alzheimer's disease comprises reducing amyloid beta (Aβ) plaque burden in a subject (suffering from Alzheimer's disease). As used herein, "amyloid beta plaque" refers to all forms of abnormal deposition of amyloid beta, including large aggregates and small associations of several amyloid beta peptides, and may include any variant of amyloid beta peptide. Amyloid beta (Aβ) plaques are known to cause neuronal changes such as synaptic organization, synaptic morphology, synaptic density, synaptic conductivity loss, dendritic diameter changes, dendritic length changes, spine density changes, spine area changes, spine length changes, or spine head diameter changes. In some aspects, an increase in Aβ plaque load can cause synaptic loss in neurons.

[0150] In some aspects, provided herein is a method of treating glaucoma in a subject in need thereof, comprising administering to the subject a glaucoma therapeutic agent and contacting a biological sample from the subject with an LRRC4 family mimetic molecule described herein to measure the expression level of FAM19A5 protein. In some aspects, the therapeutic agent for glaucoma comprises a FAM19A5 antagonist, an LRRC4 family mimetic molecule, or both.

[0151] In some aspects, provided herein is a method of treating a retinopathy in a subject in need thereof, comprising administering to the subject a therapeutic agent for the retinopathy, and contacting a biological sample from the subject with an LRRC4 family mimetic molecule described herein to measure an expression level of FAM19A5 protein. In some aspects, the therapeutic agent for the retinopathy comprises a FAM19A5 antagonist, an LRRC4 family mimetic molecule, or both. In some aspects, a retinopathy that may be treated by the present invention comprises diabetic retinopathy. The term "diabetic retinopathy" includes all types of diabetic retinopathy, including, but not limited to, non-proliferative diabetic retinopathy (NPDR), proliferative diabetic retinopathy (PDR), diabetic macular disease, and diabetic macular edema. Without being bound to any one theory, in some aspects, a method of treating a retinopathy (e.g., diabetic retinopathy) comprises improving electroretinography in a subject in need thereof. In some aspects, improved electroretinograms include increased values ​​for A-wave, B-wave, and / or oscillatory potentials as compared to a control group (e.g., a matched subject not treated with a LRRC4 family mimetic molecule described herein).It will be apparent to one of skill in the art that the methods of treating retinal pathology may also be useful for treating other types of ocular disorders, including, but not limited to, treating macular degeneration and glaucoma.

[0152] In some aspects, the present invention provides a method of treating neuropathic pain in a subject in need thereof, comprising administering to the subject a therapeutic agent for neuropathic pain, and contacting a biological sample from the subject with an LRRC4 family mimetic molecule described herein and measuring the expression level of FAM19A5 protein. In some aspects, the therapeutic agent for neuropathic pain comprises a FAM19A5 antagonist, an LRRC4 family mimetic molecule, or both. In some aspects, the neuropathic pain is central neuropathic pain, i.e., pain due to injury or damage affecting any level of the CNS, including the central somatosensory nervous system (e.g., brain injury and spinal cord injury), or associated with a disease or disorder such as stroke, multiple sclerosis, or paramedullary infarction. In some aspects, the neuropathic pain is peripheral neuropathic pain, pain due to injury caused by or associated with a disease or disorder affecting any level of the peripheral nervous system (e.g., injury to motor nerves, sensory nerves, autonomic nerves, or a combination thereof).

[0153] In some aspects, a method of treating spinal cord injury in a subject in need thereof is provided, comprising administering to the subject a therapeutic agent for spinal cord injury, and contacting a biological sample from the subject with an LRRC4 family mimetic molecule described herein to measure an expression level of FAM19A5 protein. In some aspects, the therapeutic agent for spinal cord injury comprises a FAM19A5 antagonist, an LRRC4 family mimetic molecule, or both. In some aspects, a method of treating traumatic brain injury in a subject in need thereof is provided, comprising administering to the subject a therapeutic agent for traumatic brain injury, and contacting a biological sample from the subject with an LRRC4 family mimetic molecule described herein to measure an expression level of FAM19A5 protein. In some aspects, the therapeutic agent for traumatic brain injury comprises a FAM19A5 antagonist, an LRRC4 family mimetic molecule, or both. In some aspects, a method of treating stroke in a subject in need thereof is provided, comprising administering to the subject a therapeutic agent for stroke, and contacting a biological sample from the subject with an LRRC4 family mimetic molecule described herein to measure expression of FAM19A5 protein. In some aspects, the therapeutic agent for stroke comprises a FAM19A5 antagonist, an LRRC4 family mimetic molecule, or both.

[0154] In some aspects, a method of treating Parkinson's disease in a subject in need thereof is provided, comprising administering to the subject a therapeutic agent for Parkinson's disease, and contacting a biological sample from the subject with an LRRC4 family mimetic molecule described herein to measure expression of FAM19A5 protein. In some aspects, the therapeutic agent for Parkinson's disease comprises a FAM19A5 antagonist, an LRRC4 family mimetic molecule, or both. As used herein, the term "Parkinson's disease (PD)" refers to a neurodegenerative disorder that induces motor and non-motor symptoms and is characterized by widespread degeneration of dopamine neurons in the nigrostriatal system. Non-limiting examples of motor and non-motor manifestations of PD are provided elsewhere in this disclosure. Proteinopathy (abnormal aggregation of α-synuclein) is a hallmark of PD. Other exemplary features of PD include dopaminergic neuron injury, mitochondrial dysfunction, neuroinflammation, proteostasis (e.g., autophagic removal of damaged proteins and organelle glial dysfunction), and combinations thereof.

[0155] Additionally, any of the therapeutic agents described herein (e.g., FAM19A5 antagonists and / or LRRC4 family mimetic molecules) may be administered to a subject with one or more additional therapeutic agents. Any therapeutic agent useful for treating a disease or disorder associated with increased FAM19A5 protein expression may be used with the therapeutic agents described herein. In some aspects, the additional therapeutic agent comprises an acetylcholinesterase inhibitor. In some aspects, the additional therapeutic agent comprises a dopaminergic agent. In some aspects, the additional therapeutic agent comprises a dopamine receptor antagonist. In some aspects, the additional therapeutic agent comprises an antipsychotic agent.

[0156] Non-limiting examples of such formulations include: tetrabenazine (XENAZINE), antipsychotic drugs such as haloperidol (HALODOL), chlorpromazine, risperidone (rRIDPERDAL), quetiapine (SEROQUEL), levodopa (with or without carbidopa) (LODOSYN), dopamine agonists such as pramipexole (MIRAPEX), ropinirole (REQUIP), and rotigotine (NEUPRO), and apromorphine (APMO). rphine (Apokyn), selegiline (ELDEPRYL, ZELAPAR), rasagiline (AZILECT), entacapone (COMTAN), benztropine (COGENTIN), trihexyphenidyl, amantadine, donepezil (ARICEPT), galantamine (RAZADYNE), rivastigmine (EXELON), glatiramer acetate acetate (COPAXONE), dimethyl fumarate (TECFIDERA), fingolimod (GILENYA), teriflunomide (AUBAGIO), natalizumab (TYSABRI), alemtuzumab (LEMTRADA), mitoxantrone (NOVANTRONE), riluzole (RILUTEK), physostigmine salicylate (ANTILIRIUM), physostigmine sulfatesulfate (ESERINE), metrifonate, neostigmine, ganstigmine, pyridostigmine (MESTINON), ambenonium (MYTELASE), demarcarium, Debio 9902 (ZT-1); Debiopharmladostigil, NP-0361, tacrine (COGNEX), tolserine, velnacrine maleate, memoquin, huperzine A (HUP-A); NeuroHitech, phenserine, edrophonium (ENLON, TENSILON), INM-176, apomorphine (APOKYN), bromocriptine (PARLODEL), cabergoline (DOSTINEX), dihydrexidine, dihydroergocryptine, fenoldopam (CORLOPAM), lisuride (DOPERGIN), terguride spergolide (PERMAX), piribedil (TRIVASTAL, TRASTAL), quinpirole, SKF-82958 (GlaxoSmithKline), cariprazine, pardoprunox, sarizotan, chlorpromazine, fluphenazineloxzpine, resperidone, thioridazine, thiothixene, trifluoperazine, 7-hydroxyamoxapine, droperidol (INAPSINE, DRIDOL, DROPLETAN), domperidone (MOTILIUM), L-741742, L-745870, raclopride, SB-277011A, SCH-23390, ecopipam, SKF-83566, metoclopramide (REGLAN), lurasidone (also known as LATUDA, SM-13496; Dainippon Sumitomo, aripiprazole (ABILIFY), chlorpromazine (THORAZINE), iloperidone (FANAPTA), flupentixol decanoate (DEPIXOL, FLUANXOL), reserpine (SERPLAN), pimozide (ORAP), fluphenazine decanoate, fluphenazine hydrochloridehydrochloride), prochlorperazine (COMPRO), asenapine (SAPHRIS), loxapine (LOXITANE), molindone (MOBAN), perphenazine, thioridazine, thiothixine, trifluoperazine (STELAZINE), ramelteon, Clozapine (CLOZARIL), norclozapine (ACP-104), paliperidone (INVEGA), melperone, olanzapine (ZYPREXA), talnetant, amisulpride, ziprasidone (GEODON), blonanserin (LONASEN), ACP-103 (Acadia Pharmaceuticals, selegiline hydrochloride (I-deprenyl, ELDEPRYL, ZELAPAR), dimethylselegilene, brofaromine, phenelzine (NARDIL), tranylcypromine (PARNATE), moclobemide (AURORIX, MANERIX), befloxatone, safinamide, isocarboxazid (MARPLAN), nialamide (NIAMID), iproniazide (MARSILID, IPROZID, IPRONID), CHF-3381 (ChiesiFarmaceutici, iproclozide, toloxatone (HUMORYL, PERENUM), bifemelane, desoxypeganine, harmine (telepathine, or banasterine), harmaline, linezolid (ZYVOX, ZYVOXID), pargyline (EUDATIN, SUPIRDYL), nitecapone, tolcapone (TASMAR), tropolone, memantine (NAMENDA, AXURA, EBIXA), amantadine (SYMMET REL), acamprosate (CAMPRAL), besonprodil, ketamine (KETALAR), delucemine, dexanabinol, dexefaroxan, dextromethorphan, dextrorphan, traxoprodil, CP-283097, himantane, idantadol, ipenoxazone, L-701252 (Merck), lancicemine, levorphanol (DROMORAN), LY-233536 and LY-235959 (bothLilly, methadone (DOLOPHINE), neramexane, perzinfotel, phencyclidine, tianeptine (STABLON), dizocilpine (MK-801), EAB-318 (Wyeth), ibogaine, voacangine, tiletamine, aptiganel (CERESOTAT), gavestenel, remacimide ), MBP-8298 (synthetic myelin basic protein peptide), roquinimex (LINOMIDE), laquinimod (ABR-215062 and SAIK-MS), ABT-874 (human anti-IL-12 antibody; Abbott), rituximab (RITUXAN), leflunomide, ciclesonide, daclizumab (ZENAPAX), methotrexate (TREXALL, RHEUMATREX), suplatast tosilate tosilate, mycophenolate mofetil (CELLCEPT), mycophenolate sodium (MYFORTIC), azathioprine (AZASAN, IMURAN), mercaptopurine (PURI-NETHOL), cyclophosphamide (NEOSAR, CYTOXAN7), voclosporin, PUR-118, AMG 357, AMG 811, BCT197, chlorambucil (LEUKERAN), cladribine (LEUSTATIN, MYLINAX), alpha-fetoprotein, etanercept (ENBREL), leflunomide, cyclo Ciclesonide chloroquine, hydroxychloroquine, d-penicillamine, auranofin, sulfasalazine, sodium gold thiomalate aurothiomalate, cyclosporine, cromolyn, infliximab, adalimumab, certolizumab, pegol, golimumab, rituximab, ocrelizumab, ofatumumab, 4-benzyloxy-5-((5-undecyl-2H-pyrrol-2-ylidene)methyl)-2,2'-bi-1H-pyrrole (PNU-156804), and combinations thereof.

[0157] In some aspects, any of the therapeutic agents described herein (e.g., FAM19A5 antagonists and / or LRRC4 family mimetic molecules) are administered intravenously, orally, parenterally, transthecally, intrathecally, intracerebroventricularly, intravitreally, pulmonary, subcutaneously, intradermally, intramuscularly, or intraventricularly.

[0158] IV. Mimetic molecules The present invention provides LRRC4 protein family member mimetic molecules ("LRRC4 family mimetic molecules") that can be used in the methods of the invention. As will be apparent from the present invention, the LRRC4 family mimetic molecules described herein share certain properties with LRRC4 protein family members, but are distinct from naturally occurring LRRC4 protein family members. For example, in some aspects, LRRC4 family mimetic molecules useful in the present invention include small molecules. As described elsewhere herein, in some aspects, the LRRC4 family mimetic molecules include a polypeptide, the polypeptide comprising, consisting of, or consisting essentially of a domain of a LRRC4 protein family member, the domain (also designated as a "FAM19A5 binding domain") being capable of binding to a FAM19A5 protein. In that aspect, the polypeptide may include one or more amino acid substitutions within the FAM19A5 binding domain. As described elsewhere herein, in some aspects, such amino acid substitutions increase one or more properties of the polypeptide, such as the stability and / or binding affinity of the polypeptide to the FAM19A5 protein. In some aspects, the polypeptide may comprise a FAM19A5 binding domain that lacks one or more other domains of the LRRC4 protein family member. For example, in some aspects, the polypeptide comprises the FAM19A5 binding domain but does not comprise a transmembrane domain. In some aspects, the polypeptide comprises the FAM19A5 binding domain but does not comprise an intracellular domain (e.g., a postsynaptic density binding (PB) domain) of the LRRC4 protein family member. In some aspects, the polypeptide comprises the FAM19A5 binding domain but does not comprise either the transmembrane domain or the intracellular domain. Thus, in some aspects, the polypeptides described herein are shorter than naturally occurring LRRC4 protein family members.Furthermore, when performing their other biological activities (e.g., neural circuit formation), members of the LRRC4 protein family (LRRC4, LRRC4B, and LRRC4C) interact with ligands (netrin-G2, receptor tyrosine phosphatase LAR, and netrin-G1), respectively (see, e.g., Li et al., Mol Cancer 13:266 (Dec. 2014)). Because the polypeptides of the present invention do not contain all domains of the LRRC4 protein family members, in some aspects, the polypeptides do not bind to LRRC4 protein family ligands, but instead specifically target FAM19A5 proteins. Thus, in some aspects, the polypeptides described herein do not replace endogenous LRRC4 protein family members. Instead, in some aspects, by inhibiting, reducing, and / or dissociating the interaction between FAM19A5 and LRRC4 protein family members, the polypeptides of the present invention can free endogenous LRRC4 family proteins to perform their native biological functions.

[0159] The LRRC4 family mimetic molecule comprises a small molecule compound, and in some aspects, the mimetic molecule comprises:

[0160] [ka]

[0161] or a pharma- ceutically acceptable salt thereof: where (i) R1, R2 and R3 are hydrogen, fluoro, chloro, bromo, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, t-butyl, n-pentyl, iso-pentyl, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 1-fluoroethyl, 2,2-difluoroethyl, 1,2-difluoroethyl, 1,1-difluoroethyl, 2,2,2-trifluoroethyl, methoxy, ethoxy, n-furopyroxy, iso-furopyroxy, n-butoxy, trifluoromethoxy, difluoromethyl, independently selected from methoxy, fluoromethoxy, acetyl, propionyl, n-butanoyl, iso-butanoyl, n-pentanoyl, nitro, amino, N-methylamino, N-ethylamino, Nn-propylamino, N,N-dimethylamino, N-acetylamino, N-propionylamino, N-(trifluoroacetyl)amino, formyl, hydroxy, methylthio, ethylthio, n-propylthio, methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, phenyl, hydroxymethyl, 1-hydroxyethyl, and 2-hydroxyethyl; (ii)

[0162] [ka]

[0163] is a single or double bond; (iii) Z is a linear or branched (C1-C8) alkyl, a linear or branched (C2-C8) alkenyl, a linear or branched (C2-C8) alkynyl, a (C3-C8) cycloalkyl, a (C5-C8) cycloalkenyl, a (3-8 membered) heterocycloalkyl, a (C7-C 14 )bicycloalkyl, (C7-C 14 )bicycloalkenyl, (7-14 membered)heterobicycloalkyl, (C6-C 10 )aryl, (5-10 membered)heteroaryl, and -CH-C(O)-CH=CH-Q, where Q is selected from (C3-C8)cycloalkyl, (C5-C8)cycloalkenyl, (3-8 membered)heterocycloalkyl, (C6-C 10)aryl, and (5-6 membered)heteroaryl; each cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl is optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from (C1-C6)alkoxy, (C1-C6)alkyl, halo, (C1-C6)haloalkoxy, nitro, amino, N-methylamino, N-ethylamino, N-N-propylamino, N,N-dimethylamino, formyl, and hydroxy; (iv) L is a single, double, or triple bond.

[0164] In some aspects, R1, R2 and R3 are selected from hydrogen, hydroxy, methoxy, ethoxy, n-furopyroxy, n-butoxy, trifluoromethoxy, difluoromethoxy and fluoromethoxy. In some aspects, R1, R2 and R3 are selected from hydrogen, hydroxy, methoxy, ethoxy, n-furopyroxy, iso-furopyroxy and n-butoxy. In some aspects, R1, R2 and R3 are selected from hydrogen, hydroxy and methoxy. In some aspects, R1 and R2 are selected from hydroxy and methoxy and R3 is hydrogen.

[0165] In some aspects, Z is selected from linear or branched (C1-C8) alkyl, linear or branched (C2-C8) alkenyl, linear or branched (C2-C8) alkynyl, and -CH-C(O)-CH=CH-Q, where Q is (C6-C 10 )aryl, and (5-6 membered)heteroaryl; said aryl and heteroaryl are optionally substituted with one, two, or three substituents independently selected from (C1-C6)alkoxy, (C1-C6)alkyl, halo, (C1-C6)haloalkoxy, and hydroxy. In some aspects, Z is selected from linear or branched (C1-C8)alkyl, linear or branched (C2-C8)alkenyl, and CH-C(O)-CH=CH-Q, where Q is a (C6-C8)alkyl optionally substituted with one, two, or three substituents independently selected from (C1-C6)alkoxy, (C1-C6)alkyl, halo, (C1-C6)haloalkoxy, and hydroxy. 10) aryl.

[0166] In some aspects, the LRRC4 family mimetic molecule is a small molecule of Formula I: wherein (i) R1, R2, and R3 are selected from hydrogen, hydroxy, methoxy, ethoxy, n-furopyroxy, isofuropyroxy, n-butoxy, trifluoromethoxy, difluoromethoxy, and fluoromethoxy; (ii)

[0167] [ka]

[0168] is a single or double bond; (iii) Z is selected from linear or branched (C1-C8) alkyl, linear or branched (C2-C8) alkenyl, linear or branched (C2-C8) alkynyl, and -CH-C(O)-CH=CH-Q, wherein Q is (C6-C 10 )aryl, and (5-6 membered)heteroaryl; said aryl and heteroaryl are optionally substituted with 1, 2, or 3 substituents independently selected from (C1-C6)alkoxy, (C1-C6)alkyl, halo, (C1-C6)haloalkoxy, and hydroxy; (iv) L is a double or triple bond.

[0169] In some aspects, the LRRC3 family mimetic molecule is a small molecule of Formula I: wherein (i) R1, R2, and R3 are selected from hydrogen, hydroxy, methoxy, ethoxy, n-furopyroxy, isofuropyroxy, and n-butoxy; (ii)

[0170] [ka]

[0171] is a single or double bond; (iii) Z is selected from linear or branched (C1-C8) alkyl, linear or branched (C2-C8) alkenyl, and -CH-C(O)-CH=CH-Q, wherein Q is a (C6-C6) alkyl group optionally substituted with one, two, or three substituents independently selected from (C1-C6) alkoxy, (C1-C6) alkyl, halo, (C1-C6) haloalkoxy, and hydroxy. 10 ) aryl; (iv) L is a double or triple bond.

[0172] In some aspects, the LRRC4 family mimetic molecule is a mimetic molecule selected from the following structural formulas:

[0173] [ka]

[0174] or a pharma- ceutically acceptable salt thereof.

[0175] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0176] [ka]

[0177] or a pharma- ceutically acceptable salt thereof.

[0178] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0179] [ka]

[0180] or a pharma- ceutically acceptable salt thereof.

[0181] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0182] [ka]

[0183] or a pharma- ceutically acceptable salt thereof.

[0184] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0185] [ka]

[0186] or a pharma- ceutically acceptable salt thereof.

[0187] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0188] [ka]

[0189] or a pharma- ceutically acceptable salt thereof: where (i) R1, R2 and R3 are hydrogen, fluoro, chloro, bromo, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, t-butyl, n-pentyl, iso-pentyl, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 1-fluoroethyl, 2,2-difluoroethyl, 1,2-difluoroethyl, 1,1-difluoroethyl, 2,2,2-trifluoroethyl, methoxy, ethoxy, n-furopyroxy, iso-furopyroxy, n-butoxy, trifluoromethoxy, difluoromethyl, independently selected from methoxy, fluoromethoxy, acetyl, propionyl, n-butanoyl, iso-butanoyl, n-pentanoyl, nitro, amino, N-methylamino, N-ethylamino, Nn-propylamino, N,N-dimethylamino, N-acetylamino, N-propionylamino, N-(trifluoroacetyl)amino, formyl, hydroxy, methylthio, ethylthio, n-propylthio, methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, phenyl, hydroxymethyl, 1-hydroxyethyl, and 2-hydroxyethyl; (ii)

[0190] [ka]

[0191] is a single or double bond; (iii) Z is a linear or branched (C1-C8) alkyl, a linear or branched (C2-C8) alkenyl, a linear or branched (C2-C8) alkynyl, a (C3-C8) cycloalkyl, a (C5-C8) cycloalkenyl, a (3-8 membered) heterocycloalkyl, a (C7-C 14 )bicycloalkyl, (C7-C 14 )bicycloalkenyl, (7-14 membered)heterobicycloalkyl, (C6-C 10 )aryl, (5-10 membered)heteroaryl, and -CH-C(O)-CH=CH-Q, where Q is selected from (C3-C8)cycloalkyl, (C5-C8)cycloalkenyl, (3-8 membered)heterocycloalkyl, (C6-C 10)aryl, and (5-6 membered)heteroaryl; each cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl is optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from (C1-C6)alkoxy, (C1-C6)alkyl, halo, (C1-C6)haloalkoxy, nitro, amino, N-methylamino, N-ethylamino, N-N-propylamino, N,N-dimethylamino, formyl, and hydroxy; (iv) L is a single, double, or triple bond; The LRRC4 family mimetic molecule is a mimetic molecule selected from the following structural formulas:

[0192] [ka]

[0193] or a pharma- ceutically acceptable salt thereof.

[0194] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0195] [ka]

[0196] or a pharma- ceutically acceptable salt thereof: where (i) R1, R2 and R3 are hydrogen, fluoro, chloro, bromo, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, t-butyl, n-pentyl, iso-pentyl, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 1-fluoroethyl, 2,2-difluoroethyl, 1,2-difluoroethyl, 1,1-difluoroethyl, 2,2,2-trifluoroethyl, methoxy, ethoxy, n-furopyroxy, iso-furopyroxy, n-butoxy, trifluoromethoxy, difluoromethyl, independently selected from methoxy, fluoromethoxy, acetyl, propionyl, n-butanoyl, iso-butanoyl, n-pentanoyl, nitro, amino, N-methylamino, N-ethylamino, Nn-propylamino, N,N-dimethylamino, N-acetylamino, N-propionylamino, N-(trifluoroacetyl)amino, formyl, hydroxy, methylthio, ethylthio, n-propylthio, methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, phenyl, hydroxymethyl, 1-hydroxyethyl, and 2-hydroxyethyl; (ii) Z is a linear or branched (C1-C8) alkyl, a linear or branched (C2-C8) alkenyl, a linear or branched (C2-C8) alkynyl, a (C3-C8) cycloalkyl, a (C5-C8) cycloalkenyl, a (3-8 membered) heterocycloalkyl, a (C7-C 14 )bicycloalkyl, (C7-C 14 )bicycloalkenyl, (7-14 membered)heterobicycloalkyl, (C6-C 10 )aryl, (5-10 membered)heteroaryl, and -CH=CH-Q, where Q is selected from (C3-C8)cycloalkyl, (C5-C8)cycloalkenyl, (3-8 membered)heterocycloalkyl, (C6-C 10)aryl, and (5-6 membered)heteroaryl; each cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl is optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from (C1-C6)alkoxy, (C1-C6)alkyl, halo, (C1-C6)haloalkoxy, nitro, amino, N-methylamino, N-ethylamino, N-N-propylamino, N,N-dimethylamino, formyl, and hydroxy; (iii) L is a single, double, or triple bond.

[0197] In some aspects, R1, R2 and R3 are selected from hydrogen, fluoro, chloro, bromo, hydroxy, methoxy, ethoxy, n-furopyroxy, iso-furopyroxy, n-butoxy, trifluoromethoxy, difluoromethoxy, fluoromethoxy, amino, N-methylamino, N-ethylamino, N-propylamino, and N,N-dimethylamino. In some aspects, R1, R2 and R3 are selected from hydrogen, fluoro, chloro, bromo, hydroxy, methoxy, ethoxy, n-furopyroxy, amino, N-methylamino, N-ethylamino, N-propylamino, and N,N-dimethylamino. In some aspects, R1, R2 and R3 are selected from hydrogen, fluoro, hydroxy, methoxy, and N,N-dimethylamino. In some aspects, R1 and R2 are selected from fluoro, hydroxy, methoxy, and N,N-dimethylamino and R3 is hydrogen.

[0198] In some aspects, Z is selected from linear or branched (C1-C8) alkyl, linear or branched (C2-C8) alkenyl, linear or branched (C2-C8) alkynyl, and -CH=CH-Q; Q is selected from (C6-C 10)aryl, and (5-6 membered)heteroaryl; said aryl and heteroaryl are optionally substituted with one, two, or three substituents independently selected from (C1-C6)alkoxy, (C1-C6)alkyl, halo, (C1-C6)haloalkoxy, and hydroxy. In some aspects, Z is selected from linear or branched (C1-C8)alkyl, linear or branched (C2-C8)alkenyl, and -CH=CH-Q; Q is (C6-C8)alkoxy, (C1-C6)alkyl, halo, (C1-C6)haloalkoxy, and hydroxy. 10 ) aryl.

[0199] In some aspects, the LRRC4 family mimetic molecule is a small molecule of formula (II): where (i) R1, R2, and R3 are selected from hydrogen, fluoro, chloro, bromo, hydroxy, methoxy, ethoxy, n-furopyroxy, iso-furopyroxy, n-butoxy, trifluoromethoxy, difluoromethoxy, fluoromethoxy, amino, N-methylamino, N-ethylamino, N-N-propylamino, and N,N-dimethylamino. In some aspects, R1, R2, and R3 are selected from hydrogen, fluoro, chloro, bromo, hydroxy, methoxy, ethoxy, n-furopyroxy, amino, N-methylamino, N-ethylamino, N-N-propylamino, and N,N-dimethylamino; (ii) Z is selected from linear or branched (C1-C8) alkyl, linear or branched (C2-C8) alkenyl, linear or branched (C2-C8) alkynyl, and -CH=CH-Q; Q is selected from (C6-C 10 )aryl, and (5-6 membered)heteroaryl; said aryl and heteroaryl are optionally substituted with 1, 2, or 3 substituents independently selected from (C1-C6)alkoxy, (C1-C6)alkyl, halo, (C1-C6)haloalkoxy, and hydroxy; (iii) L is a double bond.

[0200] In some aspects, the LRRC4 family mimetic molecule is a small molecule of formula (II): wherein (i) R1, R2, and R3 are selected from hydrogen, fluoro, chloro, bromo, hydroxy, methoxy, ethoxy, n-furopyroxy, amino, N-methylamino, N-ethylamino, N-N-propylamino, and N,N-dimethylamino; (ii) Z is selected from linear or branched (C1-C8) alkyl, linear or branched (C2-C8) alkenyl, and -CH=CH-Q; Q is (C6-C6) alkyl optionally substituted with 1, 2, or 3 substituents independently selected from (C1-C6) alkoxy, (C1-C6) alkyl, halo, (C1-C6) haloalkoxy, and hydroxy. 10 ) aryl; (iii) L is a double bond.

[0201] In some aspects, the LRRC4 family molecule is a mimetic molecule selected from the following structural formulas:

[0202] [ka]

[0203] or a pharma- ceutically acceptable salt thereof.

[0204] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0205] [ka]

[0206] or a pharma- ceutically acceptable salt thereof.

[0207] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0208] [ka]

[0209] or a pharma- ceutically acceptable salt thereof.

[0210] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0211] [ka]

[0212] or a pharma- ceutically acceptable salt thereof.

[0213] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0214] [ka]

[0215] or a pharma- ceutically acceptable salt thereof.

[0216] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0217] [ka]

[0218] or a pharma- ceutically acceptable salt thereof.

[0219] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0220] [ka]

[0221] or a pharma- ceutically acceptable salt thereof.

[0222] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0223] [ka]

[0224] or a pharma- ceutically acceptable salt thereof: where (i) R1, R2 and R3 are hydrogen, fluoro, chloro, bromo, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, t-butyl, n-pentyl, iso-pentyl, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 1-fluoroethyl, 2,2-difluoroethyl, 1,2-difluoroethyl, 1,1-difluoroethyl, 2,2,2-trifluoroethyl, methoxy, ethoxy, n-furopyroxy, iso-furopyroxy, n-butoxy, trifluoromethoxy, difluoromethyl, independently selected from methoxy, fluoromethoxy, acetyl, propionyl, n-butanoyl, iso-butanoyl, n-pentanoyl, nitro, amino, N-methylamino, N-ethylamino, Nn-propylamino, N,N-dimethylamino, N-acetylamino, N-propionylamino, N-(trifluoroacetyl)amino, formyl, hydroxy, methylthio, ethylthio, n-propylthio, methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, phenyl, hydroxymethyl, 1-hydroxyethyl, and 2-hydroxyethyl; (ii) Z is a linear or branched (C1-C8) alkyl, a linear or branched (C2-C8) alkenyl, a linear or branched (C2-C8) alkynyl, -Y-(C3-C8) cycloalkyl, -Y-(C5-C8) cycloalkenyl, -Y-(3-8 membered) heterocycloalkyl, -Y-(C7-C 14 )bicycloalkyl, -Y-(C7-C 14 )bicycloalkenyl, -Y-(7-14 membered)heterobicycloalkyl, -Y-(C6-C 10)aryl, and -Y-(5-10 membered)heteroaryl, wherein Y is a bond or a C1-C3 straight or branched alkylene, and said cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from (C1-C6)alkoxy, (C1-C6)alkyl, halo, (C1-C6)haloalkoxy, nitro, amino, N-methylamino, N-ethylamino, N-N-propylamino, N,N-dimethylamino, formyl, and hydroxy; (iii) L is a single, double, or triple bond; (iv) n is 0 or 1.

[0225] In some aspects, R1, R2 and R3 are selected from hydrogen, hydroxy, methoxy, ethoxy, n-furopyroxy, isofuropyroxy, n-butoxy, trifluoromethoxy, difluoromethoxy and fluoromethoxy. In some aspects, R1, R2 and R3 are selected from hydrogen, hydroxy, methoxy, ethoxy, n-furopyroxy, isofuropyroxy and n-butoxy. In some aspects, R1, R2 and R3 are selected from hydrogen, hydroxy and methoxy. In some aspects, R1 and R2 are selected from hydroxy and methoxy and R3 is hydrogen.

[0226] In some aspects, Z is -Y-(C3-C8)cycloalkyl, -Y-(C5-C8)cycloalkenyl, -Y-(3-8 membered)heterocycloalkyl, -Y-(C7-C 14 )bicycloalkyl, -Y-(C7-C 14 )bicycloalkenyl, -Y-(7-14 membered)heterobicycloalkyl, -Y-(C6-C 10)aryl, and -Y-(5-10 membered)heteroaryl, where Y is a bond or a C1-C3 straight or branched alkylene, and the cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from (C1-C6)alkoxy, (C1-C6)alkyl, halo, (C1-C6)haloalkoxy, nitro, amino, N-methylamino, N-ethylamino, N-N-propylamino, N,N-dimethylamino, formyl, and hydroxy. In some aspects, Z is -Y-(C6-C 10 )aryl, and -Y-(5-10 membered)heteroaryl, where Y is a bond or a C1-C3 straight or branched alkylene, and the aryl and heteroaryl are optionally substituted with 1, 2, or 3 substituents independently selected from (C1-C6)alkoxy and halo.

[0227] In some aspects, the LRRC4 family mimetic molecule is a small molecule of formula (III): wherein (i) R1, R2, and R3 are selected from hydrogen, hydroxy, methoxy, ethoxy, n-furopyroxy, iso-furopyroxy, n-butoxy, trifluoromethoxy, difluoromethoxy, and fluoromethoxy; (ii) Z is -Y-(C3-C8)cycloalkyl, -Y-(C5-C8)cycloalkenyl, -Y-(3-8 membered)heterocycloalkyl, -Y-(C7-C 14 )bicycloalkyl, -Y-(C7-C 14 )bicycloalkenyl, -Y-(7-14 membered)heterobicycloalkyl, -Y-(C6-C 10)aryl, and -Y-(5-10 membered)heteroaryl, wherein Y is a bond or a C1-C3 straight or branched alkylene, and said cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from (C1-C6)alkoxy, (C1-C6)alkyl, halo, (C1-C6)haloalkoxy, nitro, amino, N-methylamino, N-ethylamino, N-N-propylamino, N,N-dimethylamino, formyl, and hydroxy; (iii) L is a triple bond.

[0228] In some aspects, the LRRC4 family mimetic molecule is a small molecule of formula (III): wherein (i) R1, R2, and R3 are selected from hydrogen, hydroxy, methoxy, ethoxy, n-furopyroxy, iso-furopyroxy, and n-butoxy; (ii) Z is -Y-(C6-C 10 )aryl, and -Y-(5-10 membered)heteroaryl, wherein Y is a bond or a C1-C3 straight or branched alkylene, said aryl and heteroaryl are optionally substituted with 1, 2, or 3 substituents independently selected from (C1-C6)alkoxy and halo; (iii) L is a triple bond.

[0229] In some aspects, the LRRC4 family mimetic molecule is a mimetic molecule selected from the following structural formulas:

[0230] [ka]

[0231] or a pharma- ceutically acceptable salt thereof.

[0232] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0233] [ka]

[0234] or a pharma- ceutically acceptable salt thereof.

[0235] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0236] [ka]

[0237] or a pharma- ceutically acceptable salt thereof.

[0238] In some aspects, the LRRC4 family mimetic molecule has the following structural formula:

[0239] [ka]

[0240] As described herein, in some aspects, the LRRC4 family mimetic molecules useful in the present invention comprise a polypeptide, said polypeptide comprising at least a FAM19A5-binding domain of a LRRC4 protein family member. Unless otherwise indicated, the total length of the FAM19A5-binding domain is not particularly limited, provided that the domain is capable of binding to the FAM19A5 protein. In some aspects, the FAM19A5 binding domain is at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, at least about 20, at least about 21, at least about 22, at least about 23, at least about 24, at least about 25, at least about 26, at least about 27, at least about 28, at least about 29, or at least about 30 amino acids in length. In some aspects, the FAM19A5 binding domain is about 10 to about 23 amino acids in length. In some aspects, the FAM19A5 binding domain is about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, or about 23 amino acids in length. In some aspects, the FAM19A5 binding domain of the LRRC4 family mimetic molecule is about 10 amino acids in length.

[0241] In some aspects, a polypeptide of a LRRC4 family mimetic molecule comprises an amino acid sequence (N-terminus to C-terminus) having the following chemical formula: A-(T / S)-B (Formula IV) (SEQ ID NO: 25): where (i) "A" includes X1-(T / S)-(Y / F)-F-X5, and (ii) "B" includes (V / I)-TV-(E / V), X1 is tyrosine (Y), phenylalanine (F), valine (V), leucine (L), or isoleucine (I); (T / S) is threonine (T) or serine (S); (Y / F) is tyrosine (Y) or phenylalanine (F); X5 is any amino acid; (V / I) is valine (V) or isoleucine (I); (E / V) is glutamic acid (E) or valine (V).

[0242] In some aspects, the polypeptide of the LRRC4 family mimetic molecules described herein comprises an amino acid sequence (N-terminus to C-terminus) having the following chemical formula: A-(T / S)-B (Formula IV) (SEQ ID NO: 26): where (i) "A" includes (Y / W / M)-(T / Y)-(Y / W)-(F / Y / W)-(T / Y), and (ii) "B" includes X7-(T / S / Y)-X9-X10; (Y / W / M) is tyrosine (Y), tryptophan (W), or methionine (M); (T / Y) is threonine (T) or tyrosine (Y); (Y / W) is tyrosine (Y) or tryptophan (W); (F / Y / W) is phenylalanine (F), tyrosine (Y), or tryptophan (W); X7 is valine (V), tyrosine (Y), phenylalanine (F), leucine (L), tryptophan (W), or methionine (M); (T / S / Y) is threonine (T), serine (S), or tyrosine (Y); X9 is valine (V), isoleucine (I), tyrosine (Y), phenylalanine (F), leucine (L), tryptophan (W), or methionine (M); X10 is glutamic acid (E), aspartic acid (D), isoleucine (I), tyrosine (Y), phenylalanine (F), methionine (M), or tryptophan (W).

[0243] In some aspects, LRRC4 family mimetic molecules useful in the present invention include polypeptides comprising an amino acid sequence (N-terminus to C-terminus) having the following chemical formula: X1-X2-X3-F-X5-T-X7-TV-X10 (Formula V) (SEQ ID NO: 27): where X1 is Y, F, V, L, or I; X2 is T or S; X3 is Y or F; X5 is any amino acid; X7 is V or I; and / or X10 is E or V; The polypeptide is capable of binding to a FAM19A5 protein, thereby inhibiting, reducing and / or dissociating the interaction between the FAM19A5 protein and a member of the LRRC4 protein family.

[0244] In some aspects, the LRRC4 family mimetic molecules described herein include a polypeptide comprising an amino acid sequence (N-terminus to C-terminus) of the following formula: X1-X2-X3-X4-X5-X6-X7-X8-X9-X10 (Formula VI) (SEQ ID NO: 28): where X1 is Y, F, V, L, I, W, or M; X2 is T, S, or Y; X3 is Y, F, or W; X4 is F, Y, or W; X5 is any amino acid, e.g., T, S, or Y; X6 is T, S, or Y; X7 is V, I, Y, F, L, W, or M; X8 is T, S, or Y; X9 is V, I, Y, F, L, W, or M; and / or X10 is E, D, V, I, Y, F, M, or W; The polypeptide is capable of binding to a FAM19A5 protein, thereby inhibiting, reducing and / or dissociating the interaction between the FAM19A5 protein and a member of the LRRC4 protein family.

[0245] For any of the above LRRC4 family mimetic molecules, in some aspects, (i) X1 is Y, F, V, L, or I; (ii) X2 is T or S; (iii) X3 is Y or F; (iv) X4 is F; (v) X5 is T or S; (vi) X6 is T; (vii) X7 is V or I; (viii) X8 is T; (ix) X9 is V; (x) X10 is E or V; (xi) any combination of (i)-(x). In some aspects, X1 is Y, F, V, L, or I. In some aspects, X2 is T or S. In some aspects, X3 is Y or F. In some aspects, X4 is F. In some aspects, X5 is T or S. In some aspects, X6 is T. In some aspects, X7 is V or I. In some aspects, X8 is T. In some aspects, X9 is V. In some aspects, X10 is E or V. In some aspects, the amino acid at position X2 is phosphorylated. In some aspects, the amino acid at position X2 is O-glycosylated.

[0246] In some aspects, the polypeptides of the LRRC4 family mimetic molecules described herein comprise the amino acid sequence set forth as SEQ ID NO:29 (YTYFTTVTVE) with 1, 2, 3, 4, 5, or 6 amino acids that differ (e.g., substituted) from each other in the amino acid sequence. In some aspects, the polypeptides of the LRRC4 family mimetic molecules described herein consist of the amino acid sequence set forth as SEQ ID NO:29 (YTYFTTVTVE) with 1, 2, 3, 4, 5, or 6 amino acids that differ (e.g., substituted) from each other in the amino acid sequence. In some aspects, the polypeptides of the LRRC4 family mimetic molecules described herein consist essentially of the amino acid sequence set forth as SEQ ID NO:29 (YTYFTTVTVE) with 1, 2, 3, 4, 5, or 6 amino acids that differ (e.g., substituted) from each other in the amino acid sequence.

[0247] In some aspects, the LRRC4 family mimetic molecule polypeptide comprises the amino acid sequence set forth in SEQ ID NO:29 (YTYFTTVTVE). In some aspects, the LRRC4 family mimetic molecule polypeptide consists of the amino acid sequence set forth in SEQ ID NO:29 (YTYFTTVTVE). In some aspects, the LRRC4 family mimetic molecule polypeptide consists essentially of the amino acid sequence set forth in SEQ ID NO:29 (YTYFTTVTVE). As determined herein, the amino acid sequence set forth in SEQ ID NO:29 (YTYFTTVTVE) corresponds to the FAM19A5 binding domain of the LRRC4B protein.

[0248] In some aspects, the LRRC4 family mimetic molecule polypeptide comprises the amino acid sequence set forth in SEQ ID NO:30 (YSFFTTVTVE). In some aspects, the LRRC4 family mimetic molecule polypeptide consists of the amino acid sequence set forth in SEQ ID NO:30 (YSFFTTVTVE). In some aspects, the LRRC4 family mimetic molecule polypeptide consists essentially of the amino acid sequence set forth in SEQ ID NO:30 (YSFFTTVTVE). As determined herein, the amino acid sequence set forth in SEQ ID NO:30 (YSFFTTVTVE) corresponds to the FAM19A5 binding domain of the LRRC4 protein.

[0249] In some aspects, the LRRC4 family mimetic molecule polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 31 (FSYFSTVTVE). In some aspects, the LRRC4 family mimetic molecule polypeptide consists of the amino acid sequence set forth in SEQ ID NO: 31 (FSYFSTVTVE). In some aspects, the LRRC4 family mimetic molecule polypeptide consists essentially of the amino acid sequence set forth in SEQ ID NO: 31 (FSYFSTVTVE). As determined herein, the amino acid sequence set forth in SEQ ID NO: 31 (FSYFSTVTVE) corresponds to the FAM19A5 binding domain of the LRRC4C protein.

[0250] As described herein, the FAM19A5 binding domain of LRRC4 protein family members is largely conserved among vertebrates (see FIG. 20). Thus, without being bound by any one theory, one or more amino acid residues of the amino acid sequence set forth as any one of SEQ ID NOs: 29 (YTYFTTVTVE), 30 (YSFFTTVTVE) and 31 (FSYFSTVTVE) may be substituted with amino acids present at those residues in other vertebrates. Examples of such substitutions are provided herein (see FIG. 20).

[0251] In some aspects, one or more amino acid residues of the amino acid sequence set forth in any one of SEQ ID NOs: 29 (YTYFTTVTVE), 30 (YSFFTTVTVE), and 31 (FSYFSTVTVE) may be substituted with an amino acid that shares similar biochemical properties. For example, in the amino acid sequence set forth in SEQ ID NO: 29 (YTYFTTVTVE), the Y at position 1 may be substituted with another hydrophobic amino acid (e.g., F, V, L, I, W, or M). In some aspects, the T at position 2 may be substituted with another amino acid having a similar hydroxyl group (OH) in its side chain (e.g., S or Y). In some aspects, the Y at position 3 may be substituted with another amino acid having a common aromatic ring in its side chain that can participate in van der Waals interactions (e.g., F or W). In some aspects, the F at position 4 may be substituted with an amino acid such as Y or W. In some aspects, the T at position 5 may be substituted with an amino acid such as S or Y. In some aspects, the T at position 6 may be substituted with an amino acid such as S or Y. In some aspects, the V at position 7 may be substituted with another amino acid having a side chain with a large hydrophobic volume (e.g., I, Y, F, L, W, or M). In some aspects, the T at position 8 may be substituted with another amino acid such as S or Y. In some aspects, the V at position 9 may be substituted with I, Y, F, L, W, or M. In some aspects, the E at position 10 may be substituted with another amino acid having an acidic side chain (e.g., I, Y, F, M, or W).

[0252] In some aspects, the polypeptides of the LRRC4 family mimetic molecules described herein comprise an amino acid sequence that is at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% identical to the amino acid sequence set forth in SEQ ID NO:29 (YTYFTTVTVE), and the polypeptides are capable of binding to FAM19A5 protein, thereby inhibiting, reducing, and / or dissociating the interaction between the FAM19A5 protein and the LRRC4 family of proteins. In some aspects, a polypeptide of a LRRC4 family mimetic molecule comprises an amino acid sequence that is at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence set forth in SEQ ID NO:5, and the polypeptide is capable of binding to a FAM19A5 protein, thereby inhibiting, reducing, and / or dissociating the interaction between the FAM19A5 protein and a LRRC4 protein family member. In some aspects, a polypeptide of a LRRC4 family mimetic molecule comprises an amino acid sequence that is at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence set forth in SEQ ID NO:4, and the polypeptide is capable of binding to a FAM19A5 protein, thereby inhibiting, reducing, and / or dissociating the interaction between the FAM19A5 protein and a LRRC4 protein family member.In some aspects, a polypeptide of a LRRC4 family mimetic molecule comprises an amino acid sequence that is at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence set forth in SEQ ID NO:6, and the polypeptide is capable of binding to a FAM19A5 protein, thereby inhibiting, reducing, and / or dissociating the interaction between a FAM19A5 protein and a LRRC4 protein family member.

[0253] As will be appreciated from the present invention, in some aspects, the polypeptides of the LRRC4 family mimetic molecules described herein (e.g., comprising a FAM19A5-binding domain of an LRRC4 protein family member) comprise one or more amino acid modifications. In some aspects, the one or more amino acid modifications can increase the binding affinity of the LRRC4 family mimetic molecule to the FAM19A5 protein. Thus, in some aspects, the binding affinity of the LRRC4 family mimetic molecules described herein for the FAM19A5 protein is increased by at least about 0.5-fold, at least about 1-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold, as compared to a control (e.g., a LRRC4 family mimetic molecule lacking the amino acid modification or corresponding to a naturally occurring LRRC4 protein family member). In some aspects, one or more amino acid modifications can increase the stability of the LRRC4 family mimetic molecule. Thus, in some aspects, the stability of the LRRC4 family mimetic molecules described herein is increased by at least about 0.5-fold, at least about 1-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold as compared to a control (e.g., a LRRC4 family mimetic molecule lacking an amino acid modification or a naturally occurring LRRC4 protein family mimetic molecule).

[0254] In some aspects, one or more amino acid modifications can improve the activity of the LRRC4 family mimetic molecules described herein to inhibit the interaction between the FAM19A5 protein and a LRRC4 protein family member (e.g., by increasing binding affinity and / or stability). Thus, in some aspects, the activity of the LRRC4 family mimetic molecule to inhibit the interaction between the FAM19A5 protein and a LRRC4 protein family member is increased by at least about 0.5-fold, at least about 1-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, or at least about 50-fold compared to a control (e.g., a LRRC4 mimetic molecule lacking the amino acid modifications or a naturally occurring LRRC4 protein family mimetic molecule).

[0255] Non-limiting examples of amino acid modifications useful in the present invention are provided throughout the present invention. For example, in some aspects, the LRRC4 family mimetic molecules described herein comprise one of the FAM19A5 binding domains of the LRRC4 protein family members, i.e., YTYFTTVTVE (SEQ ID NO: 29), YSFFTTVTVE (SEQ ID NO: 30), or FSYFSTVTVE (SEQ ID NO: 31), and one or more amino acids at the N-terminus, C-terminus, or both the N-terminus and C-terminus of the polypeptide. In some aspects, the polypeptides useful in the present invention comprise at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, or at least about 20 additional amino acids at the N-terminus of the polypeptide. In some aspects, the polypeptide comprises at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, or at least about 20 additional amino acids at the C-terminus of the polypeptide.In some aspects, the polypeptide comprises: (i) at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, or at least about 20 additional amino acids at the N-terminus of the polypeptide; and (ii) at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, or at least about 20 additional amino acids at the C-terminus of the polypeptide. As identified herein (see Example 9), in some aspects, one or more amino acids differ from the amino acid present at a particular residue in a naturally occurring LRRC4 protein family member.

[0256] For example, in some aspects, a polypeptide described herein comprises an amino acid sequence as presented as SEQ ID NO: 18 (GYTYFTTVTVETLETQPGEE) with one or more amino acid modifications (e.g., substitutions). In some aspects, a polypeptide described herein comprises an amino acid sequence as presented as SEQ ID NO: 18 (GYTYFTTVTVETLETQPGEE) with two amino acid modifications (e.g., substitutions). In some aspects, the amino acid modifications are at residues T12 and L13 of SEQ ID NO: 18. In some aspects, a polypeptide described herein comprises an amino acid sequence as presented as SEQ ID NO: 18 (GYTYFTTVTVETLETQPGEE) with one or more amino acid modifications (e.g., substitutions). In some aspects, a polypeptide described herein comprises an amino acid sequence as presented as SEQ ID NO: 18 (GYTYFTTVTVETLETQPGEE) with two amino acid modifications (e.g., substitutions). In some aspects, the amino acid modifications are at residues T12 and L13 of SEQ ID NO: 18. In some aspects, the polypeptides described herein consist essentially of the amino acid sequence set forth as SEQ ID NO: 18 (GYTYFTTVTVETLETQPGEE) with one or more amino acid modifications (e.g., substitutions). In some aspects, the polypeptides described herein consist essentially of the amino acid sequence set forth as SEQ ID NO: 18 (GYTYFTTVTVETLETQPGEE) with two amino acid modifications (e.g., substitutions). In some aspects, the amino acid modifications are at residues T12 and L13 of SEQ ID NO: 18.

[0257] In some aspects, the polypeptides described herein comprise the amino acid sequence presented as SEQ ID NO: 17 (GYTYFTTVTVETLETQ) with one or more amino acid modifications (e.g., substitutions). In some aspects, the polypeptides described herein comprise the amino acid sequence presented as SEQ ID NO: 17 (GYTYFTTVTVETLETQ) with two amino acid modifications (e.g., substitutions). In some aspects, the amino acid modifications are at residues T12 and L13 of SEQ ID NO: 17. In some aspects, the polypeptides described herein comprise the amino acid sequence presented as SEQ ID NO: 17 (GYTYFTTVTVETLETQ) with one or more amino acid modifications (e.g., substitutions). In some aspects, the polypeptides described herein comprise the amino acid sequence presented as SEQ ID NO: 17 (GYTYFTTVTVETLETQ) with two amino acid modifications (e.g., substitutions). In some aspects, the amino acid modifications are at residues T12 and L13 of SEQ ID NO: 17. In some aspects, the polypeptides described herein consist essentially of the amino acid sequence set forth as SEQ ID NO: 17 (GYTYFTTVTVETLETQ) with one or more amino acid modifications (e.g., substitutions). In some aspects, the polypeptides described herein consist essentially of the amino acid sequence set forth as SEQ ID NO: 17 (GYTYFTTVTVETLETQ) with two amino acid modifications (e.g., substitutions). In some aspects, the amino acid modifications are at residues T12 and L13 of SEQ ID NO: 17.

[0258] In some aspects, the polypeptides described herein comprise the amino acid sequence presented as SEQ ID NO: 19 (GYTYFTTVTVETLETQPGEKEPPGPTTD) with one or more amino acid modifications (e.g., substitutions). In some aspects, the polypeptides described herein comprise the amino acid sequence presented as SEQ ID NO: 19 (GYTYFTTVTVETLETQPGEKEPPGPTTD) with two amino acid modifications (e.g., substitutions). In some aspects, the amino acid modifications are at residues T12 and L13 of SEQ ID NO: 19. In some aspects, the polypeptides described herein comprise the amino acid sequence presented as SEQ ID NO: 19 (GYTYFTTVTVETLETQPGEKEPPGPTTD) with one or more amino acid modifications (e.g., substitutions). In some aspects, the polypeptides described herein comprise the amino acid sequence presented as SEQ ID NO: 19 (GYTYFTTVTVETLETQPGEKEPPGPTTD) with two amino acid modifications (e.g., substitutions). In some aspects, the amino acid modifications are at residues T12 and L13 of SEQ ID NO: 19. In some aspects, the polypeptides described herein consist essentially of the amino acid sequence set forth as SEQ ID NO: 19 (GYTYFTTVTVETLETQPGEKEPPGPTTD) with one or more amino acid modifications (e.g., substitutions). In some aspects, the polypeptides described herein consist essentially of the amino acid sequence set forth as SEQ ID NO: 19 (GYTYFTTVTVETLETQPGEKEPPGPTTD) with two amino acid modifications (e.g., substitutions). In some aspects, the amino acid modifications are at residues T12 and L13 of SEQ ID NO: 19.

[0259] In some aspects, the polypeptides described herein comprise the amino acid sequence presented as SEQ ID NO: 143 (GYTYFTTVTVETLETQPGEEA) with one or more amino acid modifications (e.g., substitutions). In some aspects, the polypeptides described herein comprise the amino acid sequence presented as SEQ ID NO: 143 (GYTYFTTVTVETLETQPGEEA) with two amino acid modifications (e.g., substitutions). In some aspects, the amino acid modifications are at residues T12 and L13 of SEQ ID NO: 143. In some aspects, the polypeptides described herein comprise the amino acid sequence presented as SEQ ID NO: 143 (GYTYFTTVTVETLETQPGEEA) with one or more amino acid modifications (e.g., substitutions). In some aspects, the polypeptides described herein comprise the amino acid sequence presented as SEQ ID NO: 143 (GYTYFTTVTVETLETQPGEEA) with two amino acid modifications (e.g., substitutions). In some aspects, the amino acid modifications are at residues T12 and L13 of SEQ ID NO: 143. In some aspects, the polypeptides described herein consist essentially of the amino acid sequence set forth as SEQ ID NO: 143 (GYTYFTTVTVETLETQPGEEA) with one or more amino acid modifications (e.g., substitutions). In some aspects, the polypeptides described herein consist essentially of the amino acid sequence set forth as SEQ ID NO: 143 (GYTYFTTVTVETLETQPGEEA) with two amino acid modifications (e.g., substitutions). In some aspects, the amino acid modifications are at residues T12 and L13 of SEQ ID NO: 143.

[0260] In some aspects, a polypeptide described herein (e.g., comprising a FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence GYTYFTTVTVEPYETQPGEE (SEQ ID NO: 123). In some aspects, a polypeptide described herein (e.g., comprising a FAM19A5 binding domain of LRRC4B) consists of the amino acid sequence GYTYFTTVTVEPYETQPGEE (SEQ ID NO: 123). In some aspects, a polypeptide described herein (e.g., comprising a FAM19A5 binding domain of LRRC4B) consists essentially of the amino acid sequence GYTYFTTVTVEPYETQPGEE (SEQ ID NO: 123).

[0261] In some aspects, a polypeptide described herein (e.g., comprising a FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence GYTYFTTVTVEMRETQPGEE (SEQ ID NO: 124). In some aspects, a polypeptide described herein (e.g., comprising a FAM19A5 binding domain of LRRC4B) consists of the amino acid sequence GYTYFTTVTVEMRETQPGEE (SEQ ID NO: 124). In some aspects, a polypeptide described herein (e.g., comprising a FAM19A5 binding domain of LRRC4B) consists essentially of the amino acid sequence GYTYFTTVTVEMRETQPGEE (SEQ ID NO: 124).

[0262] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0263] [ka]

[0264] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0265] [ka]

[0266] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0267] [ka]

[0268] It is mandatory to configure

[0269] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0270] [ka]

[0271] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0272] [ka]

[0273] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0274] [ka]

[0275] It is mandatory to configure

[0276] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0277] [ka]

[0278] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0279] [ka]

[0280] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0281] [ka]

[0282] It is mandatory to configure

[0283] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0284] [ka]

[0285] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0286] [ka]

[0287] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0288] [ka]

[0289] It is mandatory to configure

[0290] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0291] [ka]

[0292] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0293] [ka]

[0294] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0295] [ka]

[0296] It is mandatory to configure

[0297] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0298] [ka]

[0299] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0300] [ka]

[0301] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0302] [ka]

[0303] It is mandatory to configure

[0304] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0305] [ka]

[0306] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0307] [ka]

[0308] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0309] [ka]

[0310] It is mandatory to configure

[0311] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0312] [ka]

[0313] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0314] [ka]

[0315] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0316] [ka]

[0317] It is mandatory to configure

[0318] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0319] [ka]

[0320] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0321] [ka]

[0322] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0323] [ka]

[0324] It is mandatory to configure

[0325] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0326] [ka]

[0327] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0328] [ka]

[0329] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0330] [ka]

[0331] It is mandatory to configure

[0332] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0333] [ka]

[0334] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0335] [ka]

[0336] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0337] [ka]

[0338] It is mandatory to configure

[0339] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0340] [ka]

[0341] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0342] [ka]

[0343] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0344] [ka]

[0345] It is mandatory to configure

[0346] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0347] [ka]

[0348] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0349] [ka]

[0350] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0351] [ka]

[0352] It is mandatory to configure

[0353] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0354] [ka]

[0355] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0356] [ka]

[0357] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0358] [ka]

[0359] It is mandatory to configure

[0360] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0361] [ka]

[0362] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0363] [ka]

[0364] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0365] [ka]

[0366] It is mandatory to configure

[0367] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0368] [ka]

[0369] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0370] [ka]

[0371] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0372] [ka]

[0373] It is mandatory to configure

[0374] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0375] [ka]

[0376] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0377] [ka]

[0378] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0379] [ka]

[0380] It is mandatory to configure

[0381] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0382] [ka]

[0383] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0384] [ka]

[0385] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0386] [ka]

[0387] It is mandatory to configure

[0388] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0389] [ka]

[0390] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0391] [ka]

[0392] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0393] [ka]

[0394] It is mandatory to configure

[0395] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0396] [ka]

[0397] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0398] [ka]

[0399] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0400] [ka]

[0401] It is mandatory to configure

[0402] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0403] [ka]

[0404] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0405] [ka]

[0406] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0407] [ka]

[0408] It is mandatory to configure

[0409] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0410] [ka]

[0411] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0412] [ka]

[0413] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0414] [ka]

[0415] It is mandatory to configure

[0416] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0417] [ka]

[0418] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0419] [ka]

[0420] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0421] [ka]

[0422] It is mandatory to configure

[0423] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) has the amino acid sequence

[0424] [ka]

[0425] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0426] [ka]

[0427] In some aspects, a polypeptide described herein (e.g., comprising the FAM19A5 binding domain of LRRC4B) comprises the amino acid sequence

[0428] [ka]

[0429] It is mandatory to configure

[0430] In some aspects, the LRRC4 family mimetic molecules described herein comprise one or more elements capable of increasing the activity of the polypeptide to inhibit the interaction between the FAM19A5 protein and a member of the LRRC4 protein family. For example, in some aspects, the molecule comprises (i) any of the polypeptides described herein, and (ii) one or more additional amino acids at the N-terminus of the polypeptide, the C-terminus of the polypeptide, or both the N-terminus and C-terminus of the polypeptide. In some aspects, molecules useful in the invention comprise at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, or at least about 20 additional amino acids at the N-terminus of the polypeptide. In some aspects, the molecule comprises at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, or at least about 20 additional amino acids at the C-terminus of the polypeptide.In some aspects, the molecule comprises: (i) at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, or at least about 20 additional amino acids at the N-terminus of the polypeptide; and (ii) at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, or at least about 20 additional amino acids at the C-terminus of the polypeptide.

[0431] In some aspects, the LRRC4 family mimetic molecule comprises (i) a polypeptide having an amino acid sequence designated SEQ ID NO:29 (YTYFTTVTVE), and (ii) at least one additional amino acid at the N-terminus of the polypeptide. In some aspects, the LRRC4 family mimetic molecule comprises (i) a polypeptide having an amino acid sequence designated SEQ ID NO:29 (YTYFTTVTVE), and (ii) at least one additional amino acid at the C-terminus of the polypeptide. In some aspects, the LRRC4 family mimetic molecule comprises (i) a polypeptide having an amino acid sequence designated SEQ ID NO:29 (YTYFTTVTVE), and (ii) at least one additional amino acid at both the N-terminus and the C-terminus. In some aspects, the LRRC4 family mimetic molecule useful in the present invention comprises the amino acid sequence designated SEQ ID NO:18 (GYTYFTTVTVETLETQPGEE). In some aspects, the LRRC4 family mimetic molecule consists of the amino acid sequence designated SEQ ID NO:18 (GYTYFTTVTVETLETQPGEE). In some aspects, the LRRC4 family mimetic molecule consists essentially of the amino acid sequence provided as SEQ ID NO: 18 (GYTYFTTVTVETLETQPGEE). In some aspects, the LRRC4 family mimetic molecule useful in the invention comprises the amino acid sequence provided as SEQ ID NO: 17 (GYTYFTTVTVETLETQ). In some aspects, the LRRC4 family mimetic molecule consists essentially of the amino acid sequence provided as SEQ ID NO: 17 (GYTYFTTVTVETLETQ). In some aspects, the LRRC4 family mimetic molecule consists essentially of the amino acid sequence provided as SEQ ID NO: 17 (GYTYFTTVTVETLETQ). In some aspects, the LRRC4 family mimetic molecule useful in the invention comprises the amino acid sequence provided as SEQ ID NO: 19 (GYTYFTTVTVETLETQPGEKEPPGPTTD). In some aspects, the LRRC4 family mimetic molecule is comprised of the amino acid sequence set forth in SEQ ID NO: 19 (GYTYFTTVTVETLETQPGEKEPPGPTTD).In some aspects, the LRRC4 family mimetic molecule consists essentially of the amino acid sequence set forth in SEQ ID NO: 19 (GYTYFTTVTVETLETQPGEKEPPGPTTD).

[0432] In some aspects, a LRRC4 family mimetic molecule useful in the invention comprises (i) a polypeptide having an amino acid sequence provided as SEQ ID NO:30 (YSFFTTVTVE), and (ii) at least one additional amino acid at the N-terminus of the polypeptide. In some aspects, a LRRC4 family mimetic molecule comprises (i) a polypeptide having an amino acid sequence provided as SEQ ID NO:30 (YSFFTTVTVE), and (ii) at least one additional amino acid at the C-terminus of the polypeptide. In some aspects, a LRRC4 family mimetic molecule comprises (i) a polypeptide having an amino acid sequence provided as SEQ ID NO:30 (YSFFTTVTVE), and (ii) at least one additional amino acid at both the N-terminus and the C-terminus. In some aspects, a LRRC4 family mimetic molecule useful in the invention comprises the amino acid sequence provided as SEQ ID NO:20 (NYSFFTTVTVETTEISPEDTTRK). In some aspects, a LRRC4 family mimetic molecule consists of the amino acid sequence provided as SEQ ID NO:20 (NYSFFTTVTVETTEISPEDTTRK). In some aspects, the LRRC4 family mimetic molecule consists essentially of the amino acid sequence set forth in SEQ ID NO: 20 (NYSFFTTVTVETTEISPEDTTRK).

[0433] In some aspects, a LRRC4 family mimetic molecule useful in the invention comprises (i) a polypeptide having an amino acid sequence designated as SEQ ID NO: 31 (FSYFSTVTVE), and (ii) at least one additional amino acid at the N-terminus of the polypeptide. In some aspects, a LRRC4 family mimetic molecule comprises (i) a polypeptide having an amino acid sequence designated as SEQ ID NO: 31 (FSYFSTVTVE), and (ii) at least one additional amino acid at the C-terminus of the polypeptide. In some aspects, a LRRC4 family mimetic molecule comprises (i) a polypeptide having an amino acid sequence designated as SEQ ID NO: 31 (FSYFSTVTVE), and (ii) at least one additional amino acid at both the N-terminus and the C-terminus. In some aspects, a LRRC4 family mimetic molecule useful in the invention comprises the amino acid sequence designated as SEQ ID NO: 21 (NFSYFSTVTVETMEPSQDERTTR). In some aspects, a LRRC4 family mimetic molecule consists of the amino acid sequence designated as SEQ ID NO: 21 (NFSYFSTVTVETMEPSQDERTTR). In some aspects, the LRRC4 family mimetic molecule consists essentially of the amino acid sequence set forth in SEQ ID NO: 21 (NFSYFSTVTVETMEPSQDERTTR).

[0434] In some aspects, a polypeptide of a LRRC4 family mimetic molecule comprises an amino acid sequence that is at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% identical to the amino acid sequence set forth in SEQ ID NO:29 (YTYFTTVTVE), which is capable of binding to a FAM19A5 protein, and further comprises one or more hydrophobic amino acids at a terminus. In some aspects, the hydrophobic amino acids include at least 2 amino acids, at least 3 amino acids, at least 4 amino acids, at least 5 amino acids, at least 6 amino acids, at least 7 amino acids, at least 8 amino acids, at least 9 amino acids, at least 10 amino acids, at least 15 amino acids, at least 20 amino acids, at least 25 amino acids, at least 30 amino acids, at least 35 amino acids, at least 40 amino acids, at least 45 amino acids, or at least 50 amino acids at the N-terminus.

[0435] In some aspects, a polypeptide of a LRRC4 family mimetic molecule comprises an amino acid sequence that is at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% identical to the amino acid sequence set forth in SEQ ID NO:29 (YTYFTTVTVE), which is capable of binding to a FAM19A5 protein, and which further comprises one or more amino acids at the N-terminus and / or C-terminus. In some aspects, the one or more amino acids linked to the N-terminus and / or C-terminus comprise one or more amino acid sequences derived from a LRRC4B protein. In some aspects, the one or more amino acids linked to the N-terminus include at least 2 amino acids, at least 3 amino acids, at least 4 amino acids, at least 5 amino acids, at least 6 amino acids, at least 7 amino acids, at least 8 amino acids, at least 10 amino acids, at least 15 amino acids, at least 20 amino acids, at least 25 amino acids, at least 30 amino acids, at least 35 amino acids, at least 40 amino acids, at least 45 amino acids, or at least 50 amino acids at the N-terminus. In some aspects, the one or more amino acids linked to the C-terminus include at least 2 amino acids, at least 3 amino acids, at least 4 amino acids, at least 5 amino acids, at least 6 amino acids, at least 7 amino acids, at least 8 amino acids, at least 10 amino acids, at least 15 amino acids, at least 20 amino acids, at least 25 amino acids, at least 30 amino acids, at least 35 amino acids, at least 40 amino acids, at least 45 amino acids, or at least 50 amino acids at the C-terminus. In some aspects, the one or more amino acids linked to the N-terminus and / or C-terminus are linked with a linker. In some aspects, the linker is a peptide linker.

[0436] In some aspects, the one or more additional amino acids added to the N-terminus and / or C-terminus may comprise any suitable amino acid known in the art. In some aspects, the one or more additional amino acids are hydrophilic amino acids. In some aspects, the one or more additional amino acids may comprise D-amino acids. Without being bound to any one theory, in some aspects, adding one or more D-amino acids to the N-terminus and / or C-terminus of a polypeptide can improve the persistence of an LRRC4 family mimetic molecule, for example, when administered to a subject. For example, the inclusion of D-amino acids can protect the polypeptide from protease and peptidase degradation in the blood of a subject. Thus, in some aspects, a polypeptide useful in the present invention may comprise both D-amino acids and L-amino acids. For example, in some aspects, a polypeptide described herein comprises a D-amino acid at the N-terminus and L-amino acids at all other amino acid residues. In some aspects, a polypeptide described herein comprises a D-amino acid at the C-terminus and L-amino acids at all other amino acid residues. In some aspects, the polypeptides described herein contain D-amino acids at both the N-terminus and C-terminus, and L-amino acids at all other amino acid residues.

[0437] As described herein, in some aspects, the above-mentioned LRRC4 family mimetic molecules include polypeptides having an amino acid sequence set forth as any one of SEQ ID NOs: 29 (YTYFTTVTVE), 30 (YSFFTTVTVE), and 31 (FSYFSTVTVE), which have 1, 2, 3, 4, 5, or 6 amino acids that differ (e.g., are substituted) from said amino acid sequence.

[0438] In some aspects, the LRRC4 family mimetic molecules useful in the present invention include additional modifications at the N-terminus, C-terminus, or both the N-terminus and C-terminus of the polypeptide, which can increase the stability of the polypeptide. For example, in some aspects, the N-terminus of the polypeptide is methylated. Non-limiting examples of additional modifications that can be made at the N-terminus and / or C-terminus include Fmoc, PEGylation, acetylation, or combinations thereof. In some aspects, the polypeptide may be cyclized to increase stability. Such modifications can be made by any suitable method known in the art.

[0439] As described elsewhere herein, in some aspects, a molecule useful in the present invention comprises a FAM19A5 binding domain of a LRRC4 family protein member and an additional moiety, which can improve one or more properties of the molecule (e.g., the binding affinity of the molecule to the FAM19A5 protein). As demonstrated herein (see Example 10), Applicants have determined that the addition of a juxtra-membrane sequence of a LRRC4 protein family member can greatly improve the binding affinity of the molecule to the FAM19A5 protein. The juxtra-membrane sequence is highly conserved among LRRC4 family members and is provided as SEQ ID NO: 151 (LDEVMKTTK) (LRRC4 and LRRC4B) and SEQ ID NO: 152 (IDEVMKTTK) (LRRC4C) (see FIG. 22D).

[0440] Thus, in some aspects, the molecules described herein comprise the FAM19A5 binding domain of the LRRC4 protein (i.e., YSFFTTVTVE; SEQ ID NO:30) and the juxtamembrane sequence designated as SEQ ID NO:151 (LDEVMKTTK). In some aspects, the molecules described herein comprise the FAM19A5 binding domain of the LRRC4 protein (i.e., YSFFTTVTVE; SEQ ID NO:30) and the juxtamembrane sequence designated as SEQ ID NO:152 (IDEVMKTTK). In some aspects, the molecules described herein comprise the FAM19A5 binding domain of the LRRC4B protein (i.e., YTYFTTVTVE; SEQ ID NO:29) and the juxtamembrane sequence designated as SEQ ID NO:151 (LDEVMKTTK). In some aspects, the molecules described herein comprise the FAM19A5 binding domain of the LRRC4 protein (i.e., YTYFTTVTVE; SEQ ID NO:29) and the juxtamembrane sequence designated as SEQ ID NO:152 (IDEVMKTTK). In some aspects, the molecules described herein comprise the FAM19A5 binding domain of the LRRC4B protein (i.e., FSYFSTVTVE; SEQ ID NO:31) and the juxtamembrane sequence designated SEQ ID NO:151 (LDEVMKTTK). In some aspects, the molecules described herein comprise the FAM19A5 binding domain of the LRRC4 protein (i.e., FSYFSTVTVE; SEQ ID NO:31) and the juxtamembrane sequence designated SEQ ID NO:152 (IDEVMKTTK). In some aspects, the juxtamembrane is added to the C-terminus of the molecule.

[0441] As will be apparent from the present invention, any of the modifications described herein (e.g., amino acid substitutions, addition of juxtamembrane sequences, D-amino acids) to improve one or more properties of the molecule may be used in combination. For example, in some aspects, a molecule (e.g., a polypeptide) useful in the present invention comprises: (i) an amino acid sequence as set forth in SEQ ID NO: 18 (GYTYFTTVTVETLETQPGEE) with amino acid modifications at residues T12 and L13; and (ii) a juxtamembrane sequence at the C-terminus of the molecule (e.g., SEQ ID NO: 151 or SEQ ID NO: 152). In some aspects, a molecule (e.g., a polypeptide) useful in the present invention comprises: (i) an amino acid sequence as set forth in SEQ ID NO: 18 (GYTYFTTVTVETLETQPGEE) with amino acid modifications at residues T12 and L13; (ii) D-amino acids at the N-terminus and / or C-terminus; and (iii) a juxtamembrane sequence at the C-terminus of the molecule (e.g., SEQ ID NO: 151 or SEQ ID NO: 152). In some aspects, a molecule (e.g., a polypeptide) useful in the invention comprises: (i) the amino acid sequence set forth in SEQ ID NO: 17 (GYTYFTTVTVETLETQ) with amino acid modifications at residues T12 and L13; and (ii) a juxtamembrane sequence at the C-terminus of the molecule (e.g., SEQ ID NO: 151 or SEQ ID NO: 152). In some aspects, a molecule (e.g., a polypeptide) useful in the invention comprises: (i) the amino acid sequence set forth in SEQ ID NO: 17 (GYTYFTTVTVETLETQ) with amino acid modifications at residues T12 and L13; (ii) D-amino acids at the N-terminus and / or C-terminus; and (iii) a juxtamembrane sequence at the C-terminus of the molecule (e.g., SEQ ID NO: 151 or SEQ ID NO: 152). In some aspects, a molecule (e.g., a polypeptide) useful in the invention comprises: (i) the amino acid sequence set forth in SEQ ID NO: 19 (GYTYFTTVTVETLETQPGEKEPPGPTTD) with amino acid modifications at residues T12 and L13; and (ii) a juxtamembrane sequence at the C-terminus of the molecule (e.g., SEQ ID NO: 151 or SEQ ID NO: 152).In some aspects, a molecule (e.g., a polypeptide) useful in the invention comprises: (i) the amino acid sequence provided as SEQ ID NO: 19 (GYTYFTTVTVETLETQPGEKEPPGPTTD) with amino acid modifications at residues T12 and L13; (ii) D-amino acids at the N-terminus and / or C-terminus; and (iii) a juxtamembrane sequence at the C-terminus of the molecule (e.g., SEQ ID NO: 151 or SEQ ID NO: 152). In some aspects, a molecule (e.g., a polypeptide) useful in the invention comprises: (i) the amino acid sequence provided as SEQ ID NO: 143 (GYTYFTTVTVETLETQPGEEA) with amino acid modifications at residues T12 and L13; and (ii) a juxtamembrane sequence at the C-terminus of the molecule (e.g., SEQ ID NO: 151 or SEQ ID NO: 152). In some aspects, a molecule (e.g., a polypeptide) useful in the invention comprises: (i) the amino acid sequence set forth in SEQ ID NO: 143 (GYTYFTTVTVETLETQPGEEA) with amino acid modifications at residues T12 and L13; (ii) D-amino acids at the N-terminus and / or C-terminus; and (iii) a juxtamembrane sequence at the C-terminus of the molecule (e.g., SEQ ID NO: 151 or SEQ ID NO: 152).

[0442] In some aspects, the LRRC4 family mimetic molecules described herein may include one or more additional peptides that, for example, allow them to be specifically targeted to different tissues when administered to a subject. For example, in some aspects, the LRRC4 family mimetic molecules include peptides that allow the molecules to penetrate across the blood-brain barrier (also referred to herein as "BBB shuttle"). Examples of such BBB shuttles are known in the art. Non-limiting examples are provided in Table 5 (below) (see, for example, Oller-Salvia et al., Chem Soc Rev 45:4690 (2016)).

[0443] [Table 5]

[0444] The nomenclature for cyclic peptides (&) adopts the three-letter amino acid code from that described in the literature (Spengler et al., J Pept Res 65:550-555 (2005)); [Dap] stands for diaminopropionic acid.

[0445] In some aspects, the LRRC4 family mimetic molecules useful in the present invention include fusion proteins. For example, in some aspects, the LRRC4 family mimetic molecules described herein may include (i) any polypeptide of the present invention, and (ii) a half-life extending moiety. Any suitable half-life extending moiety known in the art can be used to generate the fusion proteins of the present invention. Non-limiting examples of such half-life extending moieties include: Fc, albumin, albumin binding polypeptide, Pro / Ala / Ser (PAS), C-terminal peptide of the β subunit of human chorionic gonadotropin (CTP), polyethylene glycol (PEG), long unstructured hydrophilic sequences of amino acids (XTEN), hydroxyethyl starch (HES), albumin binding small molecules, or combinations thereof.

[0446] In some aspects, the LRRC4 family mimetic molecule comprises a protein-drug conjugate. For example, in some aspects, a polypeptide of the LRRC4 family mimetic molecule may be conjugated to a therapeutic agent, such as a therapeutic agent useful for treating a disease or disorder.

[0447] The protein-drug conjugates described herein may be prepared by methods known in the art. In some aspects, the conjugation methods produce substantially (or nearly) non-immunogenic linkages, such as peptide- (i.e., amide-), sulfide-, (sterically hindered), disulfide-, hydrazone-, and ether linkages. These linkages are largely non-immunogenic and exhibit substantial stability in serum (see, e.g., Senter, PD, Curr. Opin. Chem. Biol. 13 (2009) 235-244; WO 2009 / 059278; WO 95 / 17886, the entire contents of each of which are incorporated herein by reference).

[0448] Depending on the biochemical nature of the moiety and the polypeptide, different conjugation strategies may be used (see, e.g., Hackenberger, CPR, and Schwarzer, D., Angew. Chem. Int. Ed. Engl. 47 (2008) 10030-10074). In some aspects, site-specific reactions and covalent couplings are based on modifying naturally occurring amino acids with amino acids that have reactivity orthogonal to the reactivity of other functional groups present. For example, specific cysteines in rare sequences may be enzymatically converted from aldehydes (see, Frese, MA, and Dierks, T., ChemBioChem. 10 (2009) 425-427). It is also possible to obtain specific amino acid modifications by using specific enzymatic reactivities of natural amino acids and specific enzymes in a given sequence (see, for example, Taki, M. et al., Prot. Eng. Des. Sel. 17 (2004) 119-126; Gautier, A. et al., Chem. Biol. 15 (2008) 128-136; and Protease-catalyzed formation of CN bond is used by Bordusa, F., Highlights in Bioorganic Chemistry (2004) 389-403).

[0449] Site-specific reactions and covalent couplings may also be achieved by selective reaction of the terminal amino acid with an appropriate modification reagent. The reactivity of N-terminal cysteines with benzonitrile may be used to achieve site-specific covalent bonds (see Ren, H. et al., Angew. Chem. Int. Ed. Engl. 48 (2009) 9658-9662). Native chemical ligation may also rely on a C-terminal cysteine ​​residue (see Taylor, E. Vogel; Imperiali, B, Nucleic Acids and Molecular Biology (2009), 22 (Protein Engineering), 65-96).

[0450] The moiety may also be a synthetic peptide or peptidomimetic. Polypeptides may be chemically synthesized and amino acids with orthogonal chemical reactivity may be integrated during such synthesis (see, for example, de Graaf, AJ et al., Bioconjug. Chem. 20 (2009) 1281-1295). To obtain a single labeled polypeptide, conjugates with 1:1 stoichiometry may be separated from other conjugation by-products by chromatography. This procedure may be facilitated by dye-labeled binding pair members and charged linkers. By using such types of labeled and highly negatively charged binding pair members, single binding polypeptides are easily separated from unlabeled polypeptides and polypeptides with one or more linkers, since the charge and molecular weight differences can be used for separation. Fluorescent dyes may be useful for purifying conjugates from unbound components, such as labeled monovalent binding agents.

[0451] V. Pharmaceutical Compositions The present invention provides compositions comprising a therapeutic agent (e.g., an LRRC4 family mimetic molecule, a FAM19A5 antagonist, or both; including nucleic acids, vectors, cells, or protein conjugates encoding and / or containing an LRRC4 family mimetic molecule and / or a FAM19A5 antagonist), excipients, or stabilizers in a physiologically acceptable carrier having a predetermined purity (Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, PA). Acceptable carriers, excipients, or stabilizers are nontoxic to the recipient in the amounts and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (e.g., octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl, or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol, and m-cresol); low molecular weight (less than about 10 residues) polypeptides; These include proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants such as TWEEN, PLURONICS, or polyethylene glycol (PEG).

[0452] In some aspects, pharmaceutical compositions useful in the present invention comprise any of the therapeutic agents described herein (e.g., an LRRC4 family mimetic molecule, a FAM19A5 antagonist, or both; including nucleic acids, vectors, cells, protein conjugates encoding and / or comprising an LRRC4 family mimetic molecule and / or a FAM19A5 antagonist) in a pharmaceutically acceptable carrier, and optionally one or more additional prophylactic or therapeutic agents. In some aspects, pharmaceutical compositions comprise any of the therapeutic agents described herein (e.g., an LRRC4 family mimetic molecule, a FAM19A5 antagonist, or both; including nucleic acids, vectors, cells, protein conjugates encoding and / or comprising an LRRC4 family mimetic molecule and / or a FAM19A5 antagonist) in a pharmaceutically acceptable carrier, and optionally one or more additional therapeutic agents. In some aspects, a therapeutic agent described herein is the only active ingredient contained in the pharmaceutical composition.

[0453] Pharmaceutically acceptable carriers used in parenteral formulations include aqueous vehicles, nonaqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, emulsifying agents, sequestering or chelating agents, and other pharma- ceutically acceptable substances. Examples of aqueous vehicles include sodium chloride injection, infusion injection, isotonic dextrose injection, sterile water injection, dextrose and lactate infusion injection. Nonaqueous parenteral vehicles include vegetable fixed oils, cottonseed oil, corn oil, sesame oil, and peanut oil. Antimicrobial agents in bacteriostatic or fungistatic concentrations may be added to parenteral formulations packaged in multi-dose containers including phenol or cresol, mercurial, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxybenzoic acid esters, thimerosal, benzalkonium chloride and benzethonium chloride. Isotonicity agents include sodium chloride and dextrose. Buffers include phosphates and citrates. Antioxidants include sodium bisulfate. Topical anesthetics include procaine hydrochloride. Suspending and dispersing agents include sodium carboxymethylcellulose, hydroxypropylmethylcellulose and polyvinylpyrrolidone. Emulsifying agents include polysorbate 80 (TWEEN 80). Sequestering or chelating agents for metal ions include EDTA. Pharmaceutical carriers also include ethyl alcohol, polyethylene glycol and propylene glycol for water-miscible vehicles; and sodium hydroxide, hydrochloric acid, citric acid or lactic acid for pH adjustment.

[0454] The pharmaceutical composition may be formulated for any route of administration to a subject. Specific examples of routes of administration include intranasal, oral, parenterally, intrathecally, intracerebroventricularly, pulmonary, subcutaneously, or intraventricularly. Parenteral administration, characterized by subcutaneous, intramuscular, or intravenous injection, is also contemplated herein. Injectables may be prepared in conventional forms, such as liquid solutions or suspensions, solid forms suitable for dissolving or suspending in liquid prior to injection, or emulsions. Injectables, solutions, and emulsions also contain one or more excipients. Suitable excipients are, for example, water, saline, dextrose, glycerol, or ethanol. In addition, if desired, the pharmaceutical composition to be administered may contain minor amounts of nontoxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents, stabilizers, solubility enhancers, and other formulating agents, such as, for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate, and cyclodextrins.

[0455] Formulations for parenteral administration of the therapeutic agents described herein (e.g., LRRC4 family mimetic molecules, FAM19A5 antagonists, or both; including nucleic acids, vectors, cells, protein conjugates encoding and / or comprising LRRC4 family mimetic molecules and / or FAM19A5 antagonists) include sterile solutions for injection, sterile dry soluble products such as lyophilized powders, sterile dry suspensions ready to be combined with a solvent immediately prior to use, including subcutaneous tablets, sterile suspensions for injection, sterile dry insoluble products ready to be combined with a vehicle immediately prior to use, and sterile emulsions. The solutions may be aqueous or non-aqueous.

[0456] For intravenous administration, suitable carriers include physiological saline or phosphate buffered saline (PBS), and solutions containing thickening and solubilizing agents such as glucose, polyethylene glycol, polypropylene glycol, and mixtures thereof.

[0457] Topical mixtures containing the therapeutic agent are prepared as described for local and systemic administration. The resulting mixtures may be solutions, suspensions, emulsions, etc., and may be formulated as creams, gels, ointments, emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes, foams, aerosols, irrigations, sprays, suppositories, bandages, skin patches, or other dosage forms suitable for topical administration.

[0458] Pharmaceutical compositions may be formulated as aerosols for topical application, such as inhalation (see, for example, U.S. Patent Nos. 4,044,126, 4,414,209 and 4,364,923). Such formulations for administration to the respiratory tract may be in the form of an aerosol or solution for nebulizers, or in the form of a fine powder for inhalation, alone or in combination with an inert carrier such as lactose. In this case, the particles of the formulation may have a diameter of less than about 50 microns, for example less than about 10 microns, in some aspects.

[0459] The pharmaceutical compositions may be formulated in the form of gels, creams and lotions for topical or local application, e.g., topical application to the skin and mucous membranes, application to the eye or for intrastriatal or intraspinal application. Topical administration is contemplated for transdermal delivery and / or administration to the eye or mucous membranes or for inhalation therapy. Nasal solutions of the antibody may be administered alone or together with other pharmaceutically acceptable excipients.

[0460] Transdermal patches, including iontophoretic and electrophoretic devices, are well known to those of skill in the art and may be used to administer the therapeutic agents described herein. For example, such patches are disclosed in U.S. Patent Nos. 6,267,983, 6,261,595, 6,256,533, 6,167,301, 6,024,975, 6,010715, 5,985,317, 5,983,134, 5,948,433, and 5,860,957.

[0461] In some aspects, the pharmaceutical compositions described herein are lyophilized powders that can be reconstituted for administration as solutions, emulsions, and other mixtures. They may also be reconstituted and formulated as solids or gels. Lyophilized powders are prepared by dissolving any of the therapeutic agents described herein or a pharma- ceutically acceptable derivative thereof in a suitable solvent. In some aspects, the lyophilized powders are sterile. The solvent can contain excipients that improve stability or other pharmacological components of the powder or reconstituted solution made from the powder. Excipients that can be used include, but are not limited to, dextrose, sorbitol, fructose, corn syrup, xylitol, glycerin, glucose, sucrose, or other suitable formulations. The solvent can also contain a buffer such as citrate, sodium or potassium phosphate, or other buffers known to those skilled in the art. In some aspects, the buffer is at about neutral pH. Subsequent sterile filtration of the solution following lyophilization under standard conditions known to those skilled in the art provides the desired formulation. In some aspects, the resulting solution may be divided into vials for lyophilization. Each vial may contain a single dose or multiple doses of any of the therapeutic agents described herein (e.g., LRRC4 family mimetic molecules, FAM19A5 antagonists, or both; including nucleic acids, vectors, cells, protein conjugates encoding and / or comprising LRRC4 family mimetic molecules and / or FAM19A5 antagonists). The lyophilized powder may be stored under appropriate conditions, such as at about 4° C. to room temperature.

[0462] Reconstitution of this lyophilized powder with water for injection provides a dosage form for parenteral administration. For reconstitution, the lyophilized powder is added to sterile water or other suitable carrier. The exact amount will vary depending on the compound selected. Such amounts may be empirically determined.

[0463] In some aspects, pharmaceutical compositions containing any of the therapeutic agents described herein may also be formulated to target specific tissues, receptors, or other areas of the body of a subject. For non-limiting examples of targeting methods, see, e.g., U.S. Patent Nos. 6,316,652, 6,274,552, 6,271,359, 6,253,872, 6,139,865, 6,131,570, 6,120,751, 6,071,495, 6,060,082, 6,048,736, 6,039,975, 6,004,534, 5,985,307, 5,972,366, 5,900,252, 5,840,674, 5,759,542, and 5,709,874.

[0464] Compositions to be used for in vivo administration can be sterilized, in some aspects, this can be accomplished, for example, by filtration through sterile filtration membranes.

[0465] VI. Nucleic Acids, Vectors, and Host Cells Additional aspects described herein relate to one or more nucleic acid molecules (also referred to herein as "nucleic acids" or derivatives thereof) encoding a therapeutic agent (e.g., a LRRC4 family mimetic molecule and / or a FAM19A5 antagonist described herein). The nucleic acids may be present in whole cells, a cell lysate, or in a partially purified or substantially pure form. In some aspects, the nucleic acid is a DNA sequence and / or an RNA sequence (e.g., mRNA). In some aspects, the nucleic acid comprises modified nucleotide analogs. A nucleic acid is "isolated" or "substantially pure" when it is purified from cellular components or other contaminants, e.g., other cellular nucleic acids (e.g., other chromosomal DNA, e.g., chromosomal DNA bound to naturally isolated DNA), by standard techniques well known in the art, including alkaline / SDS treatment, CsCl banding, column chromatography, restriction enzymes, agarose gel electrophoresis, and others (see F. Ausubel, et al., ed. (1987) Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York). In some aspects, the nucleic acid molecule may or may not contain intron sequences. In some aspects, the nucleic acid is a cDNA molecule. The nucleic acids described herein may be obtained using standard molecular biology techniques known in the art.

[0466] In some aspects, the present invention provides vectors comprising an isolated nucleic acid molecule encoding a therapeutic agent described herein (e.g., a LRRC4 family mimetic molecule and / or a FAM19A5 antagonist described herein). Suitable vectors of the present invention include, but are not limited to, expression vectors, viral vectors, and plasmid vectors. In some aspects, the vector is a viral vector.

[0467] As used herein, the term "expression vector" refers to any nucleic acid structure that contains the necessary elements for the transcription and translation of an inserted coding sequence, or, in the case of RNA viral vectors, the necessary elements for replication and translation, when introduced into an appropriate host cell. Expression vectors may include plasmids, phagemids, viruses, and derivatives thereof.

[0468] As used herein, "viral vector" includes, but is not limited to, nucleic acid sequences from the following viruses: retroviruses, e.g., Moloney murine leukemia virus, Harvey murine sarcoma virus, murine mammary tumor virus, and Rous sarcoma virus; lentiviruses; adenoviruses; adeno-associated viruses; SV40 viruses; polyomaviruses; Epstein-Barr viruses; papilloma viruses; herpes viruses; vaccinia viruses; polio viruses; and RNA viruses such as retroviruses. Certain viral vectors are based on non-cytopathic eukaryotic viruses in which non-essential genes have been replaced with the gene of interest. Non-cytopathic viruses include retroviruses, whose life cycle involves reverse transcription of genomic viral RNA into DNA followed by proviral integration into host cell DNA.

[0469] In some aspects, the vector is derived from an adeno-associated virus. In some aspects, the vector is derived from a lentivirus. Examples of lentivirus vectors are disclosed in WO9931251, WO9712622, WO9817815, WO9817816 and WO9818934, the entire contents of which are incorporated herein by reference.

[0470] Other vectors include plasmid vectors (see, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press, 1989). In recent years, plasmid vectors have proven particularly advantageous for transferring genes to cells in vivo because they are unable to replicate and integrate within the host genome. However, those plasmids that have promoters compatible with the host cell are capable of expressing peptides from genes operably encoded within the plasmid. Commonly used plasmids that are commercially available include pBR322, pUC18, pUC19, the various pcDNA plasmids, pRC / CMV, the various pCMV plasmids, pSV40, and pBlueScript. Additional examples of specific plasmids are pcDNA3.1, catalog number V79020; pcDNA3.1 / hygro, catalog number V87020; pcDNA4 / myc-His, catalog number V86320; and pBudCE4.1, catalog number V53220, all from Invitrogen, Carlsbad, Calif. Plasmids may also be custom designed using standard molecular biology techniques to remove and / or add specific segments of DNA.

[0471] Included in the invention are methods of producing the therapeutic agents disclosed herein (e.g., the LRRC4 family mimetic molecules and / or FAM19A5 antagonists described herein). In some aspects, such methods may include expressing the molecules (e.g., the LRRC4 family mimetic molecules and / or FAM19A5 antagonists described herein) in a cell that contains a nucleic acid molecule encoding the molecule. Host cells that contain these nucleotide sequences are included herein. Non-limiting examples of host cells that can be used include immortal hybridoma cells, NS / 0 myeloma cells, 293 cells, Chinese hamster ovary (CHO) cells, HeLa cells, human amniotic fluid-derived cells (CapT cells), COS cells, or combinations thereof.

[0472] VII. Kit The invention also provides kits comprising one or more therapeutic agents described herein (e.g., LRRC4 family mimetic molecules, FAM19A5 antagonists, or both; including nucleic acids, vectors, cells, protein conjugates encoding and / or comprising LRRC4 family mimetic molecules and / or FAM19A5 antagonists). As will be apparent from the present invention, in some aspects, kits comprising LRRC4 family mimetic molecules may be useful in treating diseases or disorders associated with increased FAM19A5 protein expression in a subject. In some aspects, provided herein is a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the pharmaceutical compositions described herein, optionally with instructions for use. In some aspects, the kit comprises a pharmaceutical composition described herein and any prophylactic or therapeutic agent as described herein.

[0473] The following examples are offered by way of illustration and not by way of limitation.

[0474] [Example] Example 1: Construction of LRRC4 family mimetic molecule-based ELISA assay To assess the ability of the LRRC4 mimetic molecules described herein to measure the amount of FAM19A5 protein in a sample, a sandwich ELISA assay was set up as shown in Figures 1A-1C. Briefly, 96-well plates were coated with one of the following: (i) a monoclonal anti-FAM19A5 antibody (Figure 1A); (ii) an Fc-linked LRRC4B peptide fragment (Figure 1B); and (iii) an LRRC4B peptide fragment not linked to Fc but instead linked to a His-tagged tobacco etch virus (TEV) protease sequence ("His-TEV-LRRC4B"). Biological samples (e.g., blood or cerebrospinal fluid) obtained from subjects were then added to wells of the plate and incubated as further described below. After incubation, HRP-linked monoclonal anti-FAM19A5 antibody was added to the relevant wells, and the amount of FAM19A5 protein was quantified using UV absorbance. Although this example uses an LRRC4B peptide fragment, it is clear from the invention that any of the LRRC4 mimetic molecules described herein (eg, an LRRC4 peptide fragment and / or an LRRC4C peptide fragment) may be used.

[0475] Example 2: Comparison of basal FAM19A5 protein levels in mice, rats, monkeys and humans Based on assays available in the art, blood FAM19A5 protein levels in both humans and mice (approximately 100 pg / mL and approximately 400 ng / mL, respectively) have been confirmed (see Lee et al., Diab Vasc Dis Res 16(6):530-538 (November 2019); and Wang et al., Circulation 138(1):48-63 (Jul. 2018), the entire contents of which are incorporated herein by reference). However, the accuracy of such assays is questionable, as Lee et al. (Mybiosource) and Wang et al. (R&D Systems) were unable to detect recombinant FAM19A5 protein produced in mammalian cells. As with many assays available in the art, such assays used anti-FAM19A5 antibodies generated using FAM19A5 protein purified from E. coli as an immunogen, indicating that such FAM19A5 protein can fold, unlike those not produced in mammalian cells. Therefore, to compare the methods described herein with methods available in the art, the His-TEV-LRRC4B-based ELISA assay described in Example 1 was used to quantify FAM19A5 protein levels in peripheral blood of naive mice (both wild-type and FAM19A5 knockout), rats, monkeys (both cynomolgus and marmosets) and humans. Human blood samples were purchased from BioIVT (Westbury, NY).

[0476] As shown in Figure 2, the blood FAM19A5 protein levels in naive mice (wild type), rats and monkeys were estimated to be approximately 100-200 pg / mL. In humans, the FAM19A5 protein level was confirmed to be approximately 150-400 pg / mL. No detectable amount of FAM19A5 protein was observed in FAM19A5 knockout animals to confirm the specificity of the analysis.

[0477] The above results demonstrate that, contrary to the published results mentioned above, the basal levels of FAM19A5 protein present in peripheral blood of various mammalian sources, including mice and humans, are generally similar.

[0478] Example 3: Analysis of FAM19A5 protein levels following administration of FAM19A5 antagonists To further evaluate the ability of the LRRC4 family mimetic molecules described herein to detect FAM19A5 protein present in samples, naive rats were administered a single intravenous dose of anti-FAM19A5 antibody (10 mg / kg or 50 mg / kg). Blood and cerebrospinal fluid (CSF) were then collected from the animals at various time points after antibody administration, and the levels of FAM19A5 protein present were evaluated using the His-TEV-LRRC4B-based ELISA assay described in Example 1.

[0479] As shown in FIG. 3, administration of anti-FAM19A5 antibody induced a rapid increase in blood FAM19A5 protein levels (within 6 hours of administration) in a dose- and time-dependent manner. The increase in blood FAM19A5 protein levels was somewhat delayed in animals treated with higher doses of anti-FAM19A5 antibody, but it should be noted that the FAM19A5 protein concentration observed immediately after antibody administration may not reflect the actual FAM19A5 concentration. As described herein, it was observed that high concentrations of administered anti-FAM19A5 antibody in the same presence may interfere with the accuracy of FAM19A5 ELISA. However, despite such a slight delay, rats administered higher doses generally had relatively higher amounts of FAM19A5 protein in the blood at most time points evaluated, compared to animals treated with low doses of anti-FAM19A5 antibody. A similar trend was observed from the CSF of the animals (see FIG. 4).

[0480] Given that 1) CSF anti-FAM19A5 antibody concentration partially reflects the concentration of antibody passing through the brain parenchyma, 2) the increase in anti-FAM19A5 antibody concentration precedes the increase in blood FAM19A5 protein levels, and 3) CSF anti-FAM19A5 antibody concentration and blood FAM19A5 protein concentration have a molar ratio of approximately 1:1 approximately 4 days after antibody administration, the above results imply that increased blood FAM19A5 protein levels are associated with anti-FAM19A5 antibody passing through the brain parenchyma.

[0481] Next, the His-TEV-LRRC4B-based ELISA assay described in Example 1 was used to evaluate blood FAM19A5 protein levels after weekly administration of anti-FAM19A5 antibody. Briefly, naive rats were intravenously administered anti-FAM19A5 antibody (10 mg / kg per dose) every week for 4 weeks. Blood was collected at various times during the antibody administration course to evaluate blood FAM19A5 protein levels. Blood anti-FAM19A5 antibody concentrations were also evaluated using an ELISA method using rabbit anti-human IgG Fc and HRP-conjugated goat anti-human IgG kappa light chain antibodies.

[0482] As shown in Figure 5, after each administration, the highest blood FAM19A5 protein level appeared about 3 days after administration and decreased until 7 days after administration. 2 hours after antibody administration, the FAM19A5 protein level was much lower than before administration, which means that the high antibody concentration present in the sample may again interfere with the FAM19A5 ELISA. Such results mean that the blood FAM19A5 protein level is a pharmacodynamic (PD) marker for the administered anti-FAM19A5 antibody.

[0483] For confirmation, cynomolgus monkeys were treated with a single intravenous dose of anti-FAM19A5 antibody (3 mg / kg, 10 mg / kg, or 30 mg / kg). Blood was collected at various time points after antibody administration, and the concentration of FAM19A5 protein was evaluated using the His-TEV-LRRC4B-based ELISA analysis described in Example 1. As shown in Figure 6, similar to what was observed from rat samples, higher doses of anti-FAM19A5 antibody induced higher blood FAM19A5 protein levels. Moreover, blood FAM19A5 protein levels were generally maintained in each treatment group until about 21 days after antibody administration.

[0484] The above results confirmed that blood FAM19A5 protein level is a suitable proximal PD marker for anti-FAM19A5 antibody action and provided evidence for target binding of the administered anti-FAM19A5 antibody.

[0485] Example 4: Analysis of FAM19A5 protein expression in traumatic brain injury-induced mice A mouse model of traumatic brain injury (TBI) was used to evaluate the diagnostic ability of the LRRC4 family mimetic molecules described herein. Briefly, 6-month-old female mice were deeply anesthetized with sodium pentobarbital (50 mg / kg). A pre-cooled iron bar was placed on the skull for 1 minute to perform cryogenic TBI. The skin of the mice was then sutured and they were kept in the same manner as normal mice (see Moon et al., Neuro Report 22:304-308 (2011)). Approximately 1 day after TBI induction, the animals were treated with anti-FAM19A5 antibody. As control groups, untreated TBI-induced mice and TBI-induced mice treated with human IgG antibody were used. Blood was collected from the animals at various time points, and the concentration of FAM19A5 protein in the peripheral blood was evaluated using the LRRC4 family mimetic molecule-based ELISA assay described in Example 1.

[0486] As shown in Figure 7, a significant increase in FAM19A5 protein expression was observed in the peripheral blood of control animals starting from about day 5 after TBI induction. Then, in animals treated with anti-FAM19A5 antibody, there was a rapid increase in peripheral blood FAM19A5 protein levels until about day 7 after TBI induction. Thereafter, FAM19A5 protein levels gradually decreased, and FAM19A5 protein levels returned to baseline (i.e., levels before anti-FAM19A5 antibody administration) 14 days after TBI induction.

[0487] The above results indicate that the LRRC4 family mimetic molecules described herein may be useful as diagnostic tools. The above results also indicate that anti-FAM19A5 antibody administration can accelerate FAM19A5 secretion from damaged brain to blood. Thus, it was confirmed that anti-FAM19A5 administration can induce higher secretion of FAM19A5 protein in patients with brain pathology than in normal individuals.

[0488] It should be understood that the Detailed Description section, and not the Abstract and Abstract sections, are intended to be used when analyzing the claims. The Abstract and Abstract sections may describe one or more but not all exemplary aspects of the invention contemplated by the inventor(s) and therefore are not intended to limit the scope of the invention and the appended claims in any respect.

[0489] The present invention has been described in detail with the aid of functional building blocks illustrating the implementation of certain functions and relationships thereof. The boundaries of such functional building blocks have been arbitrarily defined herein for the convenience of description. Alternative boundaries may be defined so long as the specified functions and relationships thereof are appropriately performed.

[0490] The foregoing description of specific aspects is intended to fully express the general characteristics of the inventive subject matter so that others may easily modify and / or adapt it to various adaptations of such specific aspects without undue experimentation, applying knowledge of the art. Thus, without departing from the general concept of the invention, such adaptations and modifications are intended to be within the meaning and range of equivalents of the presented aspects based on the description and guidance presented herein. The terms in this specification should be understood as being descriptive and not limiting, and therefore, the terms in this specification should be interpreted by skilled artisans in light of the description and guidance.

[0491] The breadth and scope of the present invention should not be limited by any of the above exemplary aspects, but should be defined only in accordance with the following claims and their equivalents.

[0492] All publications, patents, patent applications, internet sites and accession numbers / database sequences (including both polynucleotide and polypeptide sequences) cited herein are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, internet site or accession number / database sequence was specifically and individually indicated to be incorporated by reference. [Brief description of the drawings]

[0493] [Figure 1A] Figures 1A-1C provide schematic diagrams of different example approaches using a sandwich ELISA assay to measure FAM19A5 protein expression from a sample. Figure 1A shows the use of two different anti-FAM19A5 antibodies (Antibody A conjugated to HRP and Antibody B coated on an ELISA plate) in a sandwich ELISA assay to measure FAM19A5 protein expression from a sample. [Figure 1B]Figures 1A-1C provide schematic diagrams of different example approaches using a sandwich ELISA assay to measure FAM19A5 protein expression from a sample. Figure 1B shows the use of a LRRC4 family protein (LRRC4B) conjugated to Fc ("LRRC4B-hFc") and an anti-FAM19A5 antibody (Antibody A conjugated to HRP) in a sandwich ELISA assay to measure FAM19A5 protein expression from a sample. [Figure 1C] Figures 1A-1C provide schematic diagrams of different example approaches using a sandwich ELISA assay to measure FAM19A5 protein expression from a sample. Figure 1C shows the use of a LRRC4 family protein (e.g., LRRC4B) bound to a 6xHis tag, a Tobacco Etch Virus (TEV) protease sequence ("His-TEV-LRRC4B"), and an anti-FAM19A5 antibody (Antibody A conjugated to HRP) in a sandwich ELISA assay to measure FAM19A5 protein expression from a sample. [Diagram 2] FIG. 2 provides a comparison of FAM19A5 protein levels in the blood of mice (wild type and FAM19A knockout), rats, monkeys (cynomolgus and marmosets) and humans, as measured by the LRRC4 family mimetic molecule-based sandwich ELISA assay described in FIG. [Diagram 3] Figure 3 shows FAM19A5 protein levels in rats after treatment with anti-FAM19A5 antibody. Rats received a single intravenous dose of anti-FAM19A5 antibody at one of two doses, 10 mg / kg ("1") or 50 mg / kg ("2"). Blood was collected from the animals at various times after administration, and FAM19A5 protein levels were measured using the LRRC4 family mimetic molecule-based sandwich ELISA described in Figure 1. [Figure 4]Figure 4 shows FAM19A5 protein and anti-FAM19A5 antibody levels in blood and CSF, respectively, of rats after anti-FAM19A5 antibody treatment. Rats received a single intravenous dose of anti-FAM19A5 antibody (10 mg / kg). "Basal" = level before anti-FAM19A5 antibody administration. FAM19A5 protein levels were measured using the LRRC4 family mimetic molecule-based sandwich ELISA assay described in Figure 1. [Diagram 5] Figure 5 shows FAM19A5 protein and anti-FAM19A5 antibody levels in blood of rats administered anti-FAM19A5 antibody intravenously every week. Rats received a total of four doses (10 mg / kg per dose). Anti-FAM19A5 antibody levels were measured using an ELISA method using rabbit anti-human IgG Fc and HRP-conjugated goat anti-human IgG kappa light chain antibodies. FAM19A5 protein levels were measured using the LRRC4 family mimetic molecule-based sandwich ELISA assay described in Figure 1. [Figure 6] Figure 6 shows FAM19A5 protein levels in monkey blood at various time points after anti-FAM19A5 administration. Anti-FAM19A5 antibodies were administered at three different doses (3 mg / kg ("1"), 10 mg / kg ("2") or 30 mg / kg ("3")) and monkeys were administered a single antibody intravenously. FAM19A5 protein levels were measured using the LRRC4 family mimetic molecule-based sandwich ELISA assay described in Figure 1. [Figure 7] Figure 7 shows FAM19A5 protein levels in blood of traumatic brain injury (TBI)-induced mice treated with anti-FAM19A5 antibody. Mice were administered anti-FAM19A5 ("2") antibody twice (see arrows) (30 mg / kg per dose). Untreated TBI-induced mice and TBI-induced mice administered human IgG antibody ("1") served as control groups. FAM19A5 protein levels were measured using the LRRC4 family mimetic molecule-based sandwich ELISA assay described in Figure 1. Data are shown as mean ± SEM.

Claims

1. (i) contacting a biological sample from a subject with a leucine-rich containing 4 ("LRRC4") family mimetic molecule capable of specifically binding to family with sequence similarity 19, member A5 ("FAM19A5") protein; and (ii) measuring the expression level of FAM19A5 protein in the biological sample ("FAM19A5 protein expression level"); The measuring step comprises quantifying the amount of an LRRC4 family mimetic molecule-FAM19A5 complex, and the method for determining the expression level of FAM19A5 protein in a subject in need thereof.

2. The method of claim 1, further comprising confirming or predicting whether the subject is suffering from a disease or disorder.

3. 3. The method of claim 2, wherein the disease or disorder is associated with increased FAM19A5 protein expression levels.

4. The subject is a subject that is compatible with or responsive to treatment with an antagonist against the FAM19A5 protein (a "FAM19A5 antagonist"); (a) the subject is suitable for treatment if the FAM19A5 protein expression level is increased compared to the corresponding expression level in a subject not suffering from the disease or disorder; or (b) the subject is responsive to treatment if the FAM19A5 protein expression level is decreased compared to the corresponding expression level in the subject before treatment.

5. (a) The FAM19A5 protein expression level is measured by any of enzyme-linked immunosorbent assay (ELISA), immunohistochemistry, Western blotting, radioimmunoassay, radical-immunodiffusion, immunoprecipitation assay, Ouchterlony immunodiffusion method, rocket immunoelectrophoresis, and tissue immunostaining. immunostaining method), complement fixation assay, FACS, protein chip, SIMOA technology or a combination thereof; (b) the biological sample comprises blood, cerebrospinal fluid (CSF), serum, plasma, tissue, cell culture medium, saliva, urine, or a combination thereof; or (c) The method of claim 1, wherein both (a) and (b) are satisfied.

6. 10. The method of claim 1, wherein the contacting and measuring steps are performed outside the body.

7. The method of claim 4, wherein the therapeutic agent comprises an antagonist to FAM19A5 ("FAM19A5 antagonist"), a leucine-rich containing 4 ("LRRC4") family mimetic molecule, or both.

8. The method of claim 7, wherein the FAM19A5 antagonist comprises an antibody or its antigen-binding portion, an antisense oligonucleotide, an siRNA, an shRNA, an miRNA, a dsRNA targeting FAM19A5, an aptamer, a PNA, a vector containing the same, or a combination thereof.

9. 2. The method of claim 1, wherein the LRRC4 family mimetic molecule is (a) not an antibody or an antigen-binding portion thereof, (b) a polypeptide, or (c) a small molecule.

10. The method of claim 9, wherein the LRRC4 protein family mimetic molecule is a polypeptide comprising a domain of an LRRC4 protein family member, wherein the domain ("FAM19A5 binding domain") is capable of binding to FAM19A5 protein, and wherein the polypeptide is shorter than the corresponding full-length LRRC4 protein family member (SEQ ID NO: 4; SEQ ID NO: 5; or SEQ ID NO: 6).

11. The method of claim 9, wherein the LRRC4 family mimetic molecule is a polypeptide comprising an amino acid sequence (from N-terminus to C-terminus) having the following chemical formula: (a) A-(T / S)-B (Chemical Formula I): wherein (i) A comprises X1-(T / S)-(Y / F)-F-X5; X1 is tyrosine (Y), phenylalanine (F), valine (V), leucine (L), or isoleucine (I); (T / S) is threonine (T) or serine (S); (Y / F) is tyrosine (Y) or phenylalanine (F); X5 is any amino acid; (ii) B comprises (V / I)-TV-(E / V); (V / I) is valine (V) or isoleucine (I); (E / V) is glutamic acid (E) or valine (V); (b) A-(T / S)-B (chemical formula I): wherein (i) A comprises (Y / W / M)-(T / Y)-(Y / W)-(F / Y / W)-(T / Y); (Y / W / M) is tyrosine (Y), tryptophan (W), or methionine (M); (T / Y) is threonine (T) or tyrosine (Y); (Y / W) is tyrosine (Y) or tryptophan (W); (F / Y / W) is phenylalanine (F), tyrosine (Y), or tryptophan (W); (ii) B comprises X7-(T / S / Y)-X9-X10; X7 is valine (V), tyrosine (Y), phenylalanine (F), leucine (L), tryptophan (W), or methionine (M); (T / S / Y) is threonine (T), serine (S), or tyrosine (Y); X9 is valine (V), isoleucine (I), tyrosine (Y), phenylalanine (F), leucine (L), tryptophan (W), or methionine (M); X10 is glutamic acid (E), aspartic acid (D), isoleucine (I), tyrosine (Y), phenylalanine (F), methionine (M), or tryptophan (W); (c) X1-X2-X3-F-X5-T-X7-TV-X10 (Formula II): wherein X1 is Y, F, V, L, or I; X2 is T or S; X3 is Y or F; X5 is any amino acid; X7 is V or I; and / or X10 is E or V; or (d) X1-X2-X3-X4-X5-X6-X7-X8-X9-X10 (Formula VI): wherein X1 is Y, F, V, L, I, W, or M; X2 is T, S, or Y; X3 is Y, F, or W; X4 is F, Y, or W; X5 is any amino acid, for example, T, S, or Y; X6 is T, S, or Y; X7 is V, I, Y, F, L, W, or M; X8 is T, S, or Y; X9 is V, I, Y, F, L, W, or M; and / or X10 is E, D, V, I, Y, F, M, or W.

12. The method of claim 11, characterized in that the LRRC4 family mimetic molecule is a polypeptide comprising: (a) the amino acid sequence represented by SEQ ID NO: 29 (YTYFTTVTVE), (b) the amino acid sequence represented by SEQ ID NO: 20 (NYSFFTTVTVETTEISPEDTTRK), (c) the amino acid sequence represented by SEQ ID NO: 21 (NFSYFSTVTVETMEPSQDERTTR), or (d) the amino acid sequence represented by SEQ ID NO: 18 (GYTYFTTVTVETLETQPGEE).

13. The LRRC4 family mimetic molecule is (a) an amino acid sequence having at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% sequence identity with the amino acid sequence set forth in SEQ ID NO: 29 (YTYFTTVTVE); or (b) a polypeptide comprising an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or 99% identical to the amino acid sequence represented by SEQ ID NO:5, SEQ ID NO:4, or SEQ ID NO:6, and that contains at least one amino acid modification relative to the amino acid sequence represented by SEQ ID NO:5, SEQ ID NO:4, or SEQ ID NO:6, respectively.

14. The LRRC4 family mimetic molecule is (a) does not contain the transmembrane or intracellular domain of an LRRC4 protein family member; (b) further comprising one or more additional amino acids at the N-terminus, C-terminus, or both the N-terminus and C-terminus of the LRRC4 family mimetic molecule; (c) the N-terminus, the C-terminus, or both the N-terminus and the C-terminus contain modifications that increase the stability of the LRRC4 family mimetic molecule; or (d) The method of claim 1, characterized in that it is any combination of (a) to (c).

15. The method of claim 1, wherein the LRRC4 family mimetic molecule further comprises a half-life extending moiety.

16. The LRRC4 family mimetic molecule is (a) A small molecule of formula (I): 【Chemistry 1】 or a pharmaceutically acceptable salt thereof; where (i) R １ , R ２ and R ３ is hydrogen (hydrogen), fluoro (fluoro), chloro (chloro), bromo (bromo), methyl (methyl), ethyl (ethyl), n-propyl (n-propyl), iso-propyl (iso-propyl), n-butyl (n-butyl), iso-butyl (iso-butyl), t-butyl (t-butyl), n-pentyl (n-pentyl), iso-pentyl (iso-pentyl), fluoromethyl (fluo fluoromethyl), difluoromethyl, trifluoromethyl, 2-fluoroethyl, 1-fluoroethyl, 2,2-difluoroethyl, 1,2-difluoroethyl, 1,1-difluoroethyl, 2,2,2-trifluoroethyl, methoxy, ethoxy, n-propyloxy, isopropyloxy, n-butoxy, trifluoromethoxy, difluoromethoxy, fluoromethoxy , acetyl, propionyl, n-butanoyl, iso-butanoyl, n-pentanoyl, nitro, amino, N-methylamino, N-ethylamino, N-n-propylamino, N,N-dimethylamino,N-dimethylamino), N-acetylamino, N-propionylamino, N-(trifluoroacetyl)amino, formyl, hydroxy, methylthio, ethylthio, n-propylthio independently selected from propylthio, methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, phenyl, hydroxymethyl, 1-hydroxyethyl, and 2-hydroxyethyl; (ii) 【Chemistry 2】 is a single or double bond; (iii) Z is a linear or branched (C １ -C ８ ) alkyl, linear or branched (C ２ -C ８ ) alkenyl, linear or branched (C ２ -C ８ ) alkynyl, (C ３ -C ８ ) cycloalkyl, (C ５ -C ８ ) cycloalkenyl, (3-8 membered) heterocycloalkyl, (C ７ -C １４ ) bicycloalkyl, (C ７ -C １４ ) bicycloalkenyl, (7-14 membered) heterobicycloalkyl, (C ６ -C １０ )aryl, (5-10 membered)heteroaryl, and —CH—C(O)—CH═CH—Q, where Q is (C ３ -C ８ ) cycloalkyl, (C ５ -C ８ ) cycloalkenyl, (3-8 membered) heterocycloalkyl, (C ６ -C １０ )aryl, and (5-6 membered)heteroaryl; each cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl is (C １ -C ６ ) alkoxy, (C １ -C ６ ) alkyl, halo, (C １ -C ６ ) optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from haloalkoxy, nitro, amino, N-methylamino, N-ethylamino, N—N-propylamino, N,N-dimethylamino, formyl, and hydroxy; (iv) L is a single, double, or triple bond; (b) A small molecule of formula (II): 【Transformation 3】 or a pharmaceutically acceptable salt thereof; wherein (i) R 1 , R 2 and R 3 are hydrogen, fluoro, chloro, bromo, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, t-butyl, n-pentyl, iso-pentyl, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 1-fluoroethyl, 2,2-difluoroethyl, 1,2-difluoroethyl, 1,1-difluoroethyl, 2,2,2-trifluoroethyl, methoxy, ethoxy, n-furopyroxy, iso-furopyroxy, n-butoxy, trifluoromethoxy, difluoromethoxy, fluoromethyl; independently selected from methyl, acetyl, propionyl, n-butanoyl, iso-butanoyl, n-pentanoyl, nitro, amino, N-methylamino, N-ethylamino, N-n-propylamino, N,N-dimethylamino, N-acetylamino, N-propionylamino, N-(trifluoroacetyl)amino, formyl, hydroxy, methylthio, ethylthio, n-propylthio, methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, phenyl, hydroxymethyl, 1-hydroxyethyl, and 2-hydroxyethyl; (ii) Z is selected from linear or branched (C 1 -C 8 ) alkyl, linear or branched (C 2 -C 8 ) alkenyl, linear or branched (C 2 -C 8 ) alkynyl, (C 3 -C 8 ) cycloalkyl, (C 5 -C 8 ) cycloalkenyl, (3-8 membered) heterocycloalkyl, (C 7 -C 14 ) bicycloalkyl, (C 7 -C 14 ) bicycloalkenyl, (7-14 membered) heterobicycloalkyl, (C 6 -C 10 ) aryl, (5-10 membered) heteroaryl, and —CH═CH-Q; and Q is selected from (C 3 -C 8 ) cycloalkyl, (C 5 -C 8 ) cycloalkenyl, (3-8 membered) heterocycloalkyl, (C 6 -C 10 )aryl, and (5-6 membered)heteroaryl; each cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl is optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from (C 1 -C 6 )alkoxy, (C 1 -C 6 )alkyl, halo, (C 1 -C 6 )haloalkoxy, nitro, amino, N-methylamino, N-ethylamino, N—N-propylamino, N,N-dimethylamino, formyl, and hydroxy; (iii) L is a single, double, or triple bond; or (c) A small molecule of formula (III): 【Chemistry 4】 or a pharmaceutically acceptable salt thereof; wherein (i) R 1 , R 2 and R 3 are hydrogen, fluoro, chloro, bromo, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, t-butyl, n-pentyl, iso-pentyl, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 1-fluoroethyl, 2,2-difluoroethyl, 1,2-difluoroethyl, 1,1-difluoroethyl, 2,2,2-trifluoroethyl, methoxy, ethoxy, n-furopyroxy, iso-furopyroxy, n-butoxy, trifluoromethoxy, difluoromethoxy, fluoromethyl; independently selected from methyl, acetyl, propionyl, n-butanoyl, iso-butanoyl, n-pentanoyl, nitro, amino, N-methylamino, N-ethylamino, N-n-propylamino, N,N-dimethylamino, N-acetylamino, N-propionylamino, N-(trifluoroacetyl)amino, formyl, hydroxy, methylthio, ethylthio, n-propylthio, methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, phenyl, hydroxymethyl, 1-hydroxyethyl, and 2-hydroxyethyl; (ii) Z is selected from linear or branched (C 1 -C 8 ) alkyl, linear or branched (C 2 -C 8 ) alkenyl, linear or branched (C 2 -C 8 ) alkynyl, —Y—(C 3 -C 8 ) cycloalkyl, —Y—(C 5 -C 8 ) cycloalkenyl, —Y-(3-8 membered) heterocycloalkyl, —Y—(C 7 -C 14 ) bicycloalkyl, —Y—(C 7 -C 14 ) bicycloalkenyl, —Y-(7-14 membered) heterobicycloalkyl, —Y—(C 6 -C 10 ) aryl, and —Y-(5-10 membered) heteroaryl; and Y is a bond or C 1 -C 3 straight chain or branched alkylene, wherein said cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from (C 1 -C 6 )alkoxy, (C 1 -C 6 )alkyl, halo, (C 1 -C 6 )haloalkoxy, nitro, amino, N-methylamino, N-ethylamino, N—N-propylamino, N,N-dimethylamino, formyl, and hydroxy; (iii) L is a single, double, or triple bond; (iv) n is 0 or 1.

17. The LRRC4 family mimetic molecule is a mimetic molecule selected from the following structural formulas: (a) 【Transformation 5】 or a pharmaceutically acceptable salt thereof; (b) 【Transformation 6】 or a pharmaceutically acceptable salt thereof; (c) 【Transformation 7】 or a pharmaceutically acceptable salt thereof.

18. 3. The method of claim 2, wherein the disease or disorder comprises amyotrophic lateral sclerosis (ALS), Alzheimer's disease, glaucoma, diabetic retinopathy, neuropathic pain, spinal cord injury, traumatic brain injury, stroke, Parkinson's disease, or a combination thereof.

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