Use of anti-FAM19A5 antibodies

Abstract

The present disclosure relates to the pharmaceutical use of antagonists (e.g., antibodies or antigen-binding portions thereof) that specifically bind to FAM19A5 for the treatment of retinopathy (e.g., diabetic retinopathy) and / or maculopathy (e.g., age-related macular degeneration) in an individual in need thereof.

Description

[0001] This application is a divisional application of a patent application with application number 201980075558.7, application date October 15, 2019, and invention name “Use of anti-FAM19A5 antibodies”.

[0002] Cross-references to related patent applications

[0003] This PCT application claims priority to U.S. Provisional Patent Application No. 62 / 746,194 filed on October 16, 2018, which is incorporated herein by reference in its entirety and becomes a part of the present invention.

[0004] See electronic sequence listing

[0005] An electronic version of the sequence listing in ASCII text file (name: 3763.012PC01_SeqListing_ST25.txt; size: 261609 bytes; creation date: October 13, 2019) is submitted with this application, and its contents are incorporated herein by reference as a whole and become a part of the present invention. Technical Field

[0006] The present disclosure provides methods of treating retinopathy (e.g., diabetic retinopathy) and / or macular degeneration (e.g., age-related macular degeneration) in an individual (e.g., a human) using antagonists to sequence similar family 19, member A5 (FAM19A5) (e.g., anti-FAM19A5 antibodies) or compositions comprising such antagonists. Background Art

[0007] Retinopathy such as diabetic retinopathy (DR), also known as diabetic eye disease, is caused by damage to the small blood vessels and neurons of the retina. DR is the most common cause of vision loss in the working-age population (e.g., 20-74 years old) worldwide. Lee R. et al., Eye Vis (Lond) 2:17 (2015). In recent years, the incidence of DR has increased dramatically with the increase in life expectancy of diabetic patients. Roughly speaking, about 50% of diabetic patients develop DR 10 years after the onset of diabetes, 70% of diabetic patients develop DR 20 years after the onset of diabetes, and 90% of diabetic patients develop DR 30 years after the onset of diabetes. It is estimated that by 2030, the number of diabetic patients in the 20-79 age group worldwide will almost double, which means that the number of people diagnosed with DR will increase further. Nentwich MM and Ulbig MW, World J Diabetes 6(3):489-499 (2015).

[0008] Although there are treatment options available that can help manage symptoms associated with DR, these treatments are not curative and can have adverse side effects. For example, laser therapy can cause severe discomfort, permanent retinal scarring, and decreased peripheral, color, and night vision during treatment. See worldwideweb.aao.org / munnerlyn-laser-surgery-center / laser-treatment-of-proliferative-nonproliferative-. Therefore, there is a need for alternative treatment options for diabetic retinopathy.

[0009] Age-related macular degeneration (AMD) is another retinal disease that affects the eye, specifically the central part of the eye (the macula). AMD is caused by hardening of the arteries that nourish the retina, which deprives the retinal tissue of oxygen and leads to decreased central vision. Gheorghe A. et al., Rom J Ophthalmol 59(2):74-77 (2015). AMD is the most common form of macular degeneration, affecting approximately 11 million people in the United States alone and 170 million people worldwide. AMD is the leading cause of visual disability in industrialized countries and the third leading cause of visual disability worldwide. Pennington KL and DeAngelis MM, Eye Vis (Lond) 3:34 (2016). Aging is the greatest risk factor; therefore, the number of people with AMD in the United States is expected to increase to 22 million by 2050 and to 288 million worldwide by 2040. Wong WL et al., Lancet Glob Health 2: e106-e116 (2014).

[0010] If diagnosed early, AMD can be treated with a variety of treatment options, including vitamin supplements (e.g., antioxidants), dietary changes (e.g., a low-fat diet), anti-angiogenic drugs (e.g., anti-vascular endothelial growth factor (VEGF) antibodies), laser therapy, surgery (e.g., subretinal surgery), and retinal transposition. Holz FG et al., J Clin Invest 124(4):1430-1438 (2014). However, there is currently no known cure for AMD, and many treatment options may not be suitable for all patients with AMD. For example, many patients with choroidal neovascularization (CNV) (i.e., wet AMD) are not suitable for laser therapy because the CNV is too large for laser treatment or its location cannot be accurately determined. In addition, even with treatment, many patients with AMD will gradually lose central vision. Virgili G. and Bini A., Cochrane Database Syst, Rev. 3:CD004763 (2007). Therefore, there is a need for more effective and comprehensive AMD treatment options. Summary of the invention

[0011] The present invention provides a method for improving the retinal potential in the retina of an individual in need thereof compared to a reference value (e.g., the corresponding value of the individual before the administration of a FAM19A5 antagonist, or the corresponding value of the individual without the administration of a FAM19A5 antagonist), comprising administering an antagonist to a protein of family 19, member A5 ("FAM19A5") of sequence similarity to the individual ("FAM19A5 antagonist"). In certain aspects, the improvement of the retinal potential comprises an increase in A waves, B waves, oscillatory potentials, or any combination thereof.

[0012] In some aspects, the individual suffers from a disease or condition associated with decreased retinal potential. In some aspects, the disease or condition refers to diabetic retinopathy. In other aspects, the disease or condition refers to age-related macular degeneration of the retina.

[0013] The present invention also provides a method of treating retinopathy and / or maculopathy in a patient in need thereof, comprising administering a subject FAM19A5 antagonist.

[0014] In some aspects, retinopathy refers to diabetic retinopathy. In some aspects, diabetic retinopathy refers to non-proliferative diabetic retinopathy (NPDR). In other aspects, diabetic retinopathy refers to proliferative diabetic retinopathy. In some aspects, diabetic retinopathy is associated with decreased retinal potential within the individual retina, pericyte loss, formation of acellular capillaries, vascular congestion, or any combination thereof.

[0015] In certain aspects, maculopathy refers to diabetic macular edema. In certain aspects, diabetic macular edema is associated with decreased retinal potential, pericyte loss, formation of acellular capillaries, vascular congestion, or any combination thereof within the retina of the individual.

[0016] In some aspects, compared with a reference value (e.g., the corresponding value of the individual before the FAM19A5 antagonist is applied, or the corresponding value of the individual when the FAM19A5 antagonist is not applied), the FAM19A5 antagonist can improve the retinal potential of the individual. In some aspects, the improvement of the retinal potential includes an increase in A waves, B waves, oscillatory potentials, or any combination thereof. In some aspects, the FAM19A5 antagonist reduces the loss of pericytes in the individual retina. In some aspects, the FAM19A5 antagonist is used to reduce and / or inhibit the formation of acellular capillaries in the individual retina. In other aspects, the FAM19A5 antagonist is used to reduce and / or inhibit vascular congestion in the individual retinal nerve fiber layer.

[0017] The present disclosure also provides a method for treating macular degeneration in an individual in need thereof, comprising administering an antagonist to the FAM19A5 protein to the individual.

[0018] In some aspects, macular degeneration refers to age-related macular degeneration. In some aspects, age-related macular degeneration refers to dry (atrophic) macular degeneration. In other aspects, age-related macular degeneration refers to wet (neovascular or exudative) macular degeneration. In some aspects, age-related macular degeneration refers to (i) early AMD, (ii) mid-term AMD or (iii) late AMD (geographic atrophy). In more aspects, age-related macular degeneration is related to accumulation or aggregation of drusen in the individual retina, neovascularization, retinal potential reduction, astrocyte formation or any combination thereof.

[0019] In some aspects, CD31 and / or VEGF expression reduction indicates that FAM19A5 antagonists are used to reduce and / or inhibit neovascularization in the individual retina. In some aspects, CD31 and / or VEGF expression reduction indicates that FAM19A5 antagonists can reduce and / or inhibit neovascularization in the retina of mice with laser-induced choroidal neovascularization. In more aspects, wherein, compared with a reference value (e.g., the corresponding value of the individual before the application of the FAM19A5 antagonist, or the corresponding value of the individual without the application of the FAM19A5 antagonist), the FAM19A5 antagonist reduces the platelet-derived growth factor (PDGF) expression in the nerve fiber layer and ganglion cell layer of the individual retina. In other aspects, compared with a reference value (e.g., the corresponding value of the individual before the application of the FAM19A5 antagonist, or the corresponding value of the individual without the application of the FAM19A5 antagonist), the FAM19A5 antagonist can improve the retinal potential of the individual. In some aspects, the improvement of the retinal potential includes an increase in B waves. In certain aspects, a FAM19A5 antagonist can reduce and / or inhibit the accumulation or aggregation of drusen in the retina of an individual.

[0020] In certain aspects, the FAM19A5 antagonist is an antisense oligonucleotide, siRNA, shRNA, miRNA, dsRNA targeting FAM19A5, aptamer, PNA, or a vector comprising the same.

[0021] In some aspects, the FAM19A5 antagonist is an antibody or antigen-binding portion thereof that specifically binds to the FAM19A5 protein ("anti-FAM19A5 antibody"). In some aspects, the anti-FAM19A5 antibody exhibits properties selected from the following: (a) binding to soluble human FAM19A5 as measured by enzyme-linked immunosorbent assay (ELISA), K D Less than or equal to 10 nM; (b) Binding to membrane-bound human FAM19A5 as measured by ELISA, K D Less than or equal to 10 nM; (c) Both (a) and (b).

[0022] In certain aspects, the anti-FAM19A5 antibody cross-competes with a reference antibody for binding to a human FAM19A5 epitope, wherein the reference antibody is selected from the group consisting of the antibodies in Tables 2-5. In certain aspects, the reference antibody comprises a heavy chain CDR1, CDR2 and CDR3 and a light chain CDR1, CDR2 and CDR3, (i) wherein the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 14, the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 15, the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 16, the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 26, the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 27, and the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 28; or (ii) wherein the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 17, the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 18, the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 19, the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 29, the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 30, and the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: The amino acid sequence shown in NO:31.

[0023] In certain aspects, the anti-FAM19A5 antibody specifically binds the same epitope of FAM19A5 as a reference antibody, wherein the reference antibody is selected from the group consisting of Tables 2-5. In certain aspects, the reference antibody comprises a heavy chain CDR1, CDR2 and CDR3 and a light chain CDR1, CDR2 and CDR3, (i) wherein the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 14, the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 15, the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 16, the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 26, the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 27, and the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 28; or (ii) wherein the heavy chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 17, the heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 18, the heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 19, the light chain CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 29, the light chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 30, and the light chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 31. The amino acid sequence shown in NO:31.

[0024] In certain aspects, the anti-FAM19A5 antibody specifically binds at least one FAM19A5 epitope, ie, SEQ ID NO:6 or SEQ ID NO:9.

[0025] In certain aspects, the anti-FAM19A5 antibody comprises heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3, (i) wherein the heavy chain CDR1 comprises CDR1, CDR1 is selected from the group consisting of CDR1 in Table 2; (ii) wherein the heavy chain CDR2 comprises CDR2, CDR2 is selected from the group consisting of CDR2 in Table 2; (iii) wherein the heavy chain CDR3 comprises CDR3, CDR3 is selected from the group consisting of CDR3 in Table 2; (iv) wherein the light chain CDR1 comprises CDR1, CDR1 is selected from the group consisting of CDR1 in Table 3; (v) wherein the light chain CDR2 comprises CDR2, CDR2 is selected from the group consisting of CDR2 in Table 3; and / or (vi) wherein the light chain CDR3 comprises CDR3, CDR3 is selected from the group consisting of CDR3 in Table 3.

[0026] In certain aspects, the anti-FAM19A5 antibody comprises heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3, (i) wherein the heavy chain CDR1 comprises the amino acid sequence shown in SEQ ID NO: 14 or SEQ ID NO: 17; (ii) wherein the heavy chain CDR2 comprises the amino acid sequence shown in SEQ ID NO: 15 or SEQ ID NO: 18; (iii) wherein the heavy chain CDR3 comprises the amino acid sequence shown in SEQ ID NO: 16 or SEQ ID NO: 19; (iv) wherein the light chain CDR1 comprises the amino acid sequence shown in SEQ ID NO: 26 or SEQ ID NO: 29; (v) wherein the light chain CDR2 comprises the amino acid sequence shown in SEQ ID NO: 27 or SEQ ID NO: 30; and / or (vi) wherein the light chain CDR3 comprises the amino acid sequence shown in SEQ ID NO: 28 or SEQ ID NO: 31.

[0027] In certain aspects, the anti-FAM19A5 antibody comprises heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3, wherein the heavy chain CDR1, CDR2 and CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 14, 15 and 16, respectively, and the light chain CDR1, CDR2 and CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 26, 27 and 28, respectively. In other aspects, the anti-FAM19A5 antibody comprises heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3, wherein the heavy chain CDR1, CDR2 and CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 17, 18 and 19, respectively, and the light chain CDR1, CDR2 and CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 29, 30 and 31, respectively.

[0028] In certain aspects, the anti-FAM19A5 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), (i) wherein the VH comprises the amino acid sequence shown in SEQ ID NO: 36 or SEQ ID NO: 37; or (ii) wherein the VL comprises the amino acid sequence shown in SEQ ID NO: 40 or SEQ ID NO: 41. In certain aspects, the VH comprises the amino acid sequence of SEQ ID NO: 36, and the VL comprises the amino acid sequence shown in SEQ ID NO: 40. In other aspects, the VH comprises the amino acid sequence of SEQ ID NO: 37, and the VL comprises the amino acid sequence shown in SEQ ID NO: 41.

[0029] In certain aspects, the anti-FAM19A5 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises an amino acid sequence that is 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%, at least about 99% or about 100% identical to the amino acid sequence set forth as SEQ ID NO: 36 or SEQ ID NO: 37. In certain aspects, the anti-FAM19A5 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VL comprises an amino acid sequence that is 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%, at least about 99% or about 100% identical to the amino acid sequence set forth as SEQ ID NO: 40 or SEQ ID NO: 41.

[0030] In certain aspects, the anti-FAM19A5 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), (i) wherein the VH comprises an amino acid sequence that is 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%, at least about 99% or about 100% identical to the amino acid sequence set forth as SEQ ID NO: 36 or SEQ ID NO: 37; and (ii) wherein the VL comprises an amino acid sequence that is 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%, at least about 99% or about 100% identical to the amino acid sequence set forth as SEQ ID NO: 40 or SEQ ID NO: 41.

[0031] In some aspects, the anti-FAM19A5 antibody comprises Fab, Fab', F(ab')2, Fv or single-chain Fv (scFv). In some aspects, the anti-FAM19A5 antibody is selected from the group consisting of IgG1, IgG2, IgG3, IgG4 and variants thereof. In some aspects, the anti-FAM19A5 antibody is IgG2, IgG4 or a combination thereof. In more aspects, the anti-FAM19A5 antibody comprises an IgG2 / IgG4 isotype antibody. In some aspects, the anti-FAM19A5 antibody comprises a constant region without Fc function. In some aspects, the anti-FAM19A5 antibody is a chimeric antibody, a human antibody or a humanized antibody.

[0032] In certain aspects, the FAM19A5 antagonist is linked to a molecule having a second binding moiety, thereby forming a bispecific molecule. In certain aspects, the FAM19A5 antagonist is linked to an agent, thereby forming an immunoconjugate.

[0033] In some aspects, the FAM19A5 antagonist disclosed in the present invention is formulated with a pharmaceutically acceptable carrier. In some aspects, the FAM19A5 antagonist is administered by intraocular administration. In some aspects, intraocular administration includes intravitreal administration. In some aspects, the FAM19A5 antagonist is administered by intravenous injection.

[0034] In certain aspects, the methods disclosed herein further comprise administering a therapeutic agent. In certain aspects, the individual receiving the disclosed treatment is a human.

[0035] The present disclosure also provides a pharmaceutical composition for preventing and treating retinopathy and / or maculopathy, comprising a pharmaceutically effective amount of an antagonist against the FAM19A5 protein. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1A , 1B Figures 1 and 1C show the effect of anti-FAM19A5 antibody on retinal potential recovery in 25-week-old db / db mice. Mice were treated with human IgG control antibody ("IgG") or anti-FAM19A5 antibody (clone "1-65" or "3-2"). Normal (i.e., non-diabetic) mice were used as controls. Each group of mice was exposed to three different flash intensities (-1, 0.2, and 0.8 log cds / m 2 ) under white light, and then measure the A wave (μV) ( Figure 1A )、B wave (μV) ( Figure 1B ) and oscillating potential (μV) ( Figure 1C ). Data are expressed as mean ± SD.

[0037] Figure 2A and 2B The protective effect of anti-FAM19A5 antibody on retinal microvessels in 25-week-old db / db mice is shown. Mice were treated with human IgG control antibody ("IgG") or anti-FAM19A5 antibody (clone "1-65" or "3-2"). Normal (ie, non-diabetic) healthy mice were used as controls. Figure 2A Shown are microscopic images (100× magnification) of representative retinal areas of mice in various treatment groups after periodic acid-Schiff (PAS) staining: (i) normal (i.e., non-diabetic) (upper left panel), (ii) human IgG control (upper right panel), and (iii) anti-FAM19A5 antibody (clone 1-65, lower left panel; clone 3-2, lower right panel). Black arrows indicate endothelial cells, white arrows indicate pericytes, and gray arrows indicate acellular capillaries. Figure 2BA comparison of the ratio of endothelial cells to pericytes ("E / P ratio") observed throughout the analyzed retinal area of ​​mice in different treatment groups is shown. To determine the E / P ratio, the number of endothelial cells and pericytes in each captured image was counted. Data are presented as mean ± SD. "***" above the bar indicates a statistically significant difference compared to the IgG treatment group (p < 0.001).

[0038] Figure 2C Comparison of the number of acellular capillaries observed in the entire analyzed retinal area of ​​mice in different treatment groups is shown. Data are presented as mean ± SD. "*" above the bar indicates a statistically significant difference compared to the IgG treatment group (p < 0.05).

[0039] Figure 2D Comparison of observed cell-free capillary lengths is shown. Data are presented as mean ± SD. "**" and "***" above the bars indicate statistically significant differences compared to the IgG-treated group (p < 0.01 and p < 0.001, respectively).

[0040] Figure 3 The therapeutic effect of anti-FAM19A5 antibody on intraretinal vascular congestion in 25-week-old db / db mice using hematoxylin and eosin (H&E) staining is shown. Mice were treated with human IgG control antibody ("IgG", upper right panel) or anti-FAM19A5 antibody (clone "1-65", lower left panel, or clone "3-2", lower right panel). Normal (i.e., non-diabetic) healthy mice (upper left panel) were used as controls. Arrows indicate examples of vascular congestion. The different retinal layers shown include: (i) ganglion cell layer (GCL), (ii) inner plexiform layer (IPL), (iii) inner nuclear layer (INL), (iv) outer nuclear layer (ONL), and (v) retinal pigment epithelium (PE).

[0041] Figure 4 The retinal cell protection of 25-week-old db / db mice treated with anti-FAM19A5 antibodies is shown using TUNEL assay. Mice were treated with human IgG control antibody ("IgG") or anti-FAM19A5 antibody (clone "1-65" or "3-2"). Normal (i.e., non-diabetic) healthy mice were used as controls. The red arrows below the middle column ("TUNEL") indicate that retinal cells are undergoing apoptosis. The intensity of TUNEL staining is related to the degree of apoptosis (e.g., a decrease in intensity indicates partial apoptosis). The different retinal layers shown include: (i) ganglion cell layer (GCL), (ii) inner nuclear layer (INL), (iii) outer nuclear layer (ONL), and (iv) retinal pigment epithelium (PE).

[0042] Figure 5 The inhibitory effect of anti-FAM19A5 antibody on neovascularization in the retina of 25-week-old db / db mice is shown, using immunohistochemical staining to detect CD31 and VEGF expression. Mice were treated with human IgG control antibody ("IgG") or anti-FAM19A5 antibody (clone "1-65" or "3-2"). Normal (i.e., non-diabetic) healthy mice were used as controls. Arrows indicate positive CD31 (top row) or positive VEGF (bottom row) staining.

[0043] Fig. 6A A picture of the image-guided laser system used in Example 8 is shown. Figure 6B Shows exposure to Fig. 6A An exemplary image of the retina being illuminated by laser light produced by an image-guided laser system is shown.

[0044] Fig. 7A and 7B Shown is the inhibitory effect of anti-FAM19A5 antibody (clone 3-2) on choroidal neovascularization (CNV) using fluorescein fundus angiography (FFA) analysis. CNV-induced mice that received human IgG antibody served as controls. Fig. 7A Representative retinal images are shown, showing four wet AMD-like lesions (indicated by arrows in the left column of images) from mice treated with human IgG antibody (top row) or anti-FAM19A5 (clone 3-2) (bottom row) on the day of CNV induction (“Day 0”) and day 13 after CNV induction. Figure 7B Comparison of corrected total fluorescence (CTF) values ​​of CNV-induced mice treated with human IgG antibody ("IgG") or anti-FAM19A5 antibody (clone "3-2") on day 13 after CNV induction is provided. Data are presented as both mean and individual values. "*" indicates a statistically significant difference (p<0.05) compared to the IgG-treated group.

[0045] Fig. 8A , 8B Figures 8A and 8C show the inhibitory effect of anti-FAM19A5 antibody (clone 3-2) on choroidal neovascularization (CNV) using optical coherence tomography (OCT) analysis. CNV-induced mice that received human IgG antibody served as controls. Fig. 8A Representative retinal images are provided, showing four wet AMD-like lesions (numbered 1, 2, 3, and 4) at day 13 after CNV induction in mice treated with human IgG antibody (top row) or anti-FAM19A5 (clone 3-2) (bottom row). The columns labeled 1, 2, 3, and 4 provide OCT images of the corresponding lesions shown in the leftmost image. Figure 8B Scale bars used to quantify lesion size are shown. Figure 8CComparison of CNV lesion size observed on day 13 after CNV induction in mice treated with human IgG control antibody or anti-FAM19A5 antibody (clone 3-2) is provided. Data are presented as both mean and individual values. "***" indicates a statistically significant difference (p<0.001) compared to the IgG treatment group.

[0046] Fig. 9 The effect of anti-FAM19A5 antibody on the recovery of B-wave values ​​in CNV-induced mice is shown. CNV-induced mice were treated with human IgG control antibody ("IgG") or anti-FAM19A5 antibody (clone "3-2"). Normal (i.e., non-diabetic) mice were used as controls. Data are presented as mean ± SD. "*" indicates a statistically significant difference (p < 0.05) compared to the IgG-treated group.

[0047] Fig. 10A and 10B Shown is the inhibitory effect of anti-FAM19A5 antibody (clone 3-2) on choroidal neovascularization (CNV). Fig. 10A Different retinal layers were identified using H&E analysis (left) or OCT analysis (right). The retinal layers shown include: (i) retinal nerve fiber layer / ganglion cell layer (RNFL / GCL); (ii) inner plexiform layer; (iii) inner nuclear layer; (iv) outer plexiform layer; (v) outer nuclear layer, (vi) photoreceptor inner / outer segments (IS / OS), and (vii) retinal pigment epithelium (RPE). Fig. 10B Representative H&E images of relevant cell layers of the retina of different groups of mice are shown: (i) normal healthy animals, (ii) CNV-induced mice treated with human IgG antibody, and (iii) CNV-induced mice treated with anti-FAM19A5 antibody. Arrows indicate choroidal neovascularization areas.

[0048] Fig.11 The inhibitory effect of anti-FAM19A5 antibody on PDGF expression in the inner nerve fiber layer and ganglion cell layer of the retina using IHC analysis is shown. CNV-induced mice were treated with human IgG control antibody ("IgG") or anti-FAM19A5 antibody (clone "3-2"). Normal (i.e., non-diabetic) mice were used as controls. Arrows indicate PDGF-positive staining.

[0049] Fig.12The inhibitory effect of anti-FAM19A5 antibody on oligomeric Aβ and drusen formation in APP / PS-1 / tau transgenic mice (dementia model) using immunofluorescence microscopy is shown. APP / PS-1 / tau transgenic mice have not received (middle row) or have received anti-FAM19A5 antibody treatment (bottom row). Wild-type C57BL / 6 blank mice were used as negative controls (top row). The column labeled "H33342" (nucleic acid staining) shows cells in retinal tissue. The column labeled "QD525-oAβ" shows oligomeric Aβ. The column labeled "647Alexa Phalloidin" shows the formation of drusen, and exemplary drusen are indicated by arrows. The column labeled "Merge" shows the merged images of each treatment group in the above three columns.

[0050] Figures 13A-13C A comparison of the retinal microvascular protective effects of anti-FAM19A5 antibody (clone 1-30) following intravitreal or intravenous injection in a mouse model of diabetic retinopathy is shown. Mice were injected with one of the following: (i) human IgG control antibody ("human IgG"), (ii) aflibercept, (iii) intravitreal anti-FAM19A5 antibody ("IVT"), or (iv) intravenous anti-FAM19A5 antibody ("IV"). Normal (i.e., non-diabetic) healthy mice served as controls. Fig.13A Provided are microscopic images (magnification 100×) of representative retinal areas of mice in various treatment groups after periodic acid-Schiff (PAS) staining. Black arrows indicate endothelial cells, white arrows indicate pericytes, and black arrows indicate acellular capillaries. Fig. 13B A comparison of the number of acellular capillaries observed throughout the analyzed retinal area in mice from different treatment groups is provided. Fig. 13C A comparison of the endothelial cell to pericyte ratio ("E / P ratio") observed throughout the analyzed retinal area of ​​mice in the different treatment groups is provided. To determine the E / P ratio, the number of endothelial cells and pericytes in each captured image was counted. Fig. 13B and 13C Data are presented as mean ± SD. “*” and “***” above the bars indicate statistically significant differences compared with the blank mice (p<0.05 and p<0.001, respectively). “##” and “###” above the bars indicate statistically significant differences compared with the human IgG antibody treatment group (p<0.01 and p<0.001, respectively). Indicates a statistically significant difference compared with the aflibercept treatment group (p<0.001). The “§” above the bar indicates a statistically significant difference compared with the IVT group (p<0.05)

[0051] Fig.14 The results show that anti-FAM19A5 antibody inhibits the formation of intraretinal vascular congestion in diabetic retinopathy mice after intravenous and intravitreal administration. Figures 13A-13C The same as described. Arrows indicate examples of vascular congestion.

[0052] Fig.15 The retinal cell protective effect of anti-FAM19A5 antibody when injected intravitreally or intravenously is shown as measured by TUNEL method. Figures 13A-13C The arrows below the middle row ("TUNEL") indicate that retinal cells are undergoing apoptosis. The intensity of TUNEL staining is correlated with the degree of apoptosis (eg, decreased intensity indicates partial apoptosis).

[0053] Fig.16A and 16B The results show that anti-FAM19A5 antibody inhibits retinal neovascularization in diabetic retinopathy mice after intravitreal and intravenous administration. Figures 13A-13C Same as described above. Using VEGF ( Fig.16A ) and CD31( Fig. 16B ) expression to measure neovascularization. Arrows indicate VEGF or CD31 positive staining. DETAILED DESCRIPTION

[0054] In order to facilitate understanding of the disclosure disclosed herein, a number of terms and phrases are defined. Additional definitions are set forth throughout the detailed description.

[0055] I. Definitions

[0056] In the present disclosure, "an" entity refers to one or more of the entity; for example, "an antibody" can be understood to represent one or more antibodies. Therefore, in this article, "one" (or "a"), "one or more" and "at least one" can be used interchangeably.

[0057] In addition, when "and / or" is used in this document, it should be regarded as a specific disclosure of each of the two specified features or components (with or without the other). Therefore, the term "and / or" used in phrases such as "A and / or B" in this document is intended to include "A and B", "A or B", "A" (alone), and "B" (alone). Similarly, the term "and / or" used in phrases such as "A, B, and / or C" is intended to cover 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).

[0058] It should be understood that whatever aspects are described herein using the language "comprising," similar aspects described within the terms "consisting of" and / or "consisting essentially of" are also provided.

[0059] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art in the relevant field of the present disclosure. For example, Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd edition, 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd edition, 1999, Academic Press; Oxford Dictionary of Biochemistry and Molecular Biology, Revised edition, 2000, Oxford University Press, provide a general dictionary of many terms used in the present disclosure for technicians.

[0060] Units, prefixes and symbols are expressed in the form accepted by the International System of Units (SI). Numerical ranges include numbers that limit the range. Unless otherwise indicated, amino acid sequences are written from left to right in the direction of amino to carboxyl. The headings provided herein are not limitations on the various aspects of the disclosure and can be obtained by reference to the specification as a whole. Therefore, by reference to the specification, the terms defined below are more fully defined as a whole.

[0061] As used herein, the term "about" is used to mean approximately, roughly, around, or in the region of. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values ​​recited. The term "about" generally modifies a numerical value above and below the stated value by a variable up or down (increase or decrease), such as 10%.

[0062] As used herein, the term "retinal disease" refers to disease, inflammation, or damage to the retina (ie, the tissue lining the inner surface of the back of the eye that captures images of the body) that passes through the cornea and lens.

[0063] The term "diabetic retinopathy" (DR) as used herein refers to retinopathy caused by complications of diabetes. Depending on the severity of the disease, DR may be asymptomatic, cause mild vision problems, or lead to blindness. DR is caused by changes in retinal microvessels. Death of pericytes in the wall and thickening of the basement membrane caused by high blood sugar can cause vascular wall dysfunction. These injuries can alter the formation of the blood-retinal barrier and increase the permeability of retinal blood vessels. When high blood sugar continues to activate protein kinase C-δ (PKC-δ, encoded by Prkcd) and p38 mitogen-activated protein kinase (MAPK) to increase the expression of PKC-δ signaling, phosphatase-1 (SHP-1) containing the Src homology region 2 domain, and previously unknown targets of protein tyrosine phosphatases, pericyte death can be caused. This signaling cascade can lead to dephosphorylation of the PDGF receptor and a decrease in downstream signaling from the receptor, leading to apoptosis of pericytes. Small blood vessels (such as those in the eye) are particularly susceptible to poor glycemic control. Excessive accumulation of glucose and / or fructose can damage the tiny blood vessels in the retina.

[0064] DR can be divided into two separate stages. Wu L. et al., World J Diabetes 4(6):290-294 (2013). The first stage is called non-proliferative diabetic retinopathy (NPDR) and is associated with early diabetic retinopathy. NPDR is generally asymptomatic or accompanied by mild visual distortion (caused by fluid leakage into the tissue surrounding blood vessels). The only way to detect NPDR is through fundus photography, which can reveal microaneurysms (tiny bulges of blood filled in the artery wall). If left untreated, DR patients can progress to the second and more advanced stage, called proliferative diabetic retinopathy (PDR). PDR is characterized by abnormal formation of new blood vessels (i.e., neovascularization), which can rupture and bleed, causing blurred vision. Other symptoms of PDR include, but are not limited to: dark spots or lines floating in the field of vision ("floaters"), fluctuations in vision, impaired color vision, dark or blank areas in the field of vision, pain, patchy visual fields, and complete vision loss.

[0065] 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 maculopathy, and diabetic macular edema.

[0066] In some aspects, PDR occurs after the onset of NPRD (e.g., an individual is first diagnosed with NPRD and then develops PDR). In other aspects, the occurrence of PDR is independent of NPRD. As used herein, the term "diabetic retinopathy" also includes all types of diabetic retinopathy, regardless of the cause of diabetic retinopathy and all its symptoms. Non-limiting examples of risk factors for diabetic retinopathy include: duration of diabetes, heredity, excessive drinking, smoking, hypertension, obesity, dyslipidemia, hypercholesterolemia, kidney disease, pregnancy, and kidney damage.

[0067] As used herein, the term "maculopathy" refers to any pathological condition of the macula (i.e., the area in the center of the retina associated with highly sensitive, highly accurate vision). In some aspects, the terms "maculopathy" and "retinopathy" can be used interchangeably (i.e., only the macula is affected). In some aspects, the maculopathy is a diabetic maculopathy.

[0068] "Diabetic maculopathy" occurs when the macula is affected by retinal changes caused by diabetes. The term refers to two different eye diseases: diabetic macular edema and diabetic ischemic maculopathy. The two types of maculopathy often coexist, that is, patients with macular edema often also have ischemic maculopathy. Even mild macular edema can occur with ischemic maculopathy and macular edema. In certain aspects, retinal changes associated with diabetic maculopathy in an individual's retina include: decreased retinal potential, loss of pericytes, formation of acellular capillaries, vascular congestion, impaired vascular function, vascular leakage, vascular occlusion, tissue swelling (edema), tissue hypoxia, or any combination thereof.

[0069] As used herein, the term "acellular capillaries" refers to capillary-sized blood vessels that lack cell nuclei anywhere along their length.

[0070] As used herein, the term "vascular congestion" refers to a type of vascular damage that is an important factor in the pathogenesis of various eye conditions disclosed herein (e.g., diabetic macular edema). Vascular congestion is associated with the accumulation of fluid within vascular tissue (e.g., blood within blood vessels). In certain aspects, vascular congestion can be caused by hyperglycemia (i.e., high blood sugar).

[0071] As used herein, the term "macular degeneration" refers to any number of diseases and conditions in which the macula degenerates or loses functional activity. For example, the degeneration or loss of functional activity can be caused by cell death, cell proliferation reduction, loss of normal biological function or the above combination. Macular degeneration can cause and / or be manifested as changes in the structural integrity of macular cells and / or extracellular matrix, changes in normal cells and / or extracellular matrix construction and / or loss of macular cell function. The cell can be any cell type that is usually present in or near the macula, including RPE cells, photoreceptors and / or capillary endothelial cells. Age-related macular degeneration is the most common macular degeneration, but the term "macular degeneration" does not necessarily exclude macular degeneration in non-elderly patients. Non-limiting examples of macular degeneration include: age-related macular degeneration (wet or dry); Best macular dystrophy, Sorsby fundus dystrophy, Mallatia Leventinese, Doyne honeycomb retinal dystrophy, Stargardt's disease (also known as Stargardt macular dystrophy, juvenile macular degeneration or fundus yellow spots), and pigment epithelial detachment-related macular degeneration.

[0072] As used herein, term "age-related macular degeneration" (AMD) refers to a retinopathy that is usually influential in the elderly, is relevant to central vision loss, and is caused by damage to the central part of the retina (i.e., macula). Generally, AMD is characterized by the gradual accumulation or aggregation of yellow insoluble extracellular deposits in the macula (mainly between the retinal pigment epithelium (RPE) and the underlying choroid), known as drusen (accumulation of extracellular proteins, such as beta amyloid and lipids). The accumulation or aggregation of these deposits in the macula can cause the macula to gradually degenerate, thereby causing central vision damage. As used herein, term "macula" refers to the central part of the retina responsible for central high-resolution color vision.

[0073] The pathogenesis of age-related macular degeneration is currently unclear, although several theories have been proposed, including oxidative stress, mitochondrial dysfunction, and inflammatory processes. An imbalance between the production and degradation of damaged cellular components leads to the accumulation of harmful products, such as intracellular lipofuscin and extracellular drusen. Initial atrophy is marked by areas of thinning or depigmentation of the retinal pigment epithelium (RPE) (preceding geographic atrophy in the early stages of AMD). In late AMD, RPE atrophy (geographic atrophy) and / or neovascularization develops, leading to photoreceptor death and loss of central vision. In dry (non-exudative) AMD, cellular debris called drusen accumulate between the retina and the choroid, resulting in retinal atrophy and scarring. In the more severe wet (exudative) AMD, blood vessels grow from the choroid behind the retina (neovascularization), which can lead to exudate and fluid leakage, and can also cause bleeding.

[0074] Depending on the extent of existing drusen, AMD can be divided into three main stages: (i) early, (ii) intermediate and (iii) late. Early AMD is characterized by the presence of several small (e.g., less than about 63 microns in diameter) drusen, or the presence of several medium-sized (e.g., between about 63 and 124 microns in diameter). Patients have no obvious symptoms and vision loss in the early stages. Intermediate stage is characterized by the presence of many medium-sized drusen, or one or more large (e.g., greater than about 125 microns in diameter). At this stage, some patients may begin to experience blurred spots in the center of their visual field. Late AMD is characterized by extensive damage to retinal tissue, resulting in a central blind spot and ultimately loss of central vision. Late AMD can be further divided into two isotypes based on the type of damage (e.g., the presence or absence of neovascularization): (i) geographic atrophy (also known as atrophic AMD) and (ii) wet AMD (also known as neovascular or exudative AMD).

[0075] AMD has two main forms: (i) dry AMD and (ii) wet AMD. Unless otherwise specified, the term "age-related macular degeneration" includes dry AMD and wet AMD. The term "age-related macular degeneration" used herein also includes all types of age-related macular degeneration, regardless of the cause of age-related macular degeneration and all symptoms thereof. Non-limiting examples of macular degeneration (such as age-related macular degeneration) related symptoms include: central vision loss, distortion, reduced contrast sensitivity, blurred vision, difficulty adapting to dim light, sudden illness and rapid deterioration of symptoms and decreased color vision. In some aspects, macular degeneration (such as age-related macular degeneration) can cause macular edema (i.e., fluid and protein deposits above or below the macula, causing swelling of the macula).

[0076] The term "dry AMD" (also called atrophic age-related macular degeneration or non-exudative AMD) used herein refers to all forms of AMD that are not wet (neovascular) AMD. These include early and intermediate forms of AMD, as well as late dry AMD, known as geographic atrophy. Patients with dry AMD tend to have mild symptoms early on; if the disease progresses to geographic atrophy, the incidence of visual function loss is higher.

[0077] Term used herein " wet AMD " (also referred to as neovascular age-related macular degeneration or exudative AMD) refers to the retinal state characterized by the presence of retinal neovascularization, which is the most advanced form of AMD. In wet AMD, blood vessels grow out of the choroidal capillaries through the defects of Bruch's membrane, in some cases, through the lower retinal pigment epithelium (choroidal neovascularization or angiogenesis). The tissue of the serous or hemorrhagic exudates flowing out of these blood vessels can cause the formation of fibrovascular scars in the macula, and is accompanied by detachment and tearing, vitreous hemorrhage and permanent central vision loss of neuroretinal degeneration, retinal pigment epithelium.

[0078] As used herein, the term "neovascularization" refers to the growth of new abnormal blood vessels in different parts of the eye, which can cause hemorrhage and vision loss. As used herein, the term "choroidal neovascularization" refers to the abnormal growth of neovascularization in the choroid (i.e., the eye vascular layer containing connective tissue between the retina and the sclera). In wet AMD, neovascularization can grow through the retinal pigment epithelium (RPE) and choroid, enter the retina, and due to blood and lipid leakage, visual function can be damaged. As used herein, the term "retinal neovascularization" refers to the abnormal development, proliferation and / or growth of blood vessels on or in the retina (such as the retinal surface) of the retina. Retinal neovascularization can develop in many retinal diseases, such as retinopathy associated with retinal ischemia and / or inflammatory diseases (e.g., diabetic retinopathy, sickle cell retinopathy, Eales' disease, ocular ischemic syndrome, carotid cavernous fistula, familial exudative vitreoretinopathy, hyperviscosity syndrome, idiopathic occlusive arteritis, radiation retinopathy, retinal vein occlusion, retinal artery occlusion, retinal embolism, scattered retinochoroidopathy, retinal vasculitis, sarcoidosis, toxoplasmosis, uveitis, choroidal melanoma, chronic retinal detachment, anterior ischemic optic neuropathy (AION), non-arteritic anterior ischemic optic neuropathy (NAION), and pigment disorder). Neovascularization detection methods known in the art include, but are not limited to, detecting CD31 (also known as platelet endothelial cell adhesion molecule, PECAM-1) and vascular endothelial growth factor (VEGF) expression in tissues. For example, see Schluter A. et al., BMC Cancer 18(1): 272 (2018).

[0079] The retinal pigment epithelium (RPE) is a single layer of epithelial cells located at the back of the vertebrate eye between the choroidal blood supply (choriocapillaris) and the retinal neural layer. The RPE is one component of the blood-retinal barrier and RPE cells play an important role in maintaining the visual cycle, phagocytosis of photoreceptor outer segments, and transport of nutrients, metabolic waste products, ions, and fluid between the distal retina and the choriocapillaris.

[0080] The term "intraocular" refers to the inside or below of ocular tissue. As used herein, the term "intraocular administration" refers to any administration that can deliver a composition (e.g., an anti-FAM19A5 antibody disclosed herein) to the subfascial, subconjunctival, suprachoroidal, intravitreal, and other sites of the eye. In some aspects, intraocular administration includes intravitreal administration.

[0081] The term "ocular condition" refers to a disease, nutrient or condition that affects or involves the eye or a part or region of the eye, such as a retinal disease. In certain aspects, ocular conditions that can be treated by the present disclosure include diabetic retinopathy and age-related macular degeneration. Other ocular conditions that can be treated by the compositions and methods disclosed herein include diabetic macular edema, anterior ischemic optic neuropathy (AION), non-arteritic anterior ischemic optic neuropathy (NAION), retinal vein occlusion, retinal artery occlusion, or a combination thereof.

[0082] The term "eye" as used herein includes the eyeball and the tissues and fluids that make up the eyeball (such as the sclera, cornea, anterior and posterior eye chambers, iris / pupil, lens, vitreous body, retina, fovea, macula and choroid), the muscles around the eye (such as the oblique muscles and rectus muscles), and the portion of the optic nerve in or near the eyeball.

[0083] The term "optic nerve damage" refers to changes in the normal structure or function of the optic nerve. Changes in the normal structure or function of the optic nerve may be caused by any disease, disorder or injury, including glaucoma. Changes in the normal function of the optic nerve include any changes in the ability of the optic nerve to work properly, such as transmitting visual information from the retina to the brain. Functional changes can manifest, such as loss of visual field, impaired central vision, abnormal color vision, etc. Examples of structural changes include loss of nerve fibers in the retina, abnormal optic cup enlargement of the optic nerve, and / or loss of retinal ganglion cell layer cells. "Optic nerve damage" as used herein may include optic nerve damage to one or both optic nerves of an individual.

[0084] The terms "treat", "treating" and "treatment" as used herein refer to any type of intervention or process performed on an individual or to an individual with an active agent, with the purpose of reversing, alleviating, improving, inhibiting or slowing down or preventing the progression, development, severity or recurrence of symptoms, complications, conditions or biochemical indicators associated with a disease. Treatment can be treatment of an individual with a disease or an individual without a disease (e.g., for prevention).

[0085] The term "administered" as used herein refers to the use of any of the various methods and delivery systems known to those skilled in the art to physically introduce a therapeutic agent or a composition comprising a therapeutic agent into an individual. The different routes of administration of the antibodies described herein include intravenous, intraperitoneal, intramuscular, subcutaneous, spinal or other parenteral routes of administration, such as by injection or infusion. The phrase "parenteral administration" as used herein refers to a mode of administration other than enteral and topical administration, usually by injection, including but not limited to intravenous, intraperitoneal, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, transtracheal, intratracheal, pulmonary, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraventricular, intravitreal, epidural and intrasternal injection and infusion, and in vivo electroporation. The antibodies described herein may also be administered via non-parenteral routes, such as local, epidermal or mucosal routes of administration, such as intranasal, oral, vaginal, rectal, sublingual or topical administration. For example, administration may also be performed once, multiple times and / or over one or more extended time periods.

[0086] As used herein, the term "therapeutically effective amount" refers to an amount of a drug, alone or in combination with another therapeutic agent, that is used to effectively "treat" an individual's disease or condition or to reduce the risk, potential, likelihood, or occurrence of a disease or condition (e.g., glaucoma). A "therapeutically effective amount" includes an amount of a drug or therapeutic agent that provides some improvement or benefit to an individual suffering from a disease or condition (e.g., glaucoma disclosed herein) or an individual at risk of suffering from the disease or condition. Thus, a "therapeutically effective" amount is an amount that reduces the risk, potential, likelihood, or occurrence of a disease, or provides a condition or some alleviation, alleviation, and / or reduction of at least one indicator (e.g., increased inflammation) and / or reduces at least one clinical symptom of a disease or condition.

[0087] The term "individual" as used herein includes any human or non-human animal. The term "non-human animal" includes all vertebrates, such as mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc.

[0088] The term "sequence similarity family 19 member A5" or "FAM19A5" refers to a protein belonging to the TAFA family (also known as the FAM19 family) of five highly homologous proteins. Tang TY et al., Genomics 83(4):727-34 (2004). These proteins contain conserved cysteine ​​residues at fixed positions and are distantly related to macrophage inflammatory protein 1-alpha (MIP-l-alpha), a member of the CC-chemokine family. TAFA proteins are expressed in specific regions of the brain and spinal cord. Studies have shown that these proteins are produced and secreted by adult neural stem cells during neurogenesis. FAM19A5 is also known as TAFA5 or chemokine-like protein TAFA-5.

[0089] It is believed that FAM19A5 is of great significance for the development, differentiation and formation of the complete central nervous system. FAM19A5 also plays an important role in the pathogenesis of many central nervous system injuries and / or degenerative brain diseases (such as Huntington's disease, Parkinson's disease, Alzheimer's disease, cerebral spinal cord injury, stroke and brain tumors). After central nervous system damage, neural stem cells produce FAM19A5, which induces normal astrocytes to differentiate into reactive astrocytes. These reactive astrocytes (together with microglial cells) can express a variety of ECMs (such as proteoglycans) and induce glial scar formation, which surrounds the damaged area of ​​the central nervous system in a reticular form, preventing neuronal regeneration. Antagonists for FAM19A5 (such as anti-FAM19A5 antibodies) can be used to prevent and / or treat central nervous system damage and / or diseases. Referring to U.S. Patent No. 9,579,398.

[0090] However, for some diseases, especially those associated with vascular abnormalities, a recent study suggests that FAM19A5 antagonists may not be a viable treatment option. Wang et al., Circulation 138(l): 48-63 (2018). Wang et al. showed that adipose tissue of normal healthy people produces FAM19A5, which signals through sphingosine-l-phosphate receptor 2 (S1PR2), inhibits the proliferation and migration of vascular smooth muscle cells (VSMCs), and reduces neointimal formation after injury. However, in obese individuals, FAM19A5 expression (mRNA and protein) in adipose tissue is significantly reduced. Wang et al. concluded that reduced FAM19A5 activity is responsible for the accelerated development of cardiovascular disease in many obese individuals.

[0091] The human genetically encoded FAM19A5 is located on chromosome 22. There are three human FAM19A5 (UniProt: Q7Z5A7) isoforms, which are thought to be produced by alternative splicing: isoform 1 (UniProt: Q7Z5A7-1), consisting of 132 amino acids; isoform 2 (UniProt: Q7Z5A7-2), consisting of 125 amino acids; isoform 3 (UniProt: Q7Z5A7-3), consisting of 53 amino acids. The human FAM19A5 protein is thought to exist in membrane-bound and soluble (secreted) forms. Isoform 1 is thought to be a membrane protein with one transmembrane region. Isoform 2, which was reported as a secreted protein (soluble) in Tang TY et al., Genomics 83 (4): 727-34 (2004), contains a signal peptide at amino acids 1-25. Isoform 3 is predicted based on EST data. Below are the amino acid sequences of the three known human FAM19A5 isoforms.

[0092] (I) Isoform 1 (UniProt: Q7Z5A7-1, transmembrane protein): This isoform was selected as the canonical sequence.

[0093] MAPSPRTGSR QDATALPSMS STFWAFMILA SLLIAYCSQL AAGTCEIVTL DRDSSQPRRTIARQTARCAC RKGQIAGTTR ARPACVDARI IKTKQWCDML PCLEGEGCDL LINRSGWTCT QPGGRIKTTTVS(SEQ ID NO: 1)

[0094] (II) Isoform 2 (UniProt: Q7Z5A7-2, soluble protein):

[0095] MQLLKALWAL AGAALCCFLV LVIHAQFLKE GQLAAGTCEI VTLDRDSSQP RRTIARQTARCACRKGQIAG TTRARPACVD ARIIKTKQWC DMLPCLEGEG CDLLINRSGW TCTQPGGRIK TTTVS(SEQID NO: 2)

[0096] (III) Isoform 3 (UniProt: Q7Z5A7-3):

[0097] MYHHREWPAR IIKTKQWCDM LPCLEGEGCD LLINRSGWTC TQPGGRIKTT TVS(SEQ ID NO: 3)

[0098] The term "FAM19A5" includes any variant or isoform of FAM19A5 naturally expressed by cells. Therefore, the antibodies described herein may cross-react with different isoforms in the same species (e.g., different isoforms of human FAM19A5), or cross-react with FAM19A5 from non-human species (e.g., mouse FAM19A5). Alternatively, the antibody may be specific to human FAM19A5 and may not exhibit any cross-reactivity with other species. FAM19A5 or any of its variants and isoforms may be isolated from cells or tissues that naturally express them, or recombinantly produced. The polynucleotide encoding human FAM19A5 has GenBank accession number BC039396 and the following sequence:

[0099] Table 1A. Polynucleotide sequence of human FAM19A5

[0100]

[0101] The term "antagonist against FAM19A5 protein" refers to all antagonists that inhibit and / or neutralize the expression and / or activity of FAM19A5 protein. Such antagonists can be peptides, nucleic acids or compounds. More specifically, the antagonists can be antisense oligonucleotides, siRNA, shRNA, miRNA, dsRNA, aptamers, PNA (peptide nucleic acid) targeting FAM19A5 or vectors comprising them. In certain aspects, the antagonist can be an antibody or an antigen binding portion thereof that specifically binds to the FAM19A5 protein.

[0102] The terms "antibody and antibodies" are terms of the art and are used interchangeably herein to refer to molecules having an antigen binding site that specifically binds to an antigen. The terms used herein include complete antibodies and any antigen binding fragments thereof (i.e., "antigen binding portions") or single chains. In one aspect, an "antibody" refers to a glycoprotein or an antigen binding portion thereof comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. On the other hand, an "antibody" refers to a single-chain antibody comprising a single variable domain (e.g., a VHH domain). Each heavy chain consists of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. In some naturally occurring antibodies, the heavy chain constant region consists of three domains, CH1, CH2, and CH3. In some naturally occurring antibodies, each light chain consists of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region consists of one domain, CL.

[0103] The VH and VL regions can be further subdivided into hypervariable regions called complementarity determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant region of an antibody can mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0104] The term "Kabat numbering" and similar terms are art-recognized terms that refer to a system for numbering amino acid residues in the heavy and light chain variable regions of an antibody or its antigen-binding portion. In certain aspects, the CDRs of an antibody can be determined according to the Kabat numbering system (e.g., see Kabat EA and Wu TT (1971) Ann NY Acad Sci 190: 382-391 and Kabat EA et al., (1991) Sequences of Proteins of Immunological Interest, 5th edition, U.S. Department of Health and Human Services, NIH Publication No.: 91-3242). Using the Kabat numbering system, the CDRs within an antibody heavy chain molecule are typically present at amino acids 31 to 35 (which may optionally include one or two additional amino acids), (after position 35 (referred to as 35A and 35B in the Kabat numbering scheme) (CDR1), amino acids 50 to 65 (CDR2), and amino acids 95 to 102 (CDR3). Using the Kabat numbering system, the CDRs within an antibody light chain molecule are typically present at amino acids 24 to 34 (CDR1), amino acids 50 to 56 (CDR2), and amino acids 89 to 97 (CDR3). In a specific aspect, the antibody CDRs described herein have been determined according to the Kabat numbering scheme.

[0105] The phrases "amino acid position numbering as in Kabat", "Kabat position" and grammatical variants thereof refer to the numbering system for heavy chain variable domains or light chain variable domains compiled for antibodies in Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, Public Health Service, National Institutes of Health, Bethesda, Maryland (1991). Using this numbering system, the actual linear amino acid sequence may contain fewer or more amino acids, corresponding to shortening of the variable domain FW or CDR, or to insertion into the FW or CDR. For example, the heavy chain variable domain may include a single amino acid insertion after residue 52 of H2 (residue 52a according to Kabat) and an inserted residue after residue 82 of the heavy chain FW (e.g., residues 82a, 82b, and 82c, etc. according to Kabat). See Table 1B.

[0106] Table 1B

[0107]

[0108] For a given antibody, the Kabat numbering of residues can be determined by aligning the homologous region of the antibody sequence with the "standard" Kabat numbering sequence. Chothia refers to the location of the structural loops (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). When numbering using the Kabat numbering convention, the ends of the Chothia CDR-H1 loop vary between H32 and H34, depending on the length of the loop (because the Kabat numbering scheme places the insertion at H35A and H35B; if both 35A and 35B are absent, the loop ends at 32; if only 35A is present, the loop ends at 33; if both 35A and 35B are present, the loop ends at 34). The AbM hypervariable regions represent a compromise between the Kabat CDRs and the Chothia structural loops and are employed by Oxford Molecular's AbM antibody modeling software.

[0109] IMGT (ImMunoGeneTics) also provides a numbering system for immunoglobulin variable regions (including CDRs). See, for example, Lefranc, MP et al., Dev. Comp. Immunol. 27:55-77 (2003), which is incorporated herein by reference. The IMGT numbering system is based on alignments of more than 5000 sequences, structural data, and characterization of hypervariable loops, making it easy to compare variable and CDR regions of all species. According to the IMGT numbering scheme, VH-CDR1 is at positions 26 to 35, VH-CDR2 is at positions 51 to 57, VH-CDR3 is at positions 93 to 102, VL-CDR1 is at positions 27 to 32, VL-CDR2 is at positions 50 to 52, and VL-CDR3 is at positions 89 to 97.

[0110] For all heavy chain constant region amino acid positions discussed in this disclosure, numbering is according to the EU index first described in Edelman et al. (1969) Proc. Natl. Acad. Sci. 63(1):78-85, which describes the amino acid sequence of the myeloma protein EEG, the first human IgG1 sequenced. The EU index of Edelman et al. is also set forth in Kabat et al. (1991) Sequences of Proteins of Immunological Interest, 5th edition (Public Health Service, National Institutes of Health, Bethesda, Maryland). Thus, the phrases "EU index as set forth in Kabat" or "EU index of Kabat" and "position ... according to the EU index as set forth in Kabat" and grammatical variations thereof refer to the residue numbering system for the human IgG1 EU antibody of Edelman et al. as set forth in Kabat 1991.

[0111] Kabat 1991 proposed a numbering system for amino acid sequences of variable domains (heavy and light chains) and light chain constant regions.

[0112] The antibody can be an immunoglobulin molecule of any type (e.g., IgG, IgE, IgM, IgD, IgA or IgY), any class (e.g., IgD, IgG2, IgG3, IgG4, IgA1 or IgA2) or any subclass (e.g., human IgG1, IgG2, IgG3 and IgG4; and mouse IgG1, IgG2a, IgG2b and IgG3). Immunoglobulins (e.g., IgG1) exist in several allotypes that differ from each other in at most a few amino acids. The antibodies disclosed herein can be from any commonly known isotype, class, subclass or allotype. In certain aspects, the antibodies described herein are antibodies of the IgG1, IgG2, IgG3 or IgG4 subclasses or any hybrid thereof. In certain aspects, the antibodies are antibodies of the human IgG1 subclass or human IgG2 or human IgG4 subclasses.

[0113] "Antibodies" include, for example, naturally occurring and non-naturally occurring antibodies; monoclonal and polyclonal antibodies; chimeric and humanized antibodies; human and non-human antibodies; fully synthetic antibodies; single chain antibodies; monospecific antibodies; multispecific antibodies (including bispecific antibodies); tetrameric antibodies comprising two heavy chain and two light chain molecules; antibody light chain monomers; antibody heavy chain monomers; antibody light chain dimers, antibody heavy chain dimers; antibody light chain-antibody heavy chain pairs; intrabodies; heteroconjugate antibodies; monovalent antibodies; camelized antibodies; affibodies; anti-idiotypic (anti-Id) antibodies (including, for example, anti-anti-Id antibodies) and single domain antibodies (sdAbs), which include binding molecules composed of a single monomeric variable antibody domain (e.g., VH domain or VL domain) that is fully capable of antigen binding. Harmen MM and Haard HJ Appl Microbiol Biotechnol 77(1): 13-22 (2007)).

[0114] As used herein, the term "antigen-binding portion" of an antibody refers to one or more fragments of an antibody that retains the ability to specifically bind to an antigen (e.g., human FAM19A5). Such "fragments" are, for example, about 8 to about 1500 amino acids in length, with a suitable length of about 8 to about 745 amino acids, a suitable length of about 8 to about 300 amino acids, such as about 8 to about 200 amino acids, or about 10 to about 50 or 100 amino acids. Facts show that the antigen-binding function of an antibody can be performed by a fragment of a full-length antibody. Examples of binding fragments included within the term "antigen-binding portion" of an antibody (e.g., the anti-FAM19A5 antibodies described herein) include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bond at the hinge region; (iii) a Fd fragment, consisting of the VH and CH1 domains; (iv) a Fv fragment, consisting of the VL and VH domains of a single arm of an antibody, and disulfide-linked Fvs (sdFv); (v) a dAb fragment (Ward et al., (1989) Nature 341:544-546), consisting of a VH domain; and (vi) isolated complementarity determining regions (CDRs) or (vii) a combination of two or more isolated CDRs, wherein the CDRs may optionally be linked by a synthetic linker. In addition, although the two domains of the Fv fragment, VL and VH, are encoded by separate genes, they can be connected by synthetic linkers using recombinant methods to form a single protein chain in which the VL and VH regions are paired to form a monovalent molecule (called single-chain Fv (scFv)); for example, see Bird et al. (1988) Science 242: 423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85: 5879-5883). Such single-chain antibodies are also expected to be included in the term "antigen-binding portion" of the antibody. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened in the same manner as intact antibodies to achieve their utility. The antigen-binding portion can be produced by recombinant DNA technology, or by enzymatic or chemical cleavage of intact immunoglobulins.

[0115] The terms "variable region" or "variable domain" as used herein are interchangeable and are common terms in the art. The variable region generally refers to a portion of an antibody, generally a portion of a light chain or a heavy chain, generally about 110 to 120 amino acids at the amino terminal in a mature heavy chain and about 90 to 115 amino acids in a mature light chain, which vary greatly in sequence between antibodies and are used for the binding and specificity of a particular antibody to its specific antigen. The variability of the sequence is concentrated in regions called complementary determining regions (CDRs), while the more conserved regions in the variable domain are called framework regions (FRs).

[0116] Without wishing to be bound by any particular mechanism or theory, it is believed that the CDRs of the light chain and heavy chain are primarily responsible for the interaction and specificity of the antibody with the antigen. In some aspects, the variable region is a human variable region. In some aspects, the variable region comprises rodent or mouse CDRs and human framework regions (FRs). In specific aspects, the variable region is a primate (e.g., non-human primate) variable region. In some aspects, the variable region comprises rodent or mouse CDRs and primate (e.g., non-human primate) framework regions (FRs).

[0117] As used herein, the term "heavy chain" (HC), when used in reference to antibodies, may refer to any of the different types based on the amino acid sequence of the constant domain, such as α (a), δ (d), ε (e), γ (g), and μ (m), which give rise to IgA, IgD, IgE, IgG, and IgM class antibodies, respectively, including IgG subclasses, such as IgG1, IgG2, IgG3, and IgG4.

[0118] As used herein, the term "light chain" (LC) when used to refer to an antibody can refer to any of the different types based on the amino acid sequence of the constant domain, such as kappa (K) or lambda (I). The light chain amino acid sequence is an amino acid sequence well known in the art. In a specific aspect, the light chain is a human light chain.

[0119] The terms "VL" and "VL domain" are used interchangeably to refer to the light chain variable region of an antibody.

[0120] The terms "VH" and "VH domain" are used interchangeably to refer to the heavy chain variable region of an antibody.

[0121] As used herein, the terms "constant region" or "constant domain" are interchangeable and have their usual meaning in the art. The constant region is the portion of an antibody, such as the carboxyl terminal portion of a light chain and / or a heavy chain, which is not directly involved in binding the antibody to an antigen, but may exhibit various effector functions, such as interacting with Fc receptors. The constant region of an immunoglobulin molecule generally has an amino acid sequence that is more conserved relative to an immunoglobulin variable domain.

[0122] "Fc region" (fragment crystallizable region) or "Fc domain" or "Fc" refers to the C-terminal region of an antibody heavy chain that can mediate the binding of immunoglobulins to host tissues or factors, including binding to Fc receptors located on various cells of the immune system (e.g., effector cells), or to the first component (C1q) of the classical complement system. Thus, the Fc region includes the constant region of the antibody, excluding the first constant region immunoglobulin domain (e.g., CH1 or CL). In IgG, IgA, and IgD antibody isotypes, the Fc region includes two identical protein fragments derived from the second (CH2) and third (CH3) constant domains of the two heavy chains of the antibody; IgM and IgE Fc regions include three heavy chain constant domains (CH domains 2-4) in each polypeptide chain. For IgG, the Fc region includes immunoglobulin domains Cγ2 and Cγ3, and the hinge between Cγ1 and Cγ2. Although the boundaries of the Fc region of an immunoglobulin heavy chain may vary, the Fc region of a human IgG heavy chain is generally defined as extending from an amino acid residue at position C226 or P230 (or an amino acid between these two amino acids) to the carboxyl terminus of the heavy chain, wherein numbering is performed according to the EU index in Kabat. The CH2 domain of the human IgG Fc region extends from about amino acid 231 to about amino acid 340, while the CH3 domain is located on the C-terminal side of the Cm domain in the Fc region, i.e., from about amino acid 341 of IgG to about amino acid 447. The Fc region used herein may be a native sequence Fc, including any allotypic variants, or a variant Fc (e.g., a non-naturally occurring Fc). Fc may also refer to the region in an isolated state or within the context of an Fc protein polypeptide, such as a "binding protein containing an Fc region", also referred to as an "Fc fusion protein" (e.g., an antibody or immunoadhesion).

[0123] A "native sequence Fc region" or "native sequence Fc" includes an amino acid sequence that is identical to the amino acid sequence of a naturally occurring Fc region. Native sequence human Fc regions include native sequence human IgG1 Fc regions; native sequence human IgG2 Fc regions; native sequence human IgG3 Fc regions; and native sequence human IgG4 Fc regions and naturally occurring variants thereof. Native sequence Fc includes various Fc allotypes (e.g., see Jefferis et al. (2009) Monoclonal Antibodies (mAbs) 1:1; Vidarsson G. et al. Frontiers in Immunology (Front Immunol) 5:520 (published online on October 20, 2014)).

[0124] "Fc receptor" or "FcR" is a receptor that binds to the Fc region of an immunoglobulin. FcRs that bind to IgG antibodies include receptors of the FcyR family, including allelic variants and alternative splicing forms of these receptors. The FcγR family consists of three activating receptors (mouse FcγRI, FcγRIII and FcγRIV; human FcγRIA, FcγRIIA and FcγRIIIA) and one inhibitory receptor (FcγRIIB). Human IgG1 binds to most human Fc receptors and elicits the strongest Fc effector function. In terms of the type of activating Fc receptor it binds, it is considered equivalent to mouse IgG2a. In contrast, human IgG4 elicits minimal Fc effector function. Vidarsson G. et al. Frontiers in Immunology 5: 520 (published online on October 20, 2014).

[0125] For example, the constant region can be manipulated by recombinant technology to eliminate one or more effector functions. "Effector function" refers to the interaction of the antibody Fc region with an Fc receptor or ligand, or the biochemical events resulting therefrom. Exemplary "effector functions" include C1q binding, complement-dependent cytotoxicity (CDC), Fc receptor binding, FcγR-mediated effector functions, such as ADCC and antibody-dependent cell-mediated phagocytosis (ADCP) and downregulation of cell surface receptors (e.g., B cell receptor; BCR). Such effector functions generally require the Fc region to be combined with a binding domain (e.g., an antibody variable domain). Therefore, the term "constant region without Fc function" includes a constant region mediated by an Fc region with reduced effector functions or without one or more effector functions.

[0126] The effector function of an antibody can be reduced or avoided by different methods. By using an antibody fragment lacking an Fc region (e.g., Fab, F(ab')2, single-chain Fv (scFv), or sdAb consisting of a monomeric VH or VL domain), the effector function of an antibody can be reduced or avoided. Alternatively, by removing the sugars attached to specific residues in the Fc region, so-called non-glycosylated antibodies can be generated to reduce the effector function of the antibody while retaining other valuable properties of the Fc region (e.g., extended half-life and heterodimerization). For example, glycosylated antibodies can be generated by deleting or changing the residues to which the sugar is attached, enzymatically removing the sugar, producing antibodies in cells cultured in the presence of a glycosylation inhibitor, or expressing antibodies in cells (e.g., bacterial host cells) that cannot glycosylate proteins. For example, see U.S. Patent (Publication No.: 20120100140). Another method is to use an Fc region with reduced effector function from an IgG subclass, for example, IgG2 and IgG4 antibodies are characterized by having a lower level of Fc effector function than IgG1 and IgG3. In the CH2 domain of the Fc portion, the residues closest to the hinge region are responsible for the effector function of the antibody, as this residue contains the most overlapping binding sites for C1q (complement) and IgG-Fc receptors (FcγR) on effector cells of the innate immune system. Vidarsson G. et al. Front Immunol 5:520 (published online on Oct. 20, 2014). Thus, antibodies with reduced or no Fc effector function can be prepared by generating, for example, a chimeric Fc region comprising a CH2 domain from an IgG antibody of the IgG4 isotype and a CH3 domain from an IgG antibody of the IgG1 isotype, or a chimeric Fc region comprising a hinge region from IgG2 and a CH2 region from IgG4 (e.g., see Lau C. et al., J. Immunol. 191: 4769-4777 (2013)), or an Fc region with mutations that result in altered Fc effector function (e.g., reduced or no Fc function). Such Fc regions with mutations are known in the art. For example, see U.S. Patent Publication No. 20120100140 and the U.S. and PCT applications cited therein and An et al., Monoclonal Antibodies (mAbs) 1: 6, 572-579 (2009); the entire disclosures of said patents are incorporated herein by reference.

[0127] "Hinge", "hinge domain" or "hinge region" or "antibody hinge region" refers to the domain of the heavy chain constant region connecting the CH1 domain and the CH2 domain, which includes the upper, middle and lower parts of the hinge (Roux et al., J. Immunol. 1998 161:4083). The hinge provides different levels of flexibility between the binding and effector regions of the antibody, and also provides a site for intermolecular disulfide bonds between the two heavy chain constant regions. For all IgG isotypes, the hinge used herein starts at Glu216 and ends at Gly237 (Roux et al., 1998, J. Immunol. 161:4083). The sequences of wild-type IgG1, IgG2, IgG3 and IgG4 hinges are known in the art. For example, see Kabat EA et al. (1991) Sequences of Proteins of Immunological Interest, 5th ed., U.S. Department of Health and Human Services, NIH Publication No. 91-3242; Vidarsson G. et al., Front Immunol. 5:520 (published online on October 20, 2014).

[0128] The term "CH1 domain" refers to the heavy chain constant region, which connects the variable domain to the hinge in the heavy chain constant domain. The CH1 domain used herein starts at A118 and ends at V215. The term "CH1 domain" includes wild-type CH1 domains, and naturally occurring variants thereof (e.g., allotypes). In the art, CH1 domain sequences of IgG1, IgG2, IgG3, and IgG4 (including wild-type and allotypes) are known. For example, see Kabat EA et al., (1991) supra, and Vidarsson G. et al. Frontiers in Immunology (Front Immunol.) 5: 520 (published online on October 20, 2014). Exemplary CH1 domains include CH1 domains with mutations that change the biological activity of antibodies (e.g., half-life), such as described in U.S. Patents (Publication No.: 20120100140) and U.S. Patents and publications cited therein and PCT publications.

[0129] The term "CH2 domain" refers to the heavy chain constant region connecting the hinge and the CH3 domain in the heavy chain constant domain. The CH2 domain used herein starts at P238 and ends at K340. The term "CH2 domain" includes wild-type CH2 domains, and naturally occurring variants thereof (e.g., allotypes). In the art, CH2 domain sequences of IgG1, IgG2, IgG3, and IgG4 (including wild-type and allotypes) are known. For example, see Kabat EA et al., (1991) supra, and Vidarsson G. et al. Frontiers in Immunology (Front Immunol.) 5: 520 (published online on October 20, 2014). Exemplary CH2 domains include CH2 domains with mutations that change the biological activity of antibodies (e.g., reduced half-life and / or Fc effector function), such as described in U.S. Patents (Publication No.: 20120100140) and U.S. Patents and publications cited therein and PCT publications.

[0130] The term "CH3 domain" refers to the heavy chain constant region, which is the C-terminus of the CH2 domain in the heavy chain constant domain. The CH3 domain used herein starts at G341 and ends at K447. The term "CH3 domain" includes wild-type CH3 domains, and naturally occurring variants thereof (e.g., allotypes). In the art, CH3 domain sequences of IgG1, IgG2, IgG3, and IgG4 (including wild-type and allotypes) are known. For example, see Kabat EA et al., (1991) supra, and Vidarsson G. et al. Frontiers in Immunology (Front Immunol.) 5: 520 (published online on October 20, 2014). Exemplary CH3 domains include CH3 domains with mutations that change the biological activity of antibodies (e.g., half-life), such as described in U.S. Patents (Publication No.: 20120100140) and U.S. Patents and publications cited therein and PCT publications.

[0131] As used herein, "isotype" refers to the antibody class encoded by the heavy chain constant region genes (e.g., IgG1, IgG2, IgG3, IgG4, IgM, IgAl, IgA2, IgD, and IgE antibodies).

[0132] "Allotype" refers to naturally occurring variants within a particular isotype group that differ in a few amino acids (e.g., see Jefferis et al. (2009) Monoclonal Antibodies (mAbs) 1:1). The antibodies described herein can be any allotype antibody. In the art, IgG1, IgG2, IgG3, and IgG4 allotypes are known. For example, see Kabat EA et al., (1991) supra; Vidarsson G. et al. Frontiers in Immunology (Front Immunol.) 5:520 (published online on October 20, 2014); and Lefranc MP, Monoclonal Antibodies (mAbs) 1:4, 1-7 (2009).

[0133] Herein, the phrases "an antibody that recognizes an antigen" and "an antibody specific for an antigen" are used interchangeably with the term "an antibody that specifically binds to an antigen."

[0134] As used herein, "isolated antibodies" refer to antibodies that are substantially free of other antibodies with different antigenic specificities (e.g., an isolated antibody that specifically binds to FAM19A5 is substantially free of antibodies that specifically bind to antigens other than FAM19A5). However, an isolated antibody that specifically binds to a FAM19A5 epitope may have cross-reactivity with other FAM19A5 proteins from different species.

[0135] "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 antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, "binding affinity" as used herein refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y can generally be expressed in terms of the dissociation constant (K D Affinity can be measured and / or expressed in a variety of ways known in the art, including but not limited to the equilibrium dissociation constant (K D ) and equilibrium association constant (K A ). D According to k 关 / k 开 The quotient of is calculated and expressed as molar concentration (M), while K A According to k 开 / k 关 The quotient of K is calculated. 开 refers to, for example, the binding rate constant of an antibody to an antigen, and k 关 Refers to, for example, the dissociation of antibodies and antigens. 开 and k 关It can be determined by techniques known to those of ordinary skill in the art, such as immunoassays (e.g., enzyme-linked immunosorbent assay (ELISA)), BIACORE TM Kinetic exclusion assay

[0136] As used herein, the terms "specific binding", "specific recognition", "specific binding", "selective binding", and "selective binding" are analogous terms in the context of antibodies and refer to molecules (e.g., antibodies) that bind to an antigen (e.g., an epitope or an immune complex), and such binding is understood by those skilled in the art. For example, by immunoassays, BIACORE TM , 3000 instrument (Sapidyne Instruments, Boise, ID) or other assays known in the art, it can be determined that a molecule that specifically binds to an antigen can bind to other peptides or polypeptides, generally with lower affinity. In a specific aspect, a molecule that specifically binds to an antigen has a K of 1. A At least 2 logs, 2.5 logs, 3 logs, 4 logs or greater than the K of the molecule when it binds to another antigen A .

[0137] Antibodies usually bind to their cognate antigens with high affinity and specificity, as shown by -5 Up to 10 -11 M or lower dissociation constant (K D ). It is generally believed that any value greater than about 10 -4 M of K D As used herein, an antibody that "specifically binds" an antigen refers to an antibody that binds the antigen with high affinity to substantially the same antigen, which means that when detected by, for example, immunoassays (e.g., ELISA) or surface plasmon resonance (SPR) techniques in BIACORE TM 2000 instrument with 10 -7 M or less, preferably 10 -8 M or less, even more preferably 10 -9 M or less, and most preferably 10 -8 M to 10 -10 M or lower K D , but does not bind unrelated antigens with high affinity.

[0138] As used herein, the term "antigen" refers to any natural or synthetic immunogenic substance, such as a protein, peptide or hapten. The antigen may be FAM19A5 or a fragment thereof.

[0139] As used herein, "epitope" is a term in the art, referring to a local region of an antigen to which an antibody can specifically bind. An epitope can be, for example, a continuous amino acid (linear or continuous epitope) of a polypeptide, or an epitope can be, for example, from two or more non-continuous regions (conformation, non-linearity, discontinuous or non-continuous epitopes) of one or more polypeptides. An epitope formed by continuous amino acids is usually, but not always, retained when exposed to a denaturing solvent, while an epitope formed by a tertiary folding is usually lost when treated with a denaturing solvent. An epitope generally includes at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 20 amino acids in a unique spatial conformation. It is well known in the art that methods for determining which epitopes are bound by a given antibody (i.e., epitope mapping) are used, and include, for example, immunoblotting and immunoprecipitation assays, wherein overlapping or continuous peptides from (e.g., from FAM19A5) are tested for reactivity with a given antibody (e.g., anti-FAM19A5 antibody). Methods for determining the spatial conformation of an epitope include techniques in the art and those described herein, for example, x-ray crystallography, two-dimensional nuclear magnetic resonance, and HDX-MS (see, for example, Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, ed. G.E. Morris (1996)).

[0140] In certain aspects, the epitope to which the antibody binds can be determined by, for example, NMR spectroscopy, X-ray diffraction crystallography studies, ELISA assays, hydrogen / deuterium exchange coupled to mass spectrometry (e.g., liquid chromatography electrospray mass spectrometry), array-based oligopeptide scanning assays, and / or mutagenesis mapping (e.g., site-directed mutagenesis mapping). For X-ray crystallography, crystallization can be accomplished by any method known in the art (e.g., Giege R et al., (1994) Acta Crystallogr D Biol Crystallogr, 50(Pt 4): 339-350; McPherson A (1990) Eur J Biochem, 189: 1-23; Chayen NE (1997) Structure, 5: 1269-1274; McPherson A (1976) J Biol Chem, 251: 6300-6303). Antibody:antigen crystals can be studied using well-known X-ray diffraction techniques and can be refined using computer software such as X-PLOR (Yale University, 1992, published by Molecular Simulations, Inc.; for example, see Meth Enzymol (1985) Nos. 114 and 115, edited by Wyckoff HW et al.; US2004 / 0014194), and BUSTER (Bricogne G (1993) Acta Crystallographica Sinica D: Biological Crystallography (Acta Crystallogr D Biol Crystallogr) 49 (Pt 1): 37-60; Bricogne G (1997) Meth Enzymol 276 N. 361-423, Carter CW; Roversi P et al., eds., (2000) Acta Crystallographica Sinica D: Biological Crystallography 56 (Pt 10): 1316-1323). Mutagenesis mapping studies can be performed using any method known to those skilled in the art. For mutagenesis techniques, including alanine scanning mutagenesis techniques, see, for example, Champe M et al., (1995) J Biol Chem 270:1388-1394, and Cunningham BC and Wells JA (1989) Science 244:1081-1085.

[0141] The term "epitope mapping" refers to the process of identifying molecular determinants for antibody-antigen recognition.

[0142] With reference to two or more antibodies, the term "binding to the same epitope" refers to the antibody binding to the same amino acid residue fragment determined by a given method. The technology used to determine whether an antibody binds to the "same epitope on FAM19A5" described herein includes, for example, epitope mapping methods, for example, x-ray analysis of crystals of antigen: antibody complexes, which provides atomic resolution and hydrogen / deuterium exchange mass spectrometry (HDX-MS) of epitopes. Other methods can monitor the binding of antibodies to antigen fragments or mutant variants of antigens, wherein it is generally believed that the binding loss caused by the modification of amino acid residues in the antigen sequence indicates the epitope component. In addition, epitope mapping can also be performed using computational combinatorial methods. These methods rely on the ability of related antibodies to affinity-free specific short peptides from a combined phage display peptide library. Antibodies with the same VH and VL or the same CDR1, 2 and 3 sequences are expected to bind to the same epitope.

[0143] An antibody that "competes with another antibody for binding to a target" refers to an antibody that (partially or completely) inhibits the binding of another antibody to a target. Using known competition experiments, it can be determined whether two antibodies compete with each other to bind to a target, i.e., whether one antibody inhibits the binding of another antibody to a target and the degree of inhibition. In certain aspects, an antibody competes with another antibody for target binding and inhibits the binding of another antibody to a target by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%. The level of inhibition or competition can be different, depending on which antibody is a "blocking antibody" (i.e., the first cold antibody incubated with the target). Competition assays are performed, for example, as described in Ed Harlow and David Lane, Cold Spring Harb Protoc; 2006; doi: 10.1101 / pdb.prot4277, or in Ed Harlow and David Lane, Using Antibodies (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA) (1999), Chapter 11. Competing antibodies bind to the same epitope, an overlapping epitope, or an adjacent epitope (e.g., as can be seen from steric hindrance).

[0144] Other competitive binding assays 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 labeled assays, solid phase direct labeled sandwich assays (see Harlow and Lane, Antibodies: A Laboratory Manual). Manual, Cold Spring Harbor Laboratory Press (1988); solid phase direct labeled RIA using 1-125 labeling (see Morel et al., Mol. Immunol. 25(1):7 (1988)); solid phase direct biotin-avidin EIA (Cheung et al., Virology 176:546 (1990)); and directly labeled RIA. (Moldenhauer et al., Scand. J. Immunol. 32:77 (1990)).

[0145] "Bispecific" or "bifunctional antibodies" are artificial hybrid antibodies with two different heavy chain / light chain pairs and two different binding sites. Bispecific antibodies can be produced by a variety of methods, including fusion of hybridomas or linking of Fab' fragments. For example, see Songsivilai and Lachmann, Clin. Exp. Immunol. 79: 315-321 (1990); Kostelny et al., J. Immunol. 148, 1547-1553 (1992).

[0146] The term "monoclonal antibody" as used herein refers to an antibody that exhibits a single binding specificity and affinity for a particular epitope, or an antibody composition in which all antibodies exhibit a single binding specificity and affinity for a particular epitope. Thus, the term "human monoclonal antibody" refers to an antibody or antibody composition that exhibits a single binding specificity and has variable and optional constant regions derived from human germline immunoglobulin sequences. In certain aspects, human monoclonal antibodies are produced by hybridomas that include B cells obtained from transgenic non-human animals, such as transgenic mice, whose genome includes human heavy chain transgenes and light chain transgenes fused to immortalized cells.

[0147] As used herein, the term "recombinant human antibody" includes all human antibodies prepared, expressed, generated or isolated by recombinant methods, such as (a) antibodies isolated from transgenic or transchromosomal animals (e.g., mice) for human immunoglobulin genes or hybridomas prepared therefrom, (b) antibodies isolated from host cells transformed to express antibodies (e.g., transfectomas), (c) antibodies isolated from recombinant, combinatorial human antibody libraries, and (d) antibodies prepared, expressed, generated or isolated by any other method involving splicing human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies include variable and constant regions that utilize specific human germline immunoglobulin sequences encoded by germline genes, but include subsequent rearrangements and mutations that occur, for example, during antibody maturation. It is known in the art (e.g., see Lonberg (2005) Nature Biotech. 23(9): 1117-1125) that variable regions contain antigen binding domains that are encoded by various genes that rearrange to form antibodies specific for foreign antigens. In addition to rearrangement, the variable region can be further modified by multiple single amino acid changes (called somatic mutations or hypermutations), which increase the affinity of the antibody for foreign antigens. The constant region will change in further response to the antigen (i.e., isotype switching). Therefore, in response to the antigen, the rearranged and somatically mutated nucleic acid molecules encoding the light and heavy chain immunoglobulin polypeptides cannot have sequence identity with the original nucleic acid molecules, but are substantially identical or similar (i.e., have at least 80% identity).

[0148] "HuMAb" refers to an antibody with a variable region, wherein the framework region and the CDR region are derived from human germline immunoglobulin sequences. In addition, if the antibody comprises a constant region, the constant region is also derived from human germline immunoglobulin sequences. The antibodies described herein may include amino acid residues that are not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutations in vivo). However, the term "human antibody" as used herein is not intended to include antibodies in which the CDR sequences derived from the germline of another mammalian species (such as a mouse) have been transplanted to human framework sequences. The term "human" antibody is synonymous with a "fully human" antibody.

[0149] A "humanized" antibody refers to an antibody in which some, most or all of the amino acids outside the CDR domain of a non-human antibody are replaced by the corresponding amino acids derived from human immunoglobulins. In some aspects of the humanized form of an antibody, some, most or all of the amino acids outside the CDR domain have been replaced with amino acids from human immunoglobulins, while some, most or all of the amino acids in one or more CDR regions have not changed. As long as the ability of the antibody to bind to a specific antigen is not eliminated, small additions, deletions, insertions, substitutions or modifications may be made to the amino acids. A "humanized" antibody retains an antigenic specificity similar to that of the original antibody.

[0150] A "chimeric antibody" refers to an antibody in which the variable region is derived from one species and the constant region is derived from another species, for example, an antibody in which the variable region is derived from a mouse antibody and the constant region is derived from a human antibody.

[0151] As used herein, the term "cross-reactivity" refers to the ability of the antibodies described herein to bind to FAM19A5 from different species. For example, an antibody described herein that binds to human FAM19A5 may also bind to FAM19A5 of another species (e.g., mouse FAM19A5). The cross-reactivity used herein can be measured by detecting specific reactivity with purified antigen in a binding assay (e.g., SPR, ELISA), or by binding to or otherwise functionally interacting with cells that physiologically express FAM19A5. Methods for determining cross-reactivity include standard binding assays described herein, for example, using BIACORE TM 2000SPR instrument (Biacore AB, Uppsala, Sweden), through BIACORE TM Surface plasmon resonance (SPR) analysis, or flow cytometry techniques.

[0152] The term "naturally occurring" as applied to an object herein means that an object can be found in nature. For example, a polypeptide or polynucleotide sequence present in an organism (including a virus) that can be isolated from a source in nature and has not been intentionally modified by man in the laboratory is naturally occurring.

[0153] "Polypeptide" refers to a chain of at least two consecutively linked amino acid residues, with no upper limit on the length of the chain. One or more amino acid residues in 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.

[0154] As used herein, the term "nucleic acid molecule" is intended to include DNA molecules and RNA molecules. The nucleic acid molecule may be a single-stranded or double-stranded nucleic acid molecule, and may also be a cDNA.

[0155] The term "vector" as used herein refers to a nucleic acid molecule capable of transporting another nucleic acid connected thereto. One type of vector is a "plasmid", which refers to a circular double-stranded DNA loop, wherein additional DNA fragments can be connected. Another type of vector is a viral vector, wherein additional DNA fragments can be connected to the viral genome. Some vectors can replicate autonomously in the host cells into which they are introduced (e.g., bacterial vectors and additional mammalian vectors with bacterial replication origins). Other vectors (e.g., non-additional mammalian vectors) can be integrated into the genome of the host cell after being introduced into the host cell, thereby replicating together with the host genome. In addition, some vectors can guide the expression of genes operably connected thereto. This vector is referred to as a "recombinant expression vector" (or simply "expression vector") in this article. Generally, expression vectors used for recombinant DNA technology are usually present in the form of plasmids. Since plasmids are the most commonly used vector forms, in this specification, "plasmids" and "vectors" can be used interchangeably. However, other forms of expression vectors are also included, such as viral vectors (e.g., replication-defective retroviruses, adenoviruses, and adeno-associated viruses), which can provide equivalent functions.

[0156] The term "recombinant host cell" (or simply "host cell") as used herein refers to a cell that includes 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 refer not only to the specific subject cell, but also to the progeny of such cells. Because certain modifications may occur in subsequent generations due to mutations or environmental influences, such progeny may not actually be identical to the parent cell, but are still included in the scope of the term "host cell" as used herein.

[0157] As used herein, the term "linked" refers to the association of two or more molecules. The link may be covalent or non-covalent. It may also be genetic (i.e., recombinant fusion) linked. Such a link may be achieved using a variety of techniques recognized in the art, such as chemical conjugation and recombinant protein production.

[0158] As used herein, the term "therapeutically effective amount" refers to an amount of a drug, alone or in combination with another therapeutic agent, that is effective to "treat" a disease or condition in an individual or to reduce the risk, likelihood, or occurrence of a disease or condition (e.g., such as diabetic retinopathy or age-related macular degeneration). A "therapeutically effective amount" includes an amount of a drug or therapeutic agent that provides some improvement or benefit to an individual suffering from or at risk of suffering from a disease or condition associated with fibrosis. Thus, a "therapeutically effective" amount is an amount that reduces the risk, likelihood, or occurrence of a disease or condition, or provides some relief, alleviation, and / or reduction in at least one indicator, and / or decreases at least one clinical symptom of a disease or condition.

[0159] II. Treatment of Retinopathy and / or Macular Degeneration

[0160] Disclosed herein are methods for treating eye conditions in individuals in need thereof. More specifically, the methods disclosed herein can be used to treat retinopathy, especially diabetic retinopathy. The methods disclosed herein can also be used to treat macular degeneration, especially age-related macular degeneration. As described below, eye diseases such as diabetic retinopathy and age-related macular degeneration are associated with reduced retinal potential. Therefore, also provided herein are methods for improving retinal potential in individuals in need thereof (e.g., with diabetic retinopathy or age-related macular degeneration).

[0161] In some aspects, the methods disclosed herein include administering to an individual an antagonist for a FAM19A5 protein ("FAM19A5 antagonist"). In some aspects, a FAM19A5 antagonist is an antisense oligonucleotide, siRNA, shRNA, miRNA, dsRNA, aptamer, PNA, or a vector comprising them that targets FAM19A5. In other aspects, a FAM19A5 antagonist is an antibody or an antigen-binding portion thereof ("anti-FAM19A5 antibody") that specifically binds to a FAM19A5 protein, a polynucleotide encoding an anti-FAM19A5 antibody, or a vector comprising a polynucleotide thereof. In some aspects, an anti-FAM19A5 antibody binds to a FAM19A5 protein and reduces FAM19A5 activity.

[0162] Individuals with retinopathy (e.g., diabetic retinopathy) and / or macular degeneration (e.g., age-related macular degeneration) may exhibit one or more of the following characteristics within the retina: (i) decreased retinal potential, (ii) pericyte loss, (iii) increased acellular capillary formation, (iv) increased vascular congestion (e.g., within the nerve fiber layer of the retina), (v) increased apoptotic cells (e.g., within the nerve fiber layer and / or inner nuclear layer of the retina), (vi) increased retinal and / or choroidal neovascularization, and (vii) increased astrocyte formation (e.g., increased PDGF staining within the nerve fiber layer and / or ganglion cell layer within the retina). See Examples 6-10.

[0163] Therefore, without being bound by any theory, by reducing, alleviating and / or reversing one or more of the above properties, the FAM19A5 antagonists disclosed herein (e.g., anti-FAM19A5 antibodies) can treat retinopathy and / or macular degeneration. In some aspects, the FAM19A5 antagonists can neutralize the FAM19A5 protein, such as a FAM19A5 neutralizer. On the other hand, the FAM19A5 antagonists are anti-FAM19A5 antibodies. In some aspects, the anti-FAM19A5 antibodies are neutralizing antibodies.

[0164] In some aspects, FAM19A5 antagonists can improve the retinal potential of an individual (e.g., suffering from diabetic retinopathy or age-related macular degeneration). In some aspects, compared to a reference value (e.g., a corresponding value of a DR or AMD individual not treated with a FAM19A5 antagonist disclosed herein), the improvement of the retinal potential includes an increase in A wave, B wave, and / or oscillatory potential values. In some aspects, compared to a reference value, A wave, B wave, and / or oscillatory potential values ​​increase by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

[0165] In some aspects, FAM19A5 antagonists disclosed herein (e.g., anti-FAM19A5 antibodies) can reduce or prevent pericyte loss in the retina of an individual (e.g., suffering from diabetic retinopathy or age-related macular degeneration). In some aspects, pericyte loss in the retina is at least reduced by about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% compared to a reference value (e.g., a corresponding value of a DR or AMD individual not treated with a FAM19A5 antagonist disclosed herein).

[0166] In some aspects, the FAM19A5 antagonists disclosed herein can reduce and / or inhibit the formation of acellular capillaries in the retina of an individual (e.g., suffering from diabetic retinopathy or age-related macular degeneration). In some aspects, compared to a reference value (e.g., a corresponding value of a DR or AMD individual without the treatment of a FAM19A5 antagonist disclosed herein), the formation of acellular capillaries is at least reduced and / or inhibited by about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%.

[0167] In some aspects, FAM19A5 antagonists (e.g., anti-FAM19A5 antibodies) can reduce and / or inhibit vascular congestion in the retina of an individual (e.g., suffering from diabetic retinopathy or age-related macular degeneration). In some aspects, vascular congestion in the retina is reduced and / or inhibited by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% compared to a reference value (e.g., a corresponding value of a DR or AMD individual not treated with a FAM19A5 antagonist disclosed herein).

[0168] In some aspects, the FAM19A5 antagonists disclosed herein can reduce and / or inhibit apoptosis of retinal cells in individuals (e.g., those suffering from diabetic retinopathy or age-related macular degeneration). In some aspects, the frequency of apoptosis of retinal cells is at least reduced by about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% compared to a reference value (e.g., a corresponding value of a DR or AMD individual not treated with a FAM19A5 antagonist disclosed herein).

[0169] In some aspects, FAM19A5 antagonists can reduce and / or inhibit neovascularization (e.g., retinal and / or choroidal neovascularization) in the retina of an individual (e.g., suffering from diabetic retinopathy or age-related macular degeneration). In some aspects, neovascularization (e.g., retinal and / or choroidal neovascularization) in the retina is reduced and / or inhibited by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% compared to a reference value (e.g., a corresponding value of a DR or AMD individual not treated with a FAM19A5 antagonist disclosed herein).

[0170] In some aspects, FAM19A5 antagonists are used to reduce and / or suppress PDGF expression in individual (e.g., suffering from diabetic retinopathy or age-related macular degeneration) retina. In some aspects, compared with a reference value (e.g., a corresponding value of a DR or AMD individual treated with FAM19A5 antagonists disclosed herein), PDGF expression in the retina is at least reduced and / or suppressed by about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%. In some aspects, reducing and / or suppressing PDGF expression is relevant to the reduction of astrocyte formation in the retina of an individual.

[0171] In certain aspects, the term "reference" as used herein refers to a corresponding individual (e.g., with diabetic retinopathy or age-related macular degeneration) who has not received a composition disclosed herein (e.g., a FAM19A5 antagonist, e.g., an anti-FAM19A5 antibody). The term "reference" may also refer to the same individual (e.g., with diabetic retinopathy or age-related macular degeneration), but before the composition disclosed herein is administered. In certain aspects, the term "reference" refers to the average number of individuals (e.g., with diabetic retinopathy or age-related macular degeneration).

[0172] In certain aspects, the retinopathy that can be treated by the present disclosure includes diabetic retinopathy. In certain aspects, the diabetic retinopathy is non-proliferative diabetic retinopathy (NPDR). In other aspects, the diabetic retinopathy is proliferative diabetic retinopathy (PDR). In certain aspects, the diabetic retinopathy is diabetic maculopathy. In more aspects, the diabetic retinopathy is diabetic macular edema. In certain aspects, the diabetic retinopathy is any retinopathy, associated with ischemic damage within the retina.

[0173] In some aspects, the macular degeneration that can be treated by the present disclosure includes age-related macular degeneration (AMD). In some aspects, age-related macular degeneration is early AMD. In other aspects, age-related macular degeneration is mid-term AMD. In more aspects, age-related macular degeneration is late AMD (i.e., geographic atrophy). In some aspects, age-related macular degeneration is dry (non-exudative) AMD. In other aspects, age-related macular degeneration is wet (neovascular or exudative) AMD.

[0174] In certain aspects, the individual that can be treated by the present method is a non-human animal, such as a rat or a mouse. In other aspects, the individual that can be treated by the methods disclosed herein is a human.

[0175] In some aspects, intraocular administration or any other application of compositions disclosed herein (e.g., FAM19A5 antagonists) can be delivered, to the fascia, conjunctiva, suprachoroidal space, vitreous and similar parts in the individual eye to deliver FAM19A5 antagonists disclosed herein (e.g., anti-FAM19A5 antibodies), bispecific molecules or immunoconjugates or their compositions. In some aspects, intraocular administration includes intravitreal administration.

[0176] When administering a FAM19A5 antagonist disclosed herein or a composition thereof (eg, an anti-FAM19A5 antibody), the dosage range is about 0.0001 to 100 mg / kg.

[0177] In certain aspects, a FAM19A5 antagonist or a composition thereof may be administered in combination with one or more other agents (e.g., therapeutic agents) to treat retinopathy (e.g., diabetic retinopathy) and / or macular degeneration (e.g., age-related macular degeneration), such as standard therapy, e.g., (i) corticosteroids, (ii) anti-angiogenesis inhibitors (e.g., anti-VEGF inhibitors), (iii) laser therapy (e.g., laser photocoagulation therapy), (iv) antioxidants. Other currently available treatment options that may be used in combination with a FAM19A5 antagonist or a composition thereof include, but are not limited to, surgery (e.g., vitrectomy) and control of modifiable risk factors (e.g., lowering blood sugar and cholesterol levels, stopping smoking).

[0178] III. FAM19A5 antagonists

[0179] In certain aspects, the FAM19A5 antagonists useful for the present disclosure are antisense oligonucleotides, siRNA, shRNA, miRNA, dsRNA, aptamers, PNA (peptide nucleic acid) that specifically target FAM19A5, or vectors comprising them. In other aspects, the FAM19A5 antagonists are antibodies or antigen-binding portions thereof that specifically bind to FAM19A5 protein ("anti-FAM19A5 antibodies"), polynucleotides encoding anti-FAM19A5 antibodies, or vectors comprising their polynucleotides.

[0180] In the methods disclosed herein, useful antibodies include monoclonal antibodies characterized by specific functional characteristics or properties. For example, the antibody specifically binds to human FAM19A5, including soluble FAM19A5 and membrane-bound FAM19A5. In addition to specifically binding to soluble and / or membrane-bound human FAM19A5, the antibodies described herein also (a) bind to FAM19A5 with a K of 10 nM or less. D Binds to soluble human FAM19A5; (b) with a K of 10 nM or lower D Binds to membrane-bound human FAM19A5; or both (a) and (b).

[0181] In certain aspects, an anti-FAM19A5 antibody or antigen-binding portion thereof specifically binds to soluble human FAM19A5 or membrane-bound human, e.g., K D For 10 -7 M or lower, 10 -8 M (10nM) or lower, 10 -9 M (1nM) or lower, 10 -10 M (0.1nM) or lower, 10 -11 M or lower, or 10 -12 M or lower, e.g. 10 -12 M-10 -7 M, 10 -11 M-10 -7 M, 10 -10 M-10 -7 M, or 10 -9 M-10 -7 M, for example 10 -12 M, 5X 10 -12 M, 10 -11 M, 5X 10 -11 M, 10 -10 M, 5X 10 -10 M, 10 -9 M, 5X10 -9M, 10 -8 M, 5X 10 -8 M, 10 -7 M, or 5X 10 -7 M. Standard assays known in the art can be used to evaluate the ability of antibodies of various species to bind to human FAM19A5, including, for example, ELISA, immunoblotting, and RIA. Suitable assays are described in detail in the Examples. The binding kinetics (e.g., binding affinity) of the antibodies can also be assessed by standard assays known in the art, such as by ELISA, BIACORE TM Analysis or

[0182] In certain aspects, the anti-FAM19A5 antibody, or antigen-binding portion thereof, is expressed as 10 -7 M or lower, 10 -8 M (10nM) or lower, 10 -9 M (1nM) or lower, 10 -10 M or lower, 10 -12 M-10 -7 M, 10 -11 M-10 -7 M, 10 -10 M-10 -7 M, 10 -9 M-10 -7 M or 10 -8 M-10 -7 M of K D Binds to soluble human FAM19A5. In certain aspects, the anti-FAM19A5 antibody or antigen-binding portion thereof binds to soluble human FAM19A5 with a K of 10 nM or less. D , such as 0.1 to 10 nM, 0.1 to 5 nM, 0.1 to 1 nM, 0.5 to 10 nM, 0.5 to 5 nM, 0.5 to 1 nM, 1 to 10 nM, 1 to 5 nM or 5 to 10 nM binding to soluble FAM19A5. In certain aspects, the anti-FAM19A5 antibody, or antigen-binding portion thereof, has a K of about 1 pM, 2 pM, 3 pM, 4 pM, 5 pM, 6 pM, 7 pM, 8 pM, 9 pM, 10 pM, 20 pM, 30 pM, 40 pM, 50 pM, 60 pM, 70 pM, 80 pM, 90 pM, 100 pM, 200 pM, 300 pM, 400 pM, 500 pM, 600 pM, 700 pM, 800 pM, or 900 pM, or about 1 nM, 2 nM, 3 nM, 4 nM, 5 nM, 6 nM, 7 nM, 8 nM, or 9 nM, or about 10 nM, 20 nM, 30 nM, 40 nM, 50 nM, 60 nM, 70 nM, 80 nM, or 90 nM as determined by ELISA. DSpecifically binds soluble human FAM19A5.

[0183] In certain aspects, the anti-FAM19A5 antibody, or antigen-binding portion thereof, is expressed as 10 -7 M or lower, 10 -8 M (10nM) or lower, 10 -9 M (1nM) or lower, 10 -10 M or lower, 10 -12 M-10 -7 M, 10 -11 M-1 -7 M, 10 -10 M-10 -7 M, 10 -9 M-10 -7 M or 10 -8 M-10 -7 M of K D In certain aspects, the anti-FAM19A5 antibody, or antigen-binding portion thereof, has a K of 10 nM or less as determined by ELISA. D , such as 0.1 to 10 nM, 0.1 to 5 nM, 0.1 to 1 nM, 0.5 to 10 nM, 0.5 to 5 nM, 0.5 to 1 nM, 1 to 10 nM, 1 to 5 nM or 5 to 10 nM specifically binds to membrane-bound human FAM19A5. In certain aspects, the anti-FAM19A5 antibody, or antigen-binding portion thereof, has a K of about 1 pM, 2 pM, 3 pM, 4 pM, 5 pM, 6 pM, 7 pM, 8 pM, 9 pM, 10 pM, 20 pM, 30 pM, 40 pM, 50 pM, 60 pM, 70 pM, 80 pM, 90 pM, 100 pM, 200 pM, 300 pM, 400 pM, 500 pM, 600 pM, 700 pM, 800 pM, or 900 pM, or about 1 nM, 2 nM, 3 nM, 4 nM, 5 nM, 6 nM, 7 nM, 8 nM, or 9 nM, or about 10 nM, 20 nM, 30 nM, 40 nM, 50 nM, 60 nM, 70 nM, 80 nM, or 90 nM as determined by ELISA. D Binding to membrane-bound human FAM19A5.

[0184] In certain aspects, an anti-FAM19A5 antibody or antigen binding portion thereof useful in the methods disclosed herein cross-competes for binding to (or inhibits binding of) a human FAM19A5 epitope with an anti-FAM19A5 antibody comprising CDRs or variable regions disclosed herein.

[0185] In certain aspects, the anti-FAM19A5 antibody or antigen-binding portion thereof inhibits binding of a reference antibody comprising a heavy chain CDR1, CDR2 and CDR3 and a light chain CDR1, CDR2 and CDR3, (i) wherein the heavy chain CDR1, CDR2 and CDR3 of the reference antibody comprise the amino acid sequences of SEQ ID NO: 11, SEQ ID NO: 12 and SEQ ID NO: 13, respectively, and the light chain CDR1, CDR2 and CDR3 of the reference antibody comprise the amino acid sequences of SEQ ID NO: 23, SEQ ID NO: 24 and SEQ ID NO: 25; (ii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 14, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 15, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 16, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 26, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 27, the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 28 NO:28; (iii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO:17, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO:18, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:19, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:29, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:30, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:31; (iv) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO:20, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO:21, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:22, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:32, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:33, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:34; (v) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO:89, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: ID NO:90, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:91, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:92, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:93, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:94;(vi) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 95, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 96, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 97, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 98, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 99, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 100; (vii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 101, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 102, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 103, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 104, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 105, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 106; (viii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 107, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 108 NO: 108, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 109, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 110, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 111, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 112; (ix) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 113, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 114, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 115, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 116, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 117, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 118; (x) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 119, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 120, and the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 121 NO: 121, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 122, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 123, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 124;(xi) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 125, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 126, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 127, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 128, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 129, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 130; (xii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 131, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 132, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 133, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 134, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 135, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 136; (xiii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 137, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 138 NO: 138, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 139, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 140, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 141, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 142; (xiv) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 143, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 144, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 145, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 146, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 147, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 148; or (xv) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 149, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 150, and the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 151. NO: 151, the light chain CDR1 includes the amino acid sequence of SEQ ID NO: 152, the light chain CDR2 includes the amino acid sequence of SEQ ID NO: 153, and the light chain CDR3 includes the amino acid sequence of SEQ ID NO: 154. ;

[0186] In certain aspects, the reference antibody comprises (a) heavy and light chain variable region sequences comprising SEQ ID NOs: 35 and 39, respectively; (b) heavy and light chain variable region sequences comprising SEQ ID NOs: 36 and 40, respectively; (c) heavy and light chain variable region sequences comprising SEQ ID NOs: 37 and 41, respectively; (d) heavy and light chain variable region sequences comprising SEQ ID NOs: 38 and 42, respectively; (e) heavy and light chain variable region sequences comprising SEQ ID NOs: 155 and 166, respectively; (f) heavy and light chain variable region sequences comprising SEQ ID NOs: 156 and 167, respectively; (g) heavy and light chain variable region sequences comprising SEQ ID NOs: 157 and 168, respectively; (h) heavy and light chain variable region sequences comprising SEQ ID NOs: 158 and 169, respectively; (i) heavy and light chain variable region sequences comprising SEQ ID NOs: 159 and 170, respectively; (j) heavy and light chain variable region sequences comprising SEQ ID NOs: 171 and 172, respectively. NO: 160 and 171; (k) heavy chain and light chain variable region sequences comprising SEQ ID NO: 161 and 172, respectively; (l) heavy chain and light chain variable region sequences comprising SEQ ID NO: 162 and 173, respectively; (m) heavy chain and light chain variable region sequences comprising SEQ ID NO: 163 and 174, respectively; (n) heavy chain and light chain variable region sequences comprising SEQ ID NO: 164 and 175, respectively; or (o) heavy chain and light chain variable region sequences comprising SEQ ID NO: 165 and 176, respectively.

[0187] In certain aspects, the anti-FAM19A5 antibody or its antigen-binding portion inhibits the binding of such reference antibody to human FAM19A5 by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90%, or 100%. Competitive antibodies bind to the same epitope, overlapping epitope or adjacent epitope (e.g., demonstrated by steric hindrance). Competition experiments known in the art, such as RIA and EIA, can be used to determine whether two antibodies compete with each other for binding to the target.

[0188] In certain aspects, the anti-FAM19A5 antibody, or antigen-binding portion thereof, binds to the same FAM19A5 epitope as a reference antibody disclosed herein comprising a heavy chain CDR1, CDR2 and CDR3 and a light chain CDR1, CDR2 and CDR3, (i) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 11, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 12, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 13, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 23, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 24, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 25; (ii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 14, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 15, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 16, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 26, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: NO:27, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:28; (iii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO:17, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO:18, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:19, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:29, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:30, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:31; (iv) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO:20, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO:21, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:22, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:32, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:33, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:34; (v) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: NO:89, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO:90, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:91, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO:92, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO:93, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO:94;(vi) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 95, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 96, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 97, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 98, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 99, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 100; (vii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 101, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 102, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 103, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 104, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 105, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 106; (viii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 107, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 108 NO: 108, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 109, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 110, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 111, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 112; (ix) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 113, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 114, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 115, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 116, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 117, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 118; (x) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 119, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 120, and the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 121 NO: 121, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 122, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 123, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 124;(xi) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 125, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 126, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 127, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 128, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 129, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 130; (xii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 131, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 132, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 133, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 134, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 135, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 136; (xiii) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 137, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 138 NO: 138, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 139, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 140, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 141, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 142; (xiv) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 143, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 144, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 145, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 146, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 147, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 148; or (xv) wherein the heavy chain CDR1 comprises the amino acid sequence of SEQ ID NO: 149, the heavy chain CDR2 comprises the amino acid sequence of SEQ ID NO: 150, and the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO: 151 NO: 151, the light chain CDR1 includes the amino acid sequence of SEQ ID NO: 152, the light chain CDR2 includes the amino acid sequence of SEQ ID NO: 153, and the light chain CDR3 includes the amino acid sequence of SEQ ID NO: 154. ;

[0189] In certain aspects, the reference antibody comprises (a) heavy and light chain variable region sequences comprising SEQ ID NOs: 35 and 39, respectively; (b) heavy and light chain variable region sequences comprising SEQ ID NOs: 36 and 40, respectively; (c) heavy and light chain variable region sequences comprising SEQ ID NOs: 37 and 41, respectively; (d) heavy and light chain variable region sequences comprising SEQ ID NOs: 38 and 42, respectively; (e) heavy and light chain variable region sequences comprising SEQ ID NOs: 155 and 166, respectively; (f) heavy and light chain variable region sequences comprising SEQ ID NOs: 156 and 167, respectively; (g) heavy and light chain variable region sequences comprising SEQ ID NOs: 157 and 168, respectively; (h) heavy and light chain variable region sequences comprising SEQ ID NOs: 158 and 169, respectively; (i) heavy and light chain variable region sequences comprising SEQ ID NOs: 159 and 170, respectively; (j) heavy and light chain variable region sequences comprising SEQ ID NOs: 171 and 172, respectively. NO: 160 and 171; (k) heavy chain and light chain variable region sequences comprising SEQ ID NO: 161 and 172, respectively; (l) heavy chain and light chain variable region sequences comprising SEQ ID NO: 162 and 173, respectively; (m) heavy chain and light chain variable region sequences comprising SEQ ID NO: 163 and 174, respectively; (n) heavy chain and light chain variable region sequences comprising SEQ ID NO: 164 and 175, respectively; or (o) heavy chain and light chain variable region sequences comprising SEQ ID NO: 165 and 176, respectively.

[0190] For example, techniques for determining whether two antibodies bind to the same epitope include epitope mapping methods, such as crystallographic x-ray analysis of antigen:antibody complexes, which provides atomic resolution of the epitope and hydrogen / deuterium exchange mass spectrometry (HDX-MS), methods for monitoring antibody binding to antigen fragments or mutant variants of the antigen (in which loss of binding resulting from modification of amino acid residues within the antigen sequence is generally considered to be an indication of epitope composition), and computational combined methods of epitope mapping.

[0191] For example, an anti-FAM19A5 antibody or antigen binding portion thereof useful in the methods disclosed herein can bind to at least one epitope of mature human FAM19A5 by binding the antibody to a fragment of human FAM19A5. In certain aspects, an anti-FAM19A5 antibody or antigen binding portion thereof binds to a fragment within the amino acid sequence of TLDRDSSQPRRTIARQTARC (SEQ ID NO: 6 or amino acid residues 42 to 61 of SEQ ID NO: 2), such as an epitope having at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids of SEQ ID NO: 6. In certain aspects, an anti-FAM19A5 antibody or an antigen binding portion thereof binds to SEQ ID NO: 2 at one or more amino acids corresponding to amino acid residues 46 to 51 (i.e., DSSQPR), such as amino acid residues 46, 50 and 52 (i.e., D---PR), such as amino acid residues 46, 47, 48 and 50 (i.e., DSS-P). In certain aspects, an anti-FAM19A5 antibody or an antigen binding portion thereof binds to a fragment within the amino acid sequence of CDMLPCLEGEGCDLLINRSG (SEQ ID NO: 9 or amino acids 90 to 109 of SEQ ID NO: 2), such as an epitope having at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids of SEQ ID NO: 9. In certain aspects, an anti-FAM19A5 antibody, or antigen-binding portion thereof, binds to SEQ ID NO:9 at one or more amino acid residues 99 to 107 (i.e., EGCDLLINR), e.g., amino acid residues 102, 103, 105, and 107 (i.e., DL-IR), e.g., amino acid residues 99, 100, 102, 103, 105, and 107 (i.e., EG-DL-IR), e.g., amino acid residues 99, 100, and 107 (i.e., EG------R) of SEQ ID NO:4.

[0192] In certain aspects, the at least one epitope has an amino acid sequence that is at least 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 6. In certain aspects, the at least one epitope has an amino acid sequence that is at least 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to SEQ ID NO: 9.

[0193] In certain aspects, the anti-FAM19A5 antibody, or an antigen binding portion thereof, binds only to the human FAM19A5 epitope of SEQ ID NO: 5, 6, 7, 8, 9 or 10, or a fragment within the amino acid sequence of SEQ ID NO: 5, 6, 7, 8, 9 or 10, such as an epitope having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids of SEQ ID NO: 5, 6, 7, 8, 9 or 10.

[0194] In certain aspects, the anti-FAM19A5 antibodies or antigen-binding portions thereof of the present disclosure bind to SEQ ID NO: 6 or a fragment thereof in its native conformation (i.e., not denatured). In certain aspects, the anti-FAM19A5 antibodies or antigen-binding portions thereof of the present disclosure bind to SEQ ID NO: 9 or a fragment thereof in its native conformation (i.e., not denatured). In other aspects, the at least one epitope has an amino acid sequence that is at least 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 5, and has an amino acid sequence that is at least 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or about 100% identical to SEQ ID NO: 10. In certain aspects, the anti-FAM19A5 antibodies or antigen-binding portions thereof simultaneously bind to glycosylated and unglycosylated human FAM19A5.

[0195] In certain aspects, the anti-FAM19A5 antibody or antigen binding portion thereof further binds to one or more additional FAM19A5 epitopes. Thus, certain anti-FAM19A5 antibodies or antigen binding portions thereof bind to the epitope of SEQ ID NO: 6 and an additional epitope or the epitope of SEQ ID NO: 9 and an additional epitope. Other anti-FAM19A5 antibodies or antigen binding portions thereof may bind to the epitope of SEQ ID NO: 5, SEQ ID NO: 9, and an additional epitope. In certain aspects, the anti-FAM19A5 antibody or antigen binding portion thereof binds to the epitope of SEQ ID NO: 6, the epitope of SEQ ID NO: 10, and an additional epitope. In certain aspects, the one or more additional FAM19A5 epitopes are selected from QLAAGTCEIVTLDR (SEQ ID NO: 5, epitope F1), TLDRDSSQPRRTIARQTARC (SEQ ID NO: 6, epitope F2), TARCACRKGQIAGTTRARPA (SEQ ID NO: 7, epitope F3), ARPACVDARIIKTKQWCDML (SEQ ID NO: 8, epitope F4), CDMLPCLEGEGCDLLINRSG (SEQ ID NO: 9, epitope F5) or NRSGWTCTQPGGRIKTTTVS (SEQ ID NO: 10, epitope F6), or a fragment located within the amino acid sequence of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9 or SEQ ID NO: 10, or any combination thereof. Fragments located within the amino acid sequence of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10, including fragments having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids of any one of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10. In some aspects, the one or more additional FAM19A5 epitopes are selected from SEQ ID NO: 5, 6, 7, 8, 9 or 10, or a fragment located within the amino acid sequence of SEQ ID NO: 5, 6, 7, 8, 9 or 10, for example, a fragment having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids of SEQ ID NO: 5, 6, 7, 8, 9 or 10 or any combination thereof.In some aspects, anti-FAM19A5 antibodies or antigen binding portions thereof of the present disclosure bind to any one of one or more additional epitopes located in its native conformation (i.e., not denatured). In some aspects, anti-FAM19A5 antibodies or antigen binding portions thereof simultaneously bind to one or more additional FAM19A5 epitopes of glycosylation and non-glycosylation.

[0196] In certain aspects, an anti-FAM19A5 antibody, or an antigen binding portion thereof, binds to at least one FAM19A5 epitope identified as EP2, EP4, and / or EP8, wherein EP2 comprises, consists essentially of, or consists of the amino acids DSSQP (SEQ ID NO: 66), wherein EP4 comprises, consists essentially of, or consists of the amino acids ARCACRK (SEQ ID NO: 68), wherein EP8 comprises, consists essentially of, or consists of the amino acids TCTQPGGR (SEQ ID NO: 72). In certain aspects, the at least one epitope has an amino acid sequence that is at least 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to EP2, EP4, or EP8. In certain aspects, an anti-FAM19A5 antibody, or an antigen binding portion thereof, binds only to EP2. In certain aspects, an anti-FAM19A5 antibody, or an antigen binding portion thereof, binds to EP4 and EP8.

[0197] In certain aspects, an anti-FAM19A5 antibody or an antigen binding portion thereof binds to at least one FAM19A5 epitope identified as EP6, EP7 or EP8, wherein EP6 comprises the amino acids KTKQWCDML (SEQ ID NO: 70), wherein EP7 comprises the amino acids GCDLLINR (SEQ ID NO: 71), wherein EP8 comprises the amino acids TCTQPGGR (SEQ ID NO: 72). In certain aspects, the at least one epitope has an amino acid sequence that is at least 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to EP6, EP7, or EP8. In certain aspects, an anti-FAM19A5 antibody or an antigen binding portion thereof binds only to EP6, EP7, or EP8. In certain aspects, an anti-FAM19A5 antibody or an antigen binding portion thereof binds to EP6, EP7, and EP8. In certain aspects, an anti-FAM19A5 antibody or an antigen binding portion thereof binds to EP7 and EP8. In certain aspects, the anti-FAM19A5 antibody, or antigen binding portion thereof, binds EP7.

[0198] In certain aspects, the anti-FAM19A5 antibody, or antigen binding portion thereof, binds one or more FAM19A5 epitopes selected from the group consisting of SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, and any combination thereof.

[0199] In certain aspects, provided herein is an antibody or its antigen-binding portion thereof that is combined with FAM19A5 (such as human FAM19A5) with an affinity 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or higher than that of another protein in FAM19A family, and the affinity is measured by, for example, immunoassay (such as ELISA), surface plasmon resonance or kinetic exclusion assay. In a specific aspect, provided herein is an antibody or its antigen-binding portion thereof that is combined with FAM19A5 (such as human FAM19A5) and has no cross-reactivity with another protein in FAM19A family (such as immunoassay).

[0200] In certain aspects, the anti-FAM19A5 antibody is not a natural antibody, or is not a naturally occurring antibody. For example, the anti-FAM19A5 antibody has a different post-translational modification than a naturally occurring antibody (e.g., by having more, fewer, or different types of post-translational modifications).

[0201] IV. Exemplary Anti-FAM19A5 Antibodies

[0202] Specific antibodies that can be used in the methods disclosed herein are antibodies, such as monoclonal antibodies, having CDR and / or variable region sequences disclosed herein, and antibodies having at least 80% identity (e.g., at least 85%, at least 90%, at least 95% or at least 99% identity) to their variable region or CDR sequences. Tables 2 and 3 provide the amino acid sequences of the VH and VL CDRs of different anti-FAM19A5 antibodies, respectively. Using the Kabat numbering scheme (see above) 1-65, 3-2, 2-13, 1-28, P2-C12, 13B4, 13F7, 15A9, P1-A03, P1-A08, P1-F02, P2-A01, P2-A03, P2-F07, P2-F11, SS01-13, SS01-13-s5 and S5-2.GKNG, the CDRs of the following antibodies were identified. The CDRs of the following antibodies were identified using the IMGT numbering system (see above): 1-7A-IT, PI low, 1-30, 1-17, 1-32, 4-11, 6-10, 2-13D, 2-13D-37, 2-13D-37-1.5W-41, and 2-13D-37-3W-16. Tables 4 and 5 provide the VH and VL amino acid sequences of different anti-FAM19A5 antibodies of the present disclosure, respectively.

[0203] Table 2 Variable heavy chain CDR amino acid sequence (according to Kabat system)

[0204]

[0205]

[0206] Table 3 Variable light chain CDR amino acid sequence (according to Kabat system)

[0207]

[0208]

[0209] Table 4: Variable heavy chain amino acid sequences

[0210]

[0211]

[0212]

[0213] Table 5: Variable light chain amino acid sequences

[0214]

[0215]

[0216] Therefore, the present invention provides an isolated anti-FAM19A5 antibody or an antigen-binding portion thereof, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence of SEQ ID NO: 35-38, 155-165 or 232-240 (see Table 4). In other aspects, the isolated anti-FAM19A5 antibody or an antigen-binding portion thereof comprises a CDR of a heavy chain variable region selected from the group consisting of SEQ ID NO: 35-38, 155-165 or 232-240 (see Table 4).

[0217] Also provided herein is an isolated anti-FAM19A5 antibody or antigen binding portion thereof, comprising a heavy chain variable region and a light chain variable region, wherein the light chain variable region comprises an amino acid sequence of SEQ ID NO: 39-42, 166-176 or 241-250 (see Table 5). In other aspects, the isolated anti-FAM19A5 antibody or antigen binding portion thereof comprises a CDR of a light chain variable region selected from the group consisting of SEQ ID NO: 39-42, 166-176 or 241-250.

[0218] In certain aspects, the isolated anti-FAM19A5 antibody, or antigen binding portion thereof, comprises a CDR of a heavy chain variable region selected from the group consisting of SEQ ID NOs: 35-38, 155-165, or 232-240, and a CDR of a light chain variable region selected from the group consisting of SEQ ID NOs: 39-42, 166-176, or 241-250.

[0219] Also provided herein are isolated anti-FAM19A5 antibodies or antigen-binding portions thereof, comprising a heavy chain variable region and a light chain variable region, (i) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 35, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 39; (ii) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 36, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 40; (iii) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 37, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 41; (iv) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 38, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 42; (v) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 155, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 166; (vi) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: NO:156, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:167; (vii) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:157, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:168; (viii) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:158, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:169; (ix) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:159, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:170; (x) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:160, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:171; (xi) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:161, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:172; (xii) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:1 NO: 162, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 173; (xiii) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 163, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 174; (xiv) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 164, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 175;and (xv) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 165, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 176. In certain aspects, the isolated anti-FAM19A5 antibody or antigen-binding portion thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein VH comprises the VH sequence listed in Table 4, and VL comprises the VL sequence listed in Table 5. ;

[0220] Provided herein are isolated anti-FAM19A5 antibodies or antigen-binding portions thereof, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence that is 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%, at least about 99% or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 35-38, 155-165 or 232-240 (see Table 4).

[0221] Also provided herein is an isolated anti-FAM19A5 antibody or antigen-binding portion thereof, comprising a heavy chain variable region and a light chain variable region, wherein the light chain variable region comprises an amino acid sequence that is 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%, at least about 99% or about 100% identical to the amino acid sequence listed in SEQ ID NO: 39-42, 166-176 or 232-240 (see Table 5).

[0222] Also provided is an isolated anti-FAM19A5 antibody or an antigen-binding portion thereof, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence that is 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%, at least about 99% or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 35-38, 155-165 or 232-240 (see Table 4), wherein the light chain variable region comprises an amino acid sequence that is 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%, at least about 99% or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 39-42, 166-176 or 241-250 (see Table 5).

[0223] In certain aspects, the present disclosure provides an isolated anti-FAM19A5 antibody, or antigen-binding portion thereof, comprising:

[0224] (a) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 35 and 39, respectively;

[0225] (b) heavy and light chain variable region sequences comprising SEQ ID NOs: 36 and 40, respectively;

[0226] (c) heavy and light chain variable region sequences comprising SEQ ID NOs: 37 and 41, respectively;

[0227] (d) heavy and light chain variable region sequences comprising SEQ ID NOs: 38 and 42, respectively;

[0228] (e) heavy and light chain variable region sequences comprising SEQ ID NOs: 155 and 166, respectively;

[0229] (f) heavy and light chain variable region sequences comprising SEQ ID NOs: 156 and 167, respectively;

[0230] (g) heavy and light chain variable region sequences comprising SEQ ID NOs: 157 and 168, respectively;

[0231] (h) heavy and light chain variable region sequences comprising SEQ ID NOs: 158 and 169, respectively;

[0232] (i) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 159 and 170, respectively;

[0233] (j) heavy and light chain variable region sequences comprising SEQ ID NOs: 160 and 171, respectively;

[0234] (k) heavy and light chain variable region sequences comprising SEQ ID NOs: 161 and 172, respectively;

[0235] (l)) comprising the heavy and light chain variable region sequences of SEQ ID NOs: 162 and 173, respectively;

[0236] (m) heavy and light chain variable region sequences comprising SEQ ID NOs: 163 and 174, respectively;

[0237] (n) heavy and light chain variable region sequences comprising SEQ ID NOs: 164 and 175, respectively;

[0238] (o) heavy and light chain variable region sequences comprising SEQ ID NOs: 165 and 176, respectively;

[0239] (p) heavy and light chain variable region sequences comprising SEQ ID NOs: 232 and 241, respectively;

[0240] (q) heavy and light chain variable region sequences comprising SEQ ID NOs: 233 and 242, respectively;

[0241] (r) heavy and light chain variable region sequences comprising SEQ ID NOs: 234 and 242, respectively;

[0242] (s) heavy and light chain variable region sequences comprising SEQ ID NOs: 235 and 243, respectively;

[0243] (t) heavy and light chain variable region sequences comprising SEQ ID NOs: 236 and 244, respectively;

[0244] (u) heavy and light chain variable region sequences comprising SEQ ID NOs: 236 and 245, respectively;

[0245] (v) heavy and light chain variable region sequences comprising SEQ ID NOs: 236 and 246, respectively;

[0246] (w) heavy and light chain variable region sequences comprising SEQ ID NOs: 236 and 247, respectively;

[0247] (x) heavy and light chain variable region sequences comprising SEQ ID NOs: 236 and 248, respectively;

[0248] (y) heavy and light chain variable region sequences comprising SEQ ID NOs: 236 and 249, respectively;

[0249] (z) heavy and light chain variable region sequences comprising SEQ ID NOs: 237 and 250, respectively;

[0250] (aa) heavy and light chain variable region sequences comprising SEQ ID NOs: 238 and 250, respectively;

[0251] (bb) heavy and light chain variable region sequences comprising SEQ ID NOs: 239 and 250, respectively; or

[0252] (cc) heavy and light chain variable region sequences comprising SEQ ID NOs: 240 and 250, respectively.

[0253] In certain aspects, the anti-FAM19A5 antibodies or antigen-binding portions thereof of the present disclosure include (i) heavy chain CDR1s, CDR2s and CDR3s of 2-13 or combinations thereof, and / or light chain CDR1s, CDR2s and CDR3s of 2-13 or any combination thereof; (ii) heavy chain CDR1s, CDR2s and CDR3s of 3-2 or combinations thereof, and / or light chain CDR1s, CDR2s and CDR3s of 3-2 or any combination thereof; (iii) heavy chain CDR1s, CDR2s and CDR3s of 1-65 or combinations thereof, and / or light chain CDR1s, CDR2s and CDR3s of 1-65 or any combination thereof; (iv) heavy chain CDR1s, CDR2s and CDR3s of 1-28 or combinations thereof, and / or 1 -28's light chain CDR1, CDR2 and CDR3 or any combination thereof; (v) P2-C12's heavy chain CDR1, CDR2 and CDR3 or any combination thereof, and / or P2-C12's light chain CDR1, CDR2 and CDR3 or any combination thereof; (vi) 13B4's heavy chain CDR1, CDR2 and CDR3 or any combination thereof, and / or 13B4's light chain CDR1, CDR2 and CDR3 or any combination thereof; (vii) 13F7's heavy chain CDR1, CDR2 and CDR3 or any combination thereof, and / or 13F7's light chain CDR1, CDR2 and CDR3 or any combination thereof; (viii) 15A9's heavy chain CDR1, CDR2 and CDR3 or any combination thereof , and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of 15A9; (ix) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of P1-A03, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of P1-A03; (x) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of P1-A08, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of P1-A08; (xi) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of P1-F02, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of P1-F02; (xii) the heavy chain CDR1, CDR2 and CDR3 of P2-A01, CDR2 and CDR3 or any combination thereof, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of P2-A01; (xiii) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of P2-A03, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of P2-A03; (xiv) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of P2-F07, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of P2-F07; (xv) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of P2-F11, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of P2-F11;(xvi) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of SS01-13, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of SS01-13; (xvii) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of SS01-13-s5, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of SS01-13-s5; (xviii) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of S5-2.GKNG, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of S5-2.GKNG; (xix) The heavy chain CDR1, CDR2 and CDR3 or any combination thereof of 1-7A-IT, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of 1-7A-IT; (xx) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof with a low PI value, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof with a low PI value; (xxi) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of 1-30, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of 1-30; (xxii) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of 1-17, and / or or light chain CDR1s, CDR2s and CDR3s of 1-17 or any combination thereof; (xxiii) heavy chain CDR1s, CDR2s and CDR3s of 1-32 or any combination thereof, and / or light chain CDR1s, CDR2s and CDR3s of 1-32 or any combination thereof; (xxiv) heavy chain CDR1s, CDR2s and CDR3s of 4-11 or any combination thereof, and / or light chain CDR1s, CDR2s and CDR3s of 4-11 or any combination thereof; (xxv) heavy chain CDR1s, CDR2s and CDR3s of 6-10 or any combination thereof, and / or light chain CDR1s, CDR2s and CDR3s of 6-10 or any combination thereof; (xxviii) heavy chain CDR1s, CDR2s and CDR3s of 1-32 or any combination thereof, and / or light chain CDR1s, CDR2s and CDR3s of 1-32 or any combination thereof; (xxiv) heavy chain CDR1s, CDR2s and CDR3s of 4-11 or any combination thereof, and / or light chain CDR1s, CDR2s and CDR3s of 4-11 or any combination thereof; (xxviii) heavy chain CDR1s, CDR2s and CDR3s of 6-10 or any combination thereof, and / or light chain CDR1s, CDR2s and CDR3s of 6-10 or any combination thereof; i) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of 2-13D, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of 2-13D; (xxvii) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of 2-13D-37, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of 2-13D-37; (xxviii) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of 2-13D-37-1.5W-41, and / or the light chain CDR1, CDR2 and CDR3 or any combination thereof of 2-13D-37-1.5W-41;or (xxiv) the heavy chain CDR1, CDR2 and CDR3 of 2-13D-37-3W-16 or any combination thereof, and / or the light chain CDR1, CDR2 and CDR3 of 2-13D-37-3W-16 or any combination thereof. The amino acid sequences of VH CDR1, CDR2 and CDR3 of different anti-FAM19A5 antibodies disclosed herein are provided in Table 2. The amino acid sequences of VLCDR1, CDR2 and CDR3 of different anti-FAM19A5 antibodies disclosed herein are provided in Table 3. ;

[0254] In certain aspects, an anti-FAM19A5 antibody of the present disclosure, or an antigen-binding portion thereof, that specifically binds to human FAM19A5 comprises:

[0255] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 11; and / or

[0256] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 12; and / or

[0257] (c) VH CDR3 comprising the amino acid sequence of SEQ ID NO:13.

[0258] In specific aspects, the antibody, or antigen-binding portion thereof, comprises one, two or all three of the above-mentioned VHCDRs.

[0259] In certain aspects, an anti-FAM19A5 antibody or antigen-binding portion thereof that specifically binds to human FAM19A5 comprises:

[0260] (a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 23; and / or

[0261] (b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 24; and / or

[0262] (c) VL CDR3 comprising the amino acid sequence of SEQ ID NO:25.

[0263] In specific aspects, the antibody, or antigen-binding portion thereof, comprises one, two, or all three of the above-described VL CDRs.

[0264] In certain aspects, an anti-FAM19A5 antibody or antigen-binding portion thereof that specifically binds to human FAM19A5 comprises:

[0265] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 11;

[0266] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 12;

[0267] (c) a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 13;

[0268] (d) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 23;

[0269] (e) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 24; and / or

[0270] (f) VL CDR3 comprising the amino acid sequence of SEQ ID NO:25.

[0271] In certain aspects, an anti-FAM19A5 antibody of the present disclosure, or an antigen-binding portion thereof, that specifically binds to human FAM19A5 comprises:

[0272] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 14; and / or

[0273] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 15; and / or

[0274] (c) VH CDR3 comprising the amino acid sequence of SEQ ID NO:16.

[0275] In specific aspects, the antibody, or antigen-binding portion thereof, comprises one, two, or all three of the above-described VH CDRs.

[0276] In certain aspects, an anti-FAM19A5 antibody or antigen-binding portion thereof that specifically binds to human FAM19A5 comprises:

[0277] (a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 26; and / or

[0278] (b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 27; and / or

[0279] (c) VL CDR3 comprising the amino acid sequence of SEQ ID NO:28.

[0280] In specific aspects, the antibody, or antigen-binding portion thereof, comprises one, two, or all three of the above-described VL CDRs.

[0281] In certain aspects, an anti-FAM19A5 antibody or antigen-binding portion thereof that specifically binds to human FAM19A5 comprises:

[0282] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 14;

[0283] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 15;

[0284] (c) a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 16;

[0285] (d) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 26;

[0286] (e) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 27; and / or

[0287] (f) VL CDR3 comprising the amino acid sequence of SEQ ID NO:28.

[0288] In certain aspects, an anti-FAM19A5 antibody of the present disclosure, or an antigen-binding portion thereof, that specifically binds to human FAM19A5 comprises:

[0289] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 17; and / or

[0290] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 18; and / or

[0291] (c) VH CDR3 comprising the amino acid sequence of SEQ ID NO:19.

[0292] In specific aspects, the antibody, or antigen-binding portion thereof, comprises one, two, or all three of the above-described VH CDRs.

[0293] In certain aspects, an anti-FAM19A5 antibody or antigen-binding portion thereof that specifically binds to human FAM19A5 comprises:

[0294] (a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 29; and / or

[0295] (b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 30; and / or

[0296] (c) VL CDR3 comprising the amino acid sequence of SEQ ID NO:31.

[0297] In specific aspects, the antibody, or antigen-binding portion thereof, comprises one, two, or all three of the above-described VL CDRs.

[0298] In certain aspects, an anti-FAM19A5 antibody or antigen-binding portion thereof that specifically binds to human FAM19A5 comprises:

[0299] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 17;

[0300] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 18;

[0301] (c) a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 19;

[0302] (d) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 29;

[0303] (e) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 30; and / or

[0304] (f) VL CDR3 comprising the amino acid sequence of SEQ ID NO:31.

[0305] In certain aspects, an anti-FAM19A5 antibody of the present disclosure, or an antigen-binding portion thereof, that specifically binds to human FAM19A5 comprises:

[0306] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 20; and / or

[0307] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 21; and / or

[0308] (c) VH CDR3 comprising the amino acid sequence of SEQ ID NO:22.

[0309] In specific aspects, the antibody, or antigen-binding portion thereof, comprises one, two, or all three of the above-described VH CDRs.

[0310] In certain aspects, an anti-FAM19A5 antibody or antigen-binding portion thereof that specifically binds to human FAM19A5 comprises:

[0311] (a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 32; and / or

[0312] (b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 33; and / or

[0313] (c) VL CDR3 comprising the amino acid sequence of SEQ ID NO:34.

[0314] In specific aspects, the antibody, or antigen-binding portion thereof, comprises one, two, or all three of the above-described VL CDRs.

[0315] In certain aspects, an anti-FAM19A5 antibody or antigen-binding portion thereof that specifically binds to human FAM19A5 comprises:

[0316] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 20;

[0317] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 21;

[0318] (c) a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 22;

[0319] (d) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 32;

[0320] (e) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 33; and / or

[0321] (f) VL CDR3 comprising the amino acid sequence of SEQ ID NO:34.

[0322] In specific aspects, an anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises one, two, three, four, five or six of the above-mentioned CDRs.

[0323] The VH domains described herein or one or more of their CDRs can be linked to a constant domain to form a heavy chain, such as a full-length heavy chain. Similarly, the VL domains described herein or one or more of their CDRs can be linked to a constant domain to form a light chain, such as a full-length light chain. The full-length heavy chain and the full-length light chain are combined to form a full-length antibody.

[0324] Therefore, in a specific aspect, antibodies comprising antibody light chains and heavy chains (e.g., separate light chains and heavy chains) are provided herein. Regarding light chains, in a specific aspect, the light chains of antibodies described herein are κ light chains. In another specific aspect, the light chains of antibodies described herein are λ light chains. In another specific aspect, the light chains of antibodies described herein are human κ light chains or human λ light chains. In a particular embodiment, antibodies described herein that specifically bind to FAM19A5 polypeptides (e.g., human FAM19A5) include light chains, the light chains include any VL or VLCDR amino acid sequences described herein, and the constant region of the light chain includes the amino acid sequence of the human κ light chain constant region. In a specific aspect, antibodies described herein that specifically bind to FAM19A5 polypeptides (e.g., human FAM19A5) include light chains, the light chains include VL or VLCDR amino acid sequences described herein, wherein the constant region of the light chain includes the amino acid sequence of the human λ light chain constant region. Non-limiting examples of human constant region sequences are described in the art, eg, see US Pat. No. 5,693,780 and Kabat EA et al., (1991) supra.

[0325] Regarding the heavy chain, in certain aspects, the heavy chain of the antibody described herein may be an α (a), δ (d), ε (e), γ (g) or μ (m) heavy chain. In another specific aspect, the heavy chain of the antibody may include human α (a), δ (d), ε (e), γ (g) or μ (m) heavy chain. In one aspect, the antibody described herein that specifically binds to FAM19A5 (e.g., human FAM19A5) includes a heavy chain, the heavy chain includes a VH or VHCDR amino acid sequence described herein, wherein the constant region of the heavy chain includes the amino acid sequence of the human γ (g) heavy chain constant region. In another embodiment, the antibody described herein that specifically binds to FAM19A5 (e.g., human FAM19A5) includes a heavy chain, the heavy chain includes a VH or VHCDR amino acid sequence disclosed herein, wherein the constant region of the heavy chain includes the amino acid of a human heavy chain described herein or known in the art. Non-limiting examples of human constant region sequences are described in the art, eg, see US Pat. No. 5,693,780 and Kabat EA et al., (1991) supra.

[0326] In certain aspects, antibodies described herein that specifically bind to FAM19A5 (e.g., human FAM19A5) include VL domains and VH domains, including VH or VHCDR and VL and VLCDR as described herein, wherein the constant region includes the amino acid sequence of the constant region of IgG, IgE, IgM, IgD, IgA or IgY immunoglobulin molecules or human IgG, IgE, IgM, IgD, IgA or IgY immunoglobulin molecules. In another specific aspect, antibodies described herein that specifically bind to FAM19A5 (e.g., human FAM19A5) include VL domains and VH domains, including any amino acid sequence described herein, wherein the constant region includes the amino acid sequence of the constant region of IgG, IgE, IgM, IgD, IgA or IgY immunoglobulin molecules, any subclass of immunoglobulin molecules (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2). In certain aspects, the constant region comprises the amino acid sequence of a naturally occurring human IgG constant region, including subclasses (e.g., IgG1, IgG2, IgG3, or IgG4) and allotypes (e.g., G1m, G2m, G3m, and nG4m) and variants thereof. For example, see Vidarsson G. et al., Front Immunol. 5:520 (published online on October 20, 2014) and Jefferis R. and Lefranc MP, Monoclonal Antibodies (mAbs) 1:4, 1-7 (2009). In certain aspects, the constant region comprises the amino acid sequence of a constant region of human IgG1, IgG2, IgG3, or IgG4, or a variant thereof.

[0327] In certain aspects, the anti-FAM19A5 antibodies disclosed herein or their antigen-binding portions do not have Fc effector functions, such as complement-dependent cytotoxicity (CDC) and / or antibody-dependent cellular phagocytosis (ADCP). The effector functions are mediated by the Fc region, and the residues closest to the hinge region in the CH2 domain of the Fc region are responsible for the effector functions of the antibody because it contains a large portion of overlapping binding sites for C1q (complement) and IgG-Fc receptors (FcγR) on effector cells of the innate immune system. At the same time, the Fc effector function levels of IgG2 and IgG4 antibodies are lower than those of IgG1 and IgG3 antibodies. The effector function of an antibody can be reduced or avoided by various methods known in the art, including (1) using antibody fragments lacking an Fc region (e.g., such as Fab, F(ab')2, single-chain Fv (scFv), or sdAbs composed of monomeric VH or VL domains); (2) generating aglycosylated antibodies, which can be generated by, for example, deleting or changing the residues to which sugars are attached, enzymatically removing sugars, producing antibodies in cells cultured in the presence of glycosylation inhibitors, or expressing antibodies in cells that are unable to glycosylate proteins (e.g., bacterial host cells, e.g., see U.S. Patent (Publication No.: 20120100140)); (3) using an Fc region from an IgG subclass with reduced effector function (e.g., an Fc region from an IgG2 or IgG4 antibody, or a chimeric Fc region comprising a CH2 domain from an IgG2 or IgG4 antibody, e.g., see U.S. Patent (Publication No.: 20120100140) and Lau et al.). C. et al., Journal of Immunology, 191: 4769-4777 (2013); (4) generating an Fc region with mutations that reduce or eliminate Fc function. For example, see U.S. Patent (Publication No.: 20120100140) and the U.S. and PCT applications cited therein and An et al., Monoclonal Antibodies (mAbs), 1: 6,572-579 (2009).

[0328] Thus, in certain aspects, the anti-FAM19A5 antibodies disclosed herein, or antigen-binding portions thereof, are Fab, Fab', F(ab')2, Fv, single-chain Fv (scFv), or sdAbs consisting of monomeric VH or VL domains. Such antibody fragments are well known in the art and are described above.

[0329] In some aspects, the anti-FAM19A5 antibodies disclosed herein or their antigen-binding portions include an Fc region in which Fc effector function is reduced or there is no Fc effector function. In some aspects, the constant region includes the amino acid sequence of a human IgG2 or IgG4 Fc region, and in some aspects, the anti-FAM19A5 antibody is an IgG2 / IgG4 isotype. In some aspects, the anti-FAM19A5 antibody includes a chimeric Fc region, which includes a CH2 domain of an IgG antibody from an IgG4 isotype and a CH3 domain of an IgG antibody from an IgG1 isotype, or includes a chimeric Fc region from a hinge region from IgG2 and a CH2 region from IgG4, or an Fc region with a mutation that causes Fc function to be reduced or there is no Fc function. Fc regions in which Fc effector function is reduced or there is no Fc effector function include Fc regions known in the art. For example, see Lau C. et al., Journal of Immunology (J.Immunol.) 191: 4769-4777 (2013); An et al., Monoclonal Antibodies (mAbs) 1: 6,572-579 (2009); and U.S. Patent (Publication No.: 20120100140) and U.S. Patents and publications and PCT publications cited therein. Those of ordinary skill in the art can also easily prepare Fc regions with reduced or no Fc effector function.

[0330] V. Nucleic Acid Molecules

[0331] Another aspect described herein relates to one or more nucleic acid molecules encoding any of the antibodies described herein or their antigen-binding portions thereof. The nucleic acid can be present in the form of intact cells, cell lysates, or partially purified or substantially purified. The nucleic acid is "separated" or "substantially purified" when separated from other cellular components or other contaminants (e.g., other cellular nucleic acids (e.g., other chromosomal DNA, e.g., chromosomal DNA connected to naturally isolated DNA) or protein purification using standard techniques (including alkali / SDS treatment, CsCl banding, column chromatography, restriction enzymes, agarose gel electrophoresis, and other techniques well known in the art). See F.Ausubel et al., eds., (1987) Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York. The nucleic acid described herein can be, for example, DNA or RNA, and may or may not contain intron sequences. In some aspects, the nucleic acid is a cDNA molecule.

[0332] Nucleic acids as described herein can be obtained using standard molecular biology techniques. For antibodies expressed by hybridomas (e.g., hybridomas prepared by transgenic mice carrying human immunoglobulin genes as further described below), cDNAs encoding the light and heavy chains of antibodies prepared by hybridomas can be obtained by standard PCR amplification or cDNA cloning techniques. For antibodies obtained from immunoglobulin gene libraries (e.g., using phage display technology), nucleic acids encoding antibodies can be recovered from the libraries.

[0333] Certain nucleic acid molecules described herein are nucleic acid molecules encoding the VH and VL sequences of various anti-FAM19A5 antibodies of the present disclosure. Exemplary DNA sequences encoding the VH sequences of such antibodies are listed in SEQ ID NOs: 43-46 and 177. Exemplary DNA sequences encoding the VL sequences of such antibodies are listed in SEQ ID NOs: 47-50 and 178.

[0334] Table 6: Variable heavy chain polynucleotide sequences

[0335]

[0336]

[0337]

[0338]

[0339] Table 7: Variable light chain polynucleotide sequences

[0340]

[0341]

[0342]

[0343] A method for preparing an anti-FAM19A5 antibody disclosed herein, which may include expressing the relevant heavy and light chains of the antibody in a cell line comprising nucleotide sequences encoding heavy and light chains with signal peptides, such as SEQ ID NOs: 43 and 47, SEQ ID NOs: 44 and 48, SEQ ID NOs: 45 and 49, SEQ ID NOs: 46 and 50, SEQ ID NOs: 177 and 178, respectively. Host cells comprising these nucleotide sequences are included herein.

[0344] Once the DNA fragments encoding the VH and VL fragments are obtained, these DNA fragments can be further manipulated using standard recombinant DNA techniques, for example, the variable region genes are converted into full-length antibody chain genes, Fab fragment genes or scFv genes. In these manipulations, the DNA fragment encoding VL or VH is operably linked to another DNA fragment encoding another protein (such as an antibody constant region or a flexible linker peptide). The term "operably linked" as used in this context is intended to indicate that the two DNA fragments are connected so that the amino acid sequences encoded by the two DNA fragments remain in frame.

[0345] The isolated DNA encoding the VH region can be converted into a full-length heavy chain gene by operably linking the DNA encoding the VH to another DNA molecule encoding the heavy chain constant region (hinge, CH1, CH2 and / or CH3). In the art, the sequences of human heavy chain constant region genes are known (e.g., see Kabat, EA et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No.: 91-3242), and DNA fragments including these regions can be obtained by standard PCR amplification. The heavy chain constant region can be an IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM or IgD constant region, such as an IgG2 and / or IgG4 constant region. For a Fab fragment heavy chain gene, the DNA encoding the VH can be operably linked to another DNA molecule encoding only the heavy chain CH1 constant region.

[0346] By making the DNA encoding VL operably linked to another DNA molecule encoding the light chain constant region CL, the isolated DNA encoding the VL region can be converted into a full-length light chain gene (and a Fab light chain gene). In the art, it is known that there are sequences of human light chain constant region genes (e.g., see Kabat, EA et al. (1991) "Sequences of Proteins of Immunological Interest" Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No.: 91-3242), and by standard PCR amplification, DNA fragments including these regions can be obtained. The light chain constant region can be a kappa or lambda constant region.

[0347] To produce an scFv gene, a DNA fragment encoding VH and VL is operably linked to another fragment encoding a flexible linker peptide (e.g., encoding the amino acid sequence (Gly4-Ser)3) so that the VH and VL sequences can be expressed as a continuous single-chain protein, wherein the VL and VH regions are connected by a flexible linker peptide (e.g., see Bird et al. (1988) Science 242:423-426; Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883; McCafferty et al. (1990) Nature 348:552-554).

[0348] In certain aspects, the disclosure provides a vector comprising an isolated nucleic acid molecule comprising a nucleotide sequence encoding an antibody or an antigen-binding portion thereof. In other aspects, the vector can be used for gene therapy.

[0349] Suitable vectors of the present disclosure include expression vectors, viral vectors and plasmid vectors. In one aspect, the vector is a viral vector.

[0350] As used herein, expression vector refers to any nucleic acid construct, which contains the necessary elements for transcription and translation of the inserted coding sequence, or in the case of RNA viral vectors, when introduced into an appropriate host cell, contains the necessary elements for replication and translation. Expression vectors can include plasmids, phagemids, viruses and derivatives thereof.

[0351] The expression vector of the present disclosure may include a polynucleotide encoding an antibody or antigen-binding portion thereof as described herein. In one aspect, the coding sequence of an antibody or antigen-binding portion thereof is operably connected to an expression control sequence. When two nucleic acid sequences used herein are covalently linked so that each constituent nucleic acid sequence can retain its functionality, the two nucleic acid sequences are operably connected. When a coding sequence and a gene expression control sequence are covalently linked so that the expression or transcription and / or translation of the coding sequence is under the influence or control of the gene expression control sequence, it is said to be operably connected. If a promoter is induced in the 5' gene expression sequence, resulting in transcription of the coding sequence, and if the nature of the connection between the two DNA sequences does not (1) result in the introduction of a frameshift mutation, (2) interfere with the ability of the promoter region to guide the transcription of the coding sequence, or (3) interfere with the ability of the corresponding RNA transcript to be translated into a protein, then it can be said that the two DNA sequences are operably connected. Therefore, if the gene expression sequence can affect the transcription of the coding nucleic acid sequence so that the transcript produced thereby is translated into the desired antibody or antigen-binding portion thereof, the gene expression sequence will be operably connected to the coding nucleic acid sequence.

[0352] Viral vectors include, but are not limited to, nucleic acid sequences from the following viruses: retroviruses, such as Moloney murine leukemia virus, Harvey murine sarcoma virus, mouse mammary tumor virus, and Rous sarcoma virus; lentivirus; adenovirus; adeno-associated virus; SV40 virus; polyoma virus; Epstein-Barr virus; papillomavirus; herpes virus; vaccinia virus; poliovirus; and RNA viruses, such as retroviruses. Other vectors known in the art are easy to use. Certain viral vectors are based on non-cytopathic eukaryotic viruses in which non-essential genes have been replaced with target genes. Non-cytopathic viruses include retroviruses, whose life cycle includes reverse transcription of genomic viral RNA into DNA, followed by proviral integration into host cell DNA. Retroviruses have been approved for human gene therapy trials. The most useful retroviruses are replication-deficient (i.e., capable of directing the synthesis of desired proteins, but unable to make infectious particles) retroviruses. Such genetically modified retroviral expression vectors have the general utility of efficient gene transduction in vivo. Standard protocols for producing replication-defective retroviruses (including the steps of incorporating foreign genetic material into a plasmid, transfecting a packaging cell line with the plasmid, producing recombinant retroviruses by the packaging cell line, collecting viral particles from tissue culture medium, and infecting target cells with the viral particles) are provided in Kriegler, M., Gene Transfer and Expression, A Laboratory Manual, WH Freeman Co., New York (1990) and Murry, EJ, Methods in Molecular Biology, Vol. 7, Humana Press, Inc., Cliffton, New Jersey (1991).

[0353] In one aspect, the virus is an adeno-associated virus, i.e., a double-stranded DNA virus. Adeno-associated virus can be designed as a replication-defective virus and can infect a wide range of cell types and species. It also has advantages such as thermal stability and lipid solvent stability; high transduction frequency in cells of different lineages, including hematopoietic cells; and lack of inhibition by repeated infection, thereby allowing multiple series of transductions. It is reported that adeno-associated virus can integrate into human cell DNA in a site-specific manner, thereby minimizing the possibility of insertional mutagenesis and the variability of inserted gene expression characteristics of retroviral infection. In addition, in the absence of selective pressure, wild-type adeno-associated virus infection has been tracked in tissue culture for more than 100 generations, which means that adeno-associated virus genome integration is a relatively stable event. Adeno-associated virus can also act in an extrachromosomal manner.

[0354] In other aspects, the vector is derived from a lentivirus. In certain aspects, the vector is a recombinant lentiviral vector capable of infecting non-dividing cells.

[0355] Typically, the lentiviral genome and proviral DNA have three genes found in retroviruses: gag, pol, and env, flanked by two long terminal repeat (LTR) sequences. The gag gene encodes internal structural (matrix, capsid, and nucleocapsid) proteins; the pol gene encodes RNA-guided DNA polymerase (reverse transcriptase), protease, and integrase; and the env gene encodes viral envelope glycoproteins. The 5' and 3' LTRs serve to promote transcription and polyadenylation of virion RNA. The LTRs contain all other cis-acting sequences necessary for viral replication. Lentiviruses have additional genes, including vif, vpr, tat, rev, vpu, nef, and vpx (in HIV-1, HIV-2, and / or SIV).

[0356] Adjacent to the 5'LTR are sequences necessary for reverse transcription of the genome (tRNA primer binding site) and efficient encapsidation of viral RNA to form particles (Psi site). If sequences necessary for encapsidation (or packaging of retroviral RNA into infectious virions) are missing from the viral genome, the cis defect will prevent encapsidation of the genomic RNA.

[0357] However, the mutants thus obtained are still able to direct the synthesis of all viral particle proteins. The present disclosure provides a method for producing a recombinant lentivirus capable of infecting non-dividing cells, comprising transfecting a suitable host cell with two or more vectors carrying packaging functions, namely gag, pol and env as well as rev and tat. According to the content to be disclosed below, in certain applications, a vector lacking a functional tat gene is required. Thus, for example, a first vector can provide nucleic acids encoding viral gag and viral pol, and another vector can provide nucleic acids encoding viral env to produce packaging cells. A vector providing a heterologous gene (herein referred to as a transfer vector) is introduced into a packaging cell to produce a production cell, which releases infectious viral particles carrying the target exogenous gene.

[0358] According to the configuration of the vector and foreign gene indicated above, the second vector can provide nucleic acid encoding viral envelope (env) gene. The env gene can be derived from almost any suitable virus, including retroviruses. In some aspects, the env protein is a dual-tropic envelope protein, allowing cell transduction of humans and other species.

[0359] Examples of env genes derived from retroviruses include, but are not limited to, Moloney murine leukemia virus (MoMuLV or MMLV), Harvey murine sarcoma virus (HaMuSV or HSV), mouse mammary tumor virus (MuMTV or MMTV), Gibbon ape leukemia virus (GaLV or GALV), human immunodeficiency virus (HIV), and Rous sarcoma virus (RSV). Other env genes may also be used, such as vesicular stomatitis virus (VSV) protein G (VSV G), as well as env genes from hepatitis viruses and influenza viruses.

[0360] The vector providing the viral env nucleic acid sequence is operably associated with the regulatory sequences described in other paragraphs herein.

[0361] In certain aspects, the vector comprises a lentiviral vector in which the HIV virulence genes env, vif, vpr, vpu and nef are deleted without compromising the ability of the vector to transduce non-dividing cells.

[0362] In certain aspects, the vector comprises a lentiviral vector comprising a deletion of the U3 region of the 3' LTR. The U3 region deletion may be a complete deletion or a partial deletion.

[0363] In certain aspects, the lentiviral vector of the present disclosure comprising the FVIII nucleotide sequence described herein can be transfected in a cell using (a) a first nucleotide sequence comprising gag, pol, or gag and pol genes and (b) a second nucleotide sequence comprising a heterologous env gene; wherein the lentiviral vector lacks a functional tat gene. In other aspects, the cell is further transfected using a fourth nucleotide sequence comprising a rev gene. In certain aspects, the lentiviral vector lacks a functional gene selected from vif, vpr, vpu, vpx, and nef or a combination thereof.

[0364] In certain aspects, the lentiviral vector includes one or more nucleotide sequences encoding a gag protein, a Rev-responsive element, a central polypurine territories (cPPT), or any combination thereof.

[0365] Examples of lentiviral vectors are disclosed in WO9931251, W09712622, W09817815, W09817816 and WO9818934, which are incorporated herein by reference in their entirety.

[0366] Other vectors include plasmid vectors. In the art, plasmid vectors are widely described and are well known to those skilled in the art. For example, see Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory Press, 1989. Studies in the past few years have found that plasmid vectors are particularly advantageous for in vivo delivery of genes to cells because they cannot replicate and integrate into the host genome. However, these plasmids have promoters compatible with host cells and can express peptides from operable coding genes in the plasmids. Some commonly used plasmids are available from commercial suppliers, including pBR322, pUC18, pUC19, various pcDNA plasmids, pRC / CMV, various pCMV plasmids, pSV40 and pBlue Script. Other examples of specific plasmids include 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, California). Those of ordinary skill in the art are already familiar with other plasmids. In addition, plasmids can be custom designed using standard molecular biology techniques to remove and / or add specific DNA segments.

[0367] VI. Antibody Production

[0368] Antibodies or fragments thereof that immunospecifically bind to FAM19A5 (e.g., human FAM19A5) can be produced by any antibody synthesis method known in the art, such as by chemical synthesis or recombinant expression technology. Unless otherwise indicated, the methods described herein employ conventional techniques in molecular biology, microbiology, genetic analysis, recombinant DNA, organic chemistry, biochemistry, PCR, oligonucleotide synthesis and modification, nucleic acid hybridization, and related fields of the art. For example, these techniques are described in the references cited herein and are fully explained in the literature. See, for example, Maniatis T et al., 1982 Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press; Sambrook J et al., (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press; Sambrook J et al., (2001) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA; Ausubel FM et al., Current Protocols in Molecular Biology, John Wiley & Sons (1987 and annual updates); Current Protocols in Immunology, John Wiley & Sons (1987 and annual updates); Gait (ed.) (1984) Oligonucleotide Synthesis: A Practical Method. Eckstein (ed.) (1991) Oligonucleotides and Analogues: A Practical Approach, IRL Press; Birren B et al. (eds.) (1999) Genome Analysis: A Laboratory Manual, Cold Spring Harbor Laboratory Press.

[0369] In a specific aspect, the antibodies described herein refer to antibodies (e.g., recombinant antibodies) prepared, expressed, produced, or isolated by any method involving production, such as by synthesis, genetic engineering of DNA sequences. In certain aspects, such antibodies comprise sequences (e.g., DNA sequences or amino acid sequences) that do not naturally occur within the full antibody germline of an animal or mammal (e.g., a human).

[0370] VII. Pharmaceutical Compositions

[0371] Provided herein are compositions comprising an antibody or antigen-binding portion thereof described herein with a desired degree of purity in a physiologically acceptable carrier, excipient or stabilizer (Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, Pennsylvania, USA). Acceptable carriers, excipients or stabilizers are nontoxic to the recipient at the dosages and concentrations used and include buffers such as phosphates, citrates and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride, benzalkonium chloride; benzethonium chloride; phenol, butyl alcohol or benzyl alcohol; alkyl parabens such as methyl paraben or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; m-cresol); low molecular weight ( less than about 10 residues) polypeptides; 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., zinc-protein complexes); and / or nonionic surfactants such as TWEEN®. TM 、PLURONICS TM or polyethylene glycol (PEG).

[0372] In some aspects, the pharmaceutical composition includes an antibody or its antigen binding portion, bispecific molecule or immunoconjugate as described herein, and optionally one or more additional prophylactic or therapeutic agents in a pharmaceutically acceptable carrier. In some aspects, the pharmaceutical composition includes an effective amount of an antibody or its antigen binding portion as described herein, and optionally one or more additional prophylactic or therapeutic agents in a pharmaceutically acceptable carrier. In some aspects, the antibody is the only active ingredient contained in the pharmaceutical composition. The pharmaceutical composition described herein can be used to reduce FAM19A5 activity, thereby treating eye diseases, disorders or conditions, such as glaucoma.

[0373] Pharmaceutically acceptable carriers for parenteral preparations include: aqueous vehicles, non-aqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending agents and dispersants, emulsifiers, polyvalent agents or chelating agents and other pharmaceutically acceptable substances. Examples of aqueous vehicles include: sodium chloride injection, Ringer's injection, isotonic dextrose injection, sterile water injection, dextrose and lactated Ringer's injection. Non-aqueous parenteral vehicles include: fixed oils of plant origin, cottonseed oil, corn oil, sesame oil and peanut oil. Antimicrobial agents of antibacterial or antifungal concentrations can be added to parenteral preparations packaged in multi-dose containers, and the antimicrobial agents include phenol or cresol, mercurial preparations, benzyl alcohol, chlorobutanol, methylparaben and propylparaben, thimerosal, benzalkonium chloride and benzethonium chloride. Isotonic agents include sodium chloride and dextrose. Buffers include phosphates and citrates. Antioxidants include sodium bisulfate. Local anesthetics include procaine hydrochloride. Suspending and dispersing agents include sodium carboxymethylcellulose, hydroxypropyl methylcellulose and polyvinyl pyrrolidone. Emulsifiers include polysorbate 80 ( 80). Polyvalent complexes or chelators of metal ions include EDTA. Pharmaceutical carriers also include ethanol, polyethylene glycol and propylene glycol for water-miscible vehicles; and sodium hydroxide, hydrochloric acid, citric acid or lactic acid for pH adjustment.

[0374] The pharmaceutical compositions disclosed herein are formulated for use in any individual administration route. In certain aspects, the pharmaceutical compositions disclosed herein may be delivered via any route, and the compositions may be delivered to the subfascial, subconjunctival, suprachoroidal, intravitreal, and other sites within the eye. In certain aspects, the pharmaceutical compositions disclosed herein are delivered to an individual via ocular administration. In certain aspects, intraocular administration includes intravitreal administration. Parenteral administration is also contemplated herein, characterized by subcutaneous, intramuscular, or intravenous injection. Injections may be prepared in conventional forms, either as liquid solutions, or as suspensions, solid forms suitable for dissolution or suspension in liquids prior to injection, or as emulsions. Injections, solutions, and emulsions may also contain one or more excipients. For example, suitable excipients include water, saline, dextrose, glycerol, or ethanol. In addition, if desired, the pharmaceutical composition to be administered may also contain a small amount of non-toxic auxiliary substances, such as agents such as wetting agents or emulsifiers, pH buffers, stabilizers, solubilizers, such as sodium acetate, sorbitan laurate, triethanolamine oleate, and cyclodextrins.

[0375] Preparations for parenteral administration of antibodies include sterile solutions ready for injection, sterile dry soluble products (such as lyophilized powders) ready to be combined with solvents before use, including subcutaneous injection tablets, sterile suspensions ready for injection, sterile dry insoluble products ready to be combined with vehicles before use, and sterile emulsions. The solution can be an aqueous solution or a non-aqueous solution.

[0376] If administered intravenously, suitable carriers include physiological saline or phosphate buffered saline (PBS), and solutions containing thickening and solubilizing agents such as glucose, polyethylene glycol and polypropylene glycol and mixtures thereof.

[0377] The topical mixtures comprising antibodies for topical and systemic administration are prepared according to the described steps. The mixtures thus obtained may be solutions, suspensions, emulsions, etc., and may be formulated into creams, gels, ointments, emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes, foams, aerosols, rinses, sprays, suppositories, bandages, skin patches or any other preparations suitable for topical administration.

[0378] The antibodies or antigen-binding portions thereof described herein can be formulated as a topical aerosol for administration, for example, by inhalation (e.g., see U.S. Pat. Nos. 4,044,126, 4,414,209, and 4,364,923, which describe aerosols for delivering steroids useful for treating inflammatory diseases, particularly asthma). These formulations for respiratory administration can be in the form of aerosols or solutions for nebulizers, or as finely divided powders for insufflation (alone or in combination with an inert carrier such as lactose). In this case, in one aspect, the formulation particles are less than 50 microns in diameter, and in one aspect, the formulation particles are less than 10 microns in diameter.

[0379] The antibodies or antigen-binding portions thereof described herein are formulated for topical or local administration, such as topical administration to the skin and mucous membranes, such as in the eye, in the form of gels, creams and lotions, and for ocular or intracisternal or intraspinal administration. Topical administration is contemplated for transdermal delivery, as well as for administration to the eye or mucous membranes, or for inhalation therapy. Nasal solutions of antibodies may also be administered alone or in combination with other pharmaceutically acceptable excipients.

[0380] Those skilled in the art are familiar with transdermal patches, including iontophoresis and electrophoresis devices, which can be used to administer antibodies. For example, such patches are disclosed in U.S. Pat. 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.

[0381] In some aspects, the pharmaceutical composition comprising the antibody or antigen-binding portion thereof described herein is a lyophilized powder that can be reconstituted for administration as a solution, emulsion, and other mixtures. It can also be reconstituted and formulated as a solid or gel. The lyophilized powder is prepared by dissolving the antibody or antigen-binding portion thereof described herein or a pharmaceutically acceptable derivative thereof in a suitable solvent. In some aspects, the lyophilized powder is sterile. The solvent may contain excipients that improve stability, or other pharmacological components of the reconstituted solution prepared from the powder or the powder. Excipients that can be used include, but are not limited to, dextrose, sorbitol, fructose, corn syrup, xylitol, glycerol, glucose, sucrose, or other suitable agents. The solvent may also contain a buffer, such as citrate, sodium phosphate or potassium phosphate, or other such buffers known to those skilled in the art, and in one aspect, its pH is about neutral. Subsequently, the solution is sterile filtered and then lyophilized under standard conditions known to those skilled in the art to provide the desired formulation. In one aspect, the resulting solution will be distributed into vials for lyophilization. Each vial will contain a single dose or multiple doses of the compound. The lyophilized powder can be stored under appropriate conditions, such as about 4°C to room temperature.

[0382] The lyophilized powder is reconstituted with water for injection to provide a formulation for parenteral administration. During reconstitution, the lyophilized powder is added to sterile water or other suitable carrier. The exact amount depends on the compound selected and can be determined empirically.

[0383] Antibodies or antigen-binding portions thereof, bispecific molecules or immunoconjugates described herein, and other compositions provided herein, after formulation, can also be targeted to specific tissues, receptors or other parts of the body of an individual in need of treatment. Those skilled in the art are familiar with many such targeting methods. All such targeting methods for the compositions of the present invention are contemplated herein. For non-limiting examples of targeted approaches, see, e.g., U.S. Pat. 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. In a specific aspect, the anti-FAM19A5 antibodies described herein, or antigen-binding portions thereof, can be used to treat glaucoma and / or reduce, ameliorate, or inhibit glaucoma-associated inflammation.

[0384] Compositions for in vivo administration can be sterile compositions, which can be easily achieved, for example, by filtration through sterile filtration membranes.

[0385] VIII. Kit

[0386] Provided herein are kits comprising one or more antibodies, or antigen binding portions thereof, bispecific molecules, or immunoconjugates thereof as described herein. In a specific aspect, provided herein are pharmaceutical packs or kits comprising one or more containers filled with one or more ingredients of a pharmaceutical composition as described herein, such as one or more antibodies or antigen binding portions thereof as provided herein, and optional instructions for use. As described herein, in certain aspects, the kit comprises a pharmaceutical composition as described herein and any prophylactic or therapeutic agent.

[0387] Example

[0388] Example 1

[0389] Evaluation of retinal potential difference after in vivo administration of anti-FAM19A5 antibody in diabetic mouse model

[0390] Retinopathy

[0391] To begin evaluating the therapeutic effect of anti-FAM19A5 antibodies on diabetic retinopathy, type 2 diabetic db / db mice (B6.BKS(D)-Leprdb / j) (25 weeks old) were selected. Briefly, after the mice were stabilized for 1 week, the mice (7 doses in total, 4 μg / eye every 2 weeks) were intravitreally injected with human IgG control antibody or anti-FAM19A5 antibody (1-65 or 3-2 clone) using a micropump system. Normal (i.e., non-diabetic wild-type) mice were used as controls. Retinal potentials were then assessed using electroretinography (ERG).

[0392] The ERG is a diagnostic test that measures the electrical activity (i.e., retinal potentials) generated by various cells in the retina, including photoreceptors (rods and cones), inner retinal cells (bipolar and amacrine cells), and ganglion cells. In general, when a bright stimulus flashes, the ERG of a healthy eye will show a complex waveform consisting of an initial negative deflection ("A wave"), followed by a B wave with an increasingly steep rise and increasingly rapid peak, which is a superimposed high-frequency oscillation called an "oscillatory potential" (OP). The A wave is dominated by the common response of the rods, while the scotopic B wave is dominated by the response of the rod bipolar neurons. The OP can be seen in the inner plexiform layer, where bipolar cells, amacrine cells, and ganglion cells interact. Wilsey et al., Curr Opin Ophthalmol 27(2): 118-124 (2016). By evaluating these values, the health of the different cells in the retina can be assessed.

[0393] For the evaluation of retinal potentials using ERG, the pupils of the eyes of the animals in the different treatment groups were dilated and topically treated with 2.5% phenylephrine hydrochloride. The animals were then shaved, anesthetized (using ketamine (80 mg / kg) and xylazine (16 mg / kg), followed by 0.5% procaine hydrochloride), and electrodes were attached to the skin, tail, and cornea. The retinal potentials were assessed with flashing white light (0.9 log cd sec / m 2 ) stimulates the retina. When evaluating the electrical activity in the eye of different animals (i.e., the "retinal potential difference"), the values ​​of the A wave amplitude (from the trough of the A wave to the baseline), the B wave amplitude (from the trough of the A wave to the peak of the B wave), and the oscillatory potential (OP) (the amplitude and duration of the small oscillations observed on the rising branch of the B wave) are measured.

[0394] like Figures 1A-1C As shown, mice that received anti-FAM19A5 antibodies (clone 1-65 or 3-2) had improved A waves, B waves, and oscillatory potentials compared to mice that received control IgG antibodies. This suggests that animals that received anti-FAM19A5 antibodies had restored normal retinal function.

[0395] Example 2

[0396] Evaluation of morphological changes in diabetic retinopathy mouse model after anti-FAM19A5 antibody treatment

[0397] To evaluate whether anti-FAM19A5 antibody treatment would lead to changes in retinal morphology, the type 2 diabetic db / db mice of Example 1 were sacrificed and both eyes were removed.

[0398] Once harvested, one eye from each animal was fixed in 10% neutral formaldehyde for one day. Then, the cornea, lens, and sclera were removed, and the remaining eye tissue was washed five times with distilled water for 1 hour while shaking. After the last wash, the distilled water was removed, and the eye tissue was dissolved in 0.1M Tris buffer and incubated in 3% trypsin at 37°C for 2 hours. Next, the retinal vessels were separated from the rest of the retinal tissue, attached to a slide, and stained with periodic acid-Schiff (PAS) to evaluate the retinal microvasculature and obtain characteristic markers of early diabetic retinopathy (e.g., pericyte loss and acellular capillary formation).

[0399] The remaining animal eyes were pre-fixed in 4% glutaraldehyde and then in 10% formaldehyde for 3 days. The eyes were then placed in disposable boxes and embedded in paraffin blocks. The frozen tissues were then sectioned and mounted on slides for further analysis. H&E, TUNEL assays, and IHC analysis were performed to analyze vascular congestion, vascular markers (CD31 and VEGF), and retinal cell protection.

[0400] Figure 2A Microscopic images (100X magnification) of representative areas of the retina of animals belonging to the different treatment groups are provided. In mice receiving anti-FAM19A5 antibodies (clone 1-65 or 3-2), the ratio of endothelial cells to pericytes (E / P ratio) and the frequency and length of acellular capillaries detected were similar to those of normal healthy animals. Figure 2B-2D In contrast, mice treated with human IgG control antibody had a much higher E / P ratio, indicating a loss of pericytes. Figure 2B . Cell-free capillary formation in human IgG control mice ( Figure 2C ) are also more numerous and the acellular capillaries are longer ( Figure 2D ). This result suggests that anti-FAM19A5 antibodies are able to block pericyte loss, protect retinal microvasculature, and prevent the formation of acellular capillaries in a model of diabetic retinopathy.

[0401] In addition, if Figure 3 As shown, vascular congestion within the retinal nerve fiber layer of mice treated with anti-FAM19A5 antibody (bottom two panels) was also greatly reduced compared with mice receiving human IgG control antibody (upper right panel). TUNEL assays showed that apoptotic cells in both the retinal nerve fiber layer and the inner nuclear layer of mice treated with anti-FAM19A5 antibody were reduced compared with the corresponding layers in the retina of mice treated with human IgG control antibody. Figure 4 This suggests that anti-FAM19A5 antibodies are able to protect retinal cells from damage caused by diabetes.

[0402] Finally, if Figure 5 As shown, the nerve fiber layer of mice treated with human IgG control antibody had a lot of CD31 and VEGF staining, indicating the presence of new blood vessels, marking the entry of diabetic retinopathy into a more advanced stage. In contrast, CD31 and VEGF staining was greatly reduced in mice treated with anti-FAM19A5 antibody (clone 1-65 or 3-2). This result shows that anti-FAM19A5 antibody can effectively block new blood vessels in the diabetic retinopathy model.

[0403] Overall, the above data suggest that anti-FAM19A5 antibodies can improve many potential lesions associated with retinopathy, such as diabetic retinopathy, thereby playing a therapeutic role in the disease.

[0404] Example 3

[0405] Evaluation of the effect of anti-FAMI9A5 antibody on choroidal neovascularization in a mouse model of age-related macular degeneration

[0406] To evaluate the effect of anti-FAM19A5 antibody treatment on age-related macular degeneration, a mouse induced choroidal neovascularization (CNV) model was used. More specifically, the CNV model is an animal model of human wet age-related macular degeneration. Briefly, after one week of stabilization of C57BL / 6 mice, the pupils of the animals' eyes were dilated (topical treatment with 2.5% phenylephrine hydrochloride). Then, 2% fluorescein (Sigma) was intraperitoneally administered for treatment and vascular staining. About 3-5 minutes after the injection of fluorescein, the mice were anesthetized and wet AMD-like lesions were induced at four sites of the optic nerve using a Phoenix ophthalmic laser photocoagulation device with a laser (240 mW, duration 70 ms). Figure 6. Thereafter, on the day of CNV induction (i.e., day 0), 3 days and 7 days after CNV induction, mice were given intravitreal injections of human IgG control antibody or anti-FAM19A5 antibody (4 μg / eye). On the 13th day after CNV induction, the extent of choroidal neovascularization was measured using fundus fluorescein angiography (FFA) and Image J program, and the corrected total fluorescence (CTF = integrated density - (selected lesion area x average fluorescence of background reading)) value was calculated. Retinal tomography was performed using image-guided optical coherence tomography (OCT) equipment to measure the extent of neovascular lesions. Neovascular lesions were quantified using Image J program.

[0407] like Fig. 7A As shown, the fluorescence intensity observed at four sites exposed to the laser (indicated by arrows) was similar in both treatment groups after CNV induction and before antibody treatment (day 0, left column). However, at day 13 after CNV induction, the CTF values ​​of mice treated with anti-FAM19A5 antibody were statistically lower than those of mice treated with human IgG control antibody. Figure 7B . Image-guided optical coherence tomography (OCT) (see Figures 8A-8C ) and H&E staining (see Fig. 10B ), confirmed this effect. These results suggest that anti-FAM19A5 antibodies can inhibit and / or reduce laser-induced choroidal neovascularization.

[0408] Example 4

[0409] Evaluation of retinal potential difference after in vivo administration of anti-FAM19A5 antibody in a mouse model of age-related macular degeneration

[0410] To further evaluate the effect of anti-FAM19A5 antibody treatment on age-related macular degeneration, the retinal potential of Example 3 mice was evaluated using electroretinography (ERG). Briefly, the pupils of the eyes of the animals in the different treatment groups were dilated and topically treated with 2.5% phenylephrine hydrochloride. The animals were then anesthetized and electrodes were attached to the skin, tail, and cornea. The retinal potential was assessed using a flashing white light (0.9 log cd sec / m 2 ) stimulates the retina. To assess the electrical activity in the eyes of different animals (i.e., "retinal potential difference"), the B wave value is measured as described in Example 1 above.

[0411] like Fig. 9 As shown, the B-wave retinal potential of CNV-induced mice treated with anti-FAM19A5 antibody was improved compared with that of mice in the human IgG antibody group. This result indicates that the administration of anti-FAM19A5 antibody to CNV-induced mice restored normal retinal function.

[0412] Example 5

[0413] Evaluation of morphological changes after anti-FAM19A5 antibody treatment in a mouse model of age-related macular degeneration

[0414] Next, in order to evaluate whether the administration of anti-FAM19A5 antibodies would cause morphological changes in the retina, the CNV-induced mice of Examples 3 and 4 were killed and their eyes were removed. The eyes were then fixed in 10% neutral formaldehyde for 48 hours. The fixed eyes were then embedded in paraffin blocks for H&E and IHC analysis. The inhibitory effect of anti-FAM19A5 antibodies on choroidal neovascularization shown in Example 3 was confirmed by H&E. Platelet-derived growth factor (PDGF) was expressed by IHC staining. PDGF is known to be secreted by retinal neurons and plays an important role in the formation of astrocytes and choroidal neovascularization.

[0415] like Fig. 10A and 10B As shown, in CNV-induced mice treated with anti-FAMT9A5 antibody, the volume of choroidal neovascularization was significantly reduced at day 13 after CNV induction compared with the human IgG-treated group. This result confirms earlier data (see Example 3) demonstrating the inhibitory effect of anti-FAM19A5 on choroidal neovascularization. Fig.11 As shown, PDGF staining in the nerve fiber layer and ganglion cell layer of the retina of CNV-induced mice treated with anti-FAM19A5 antibody was significantly reduced compared with that in the IgG-treated group.

[0416] Example 6

[0417] Evaluation of the inhibitory effect of anti-FAM19A5 antibodies on drusen formation

[0418] As described above, drusen are tiny yellow or white accumulations of extracellular material (mainly proteins and lipids) that accumulate between the Bruch's membrane and the retinal pigment epithelium (RPE) of the eye. In order to evaluate whether the administration of anti-FAM19A5 antibodies can inhibit the formation of drusen, male APP / PS-l / tau transgenic dementia mice (C57BL / 6; 4 months old) were used. In brief, mice were intravenously injected with anti-FAM19A5 antibodies at a dose of 0.7 mg / kg, twice a week, for a total of 8 weeks. Age-matched non-treated blank wild-type C57BL / 6 and APP / PS-l / tau transgenic mice were used as controls. Eight weeks after the initial antibody administration, the animals were sacrificed and their eyes were removed. Once removed, the eyes were fixed with 4% paraformaldehyde (PFA) at 4°C for 24 hours. Afterwards, the eyes were frozen in a plastic mold containing OCT compound (Lot No. 686794, Sigma, USA). The frozen ocular tissues were then sliced ​​(thickness: 25 μm) using a microtome (Cryostats CM1860, Leica, Germany) at −20° C. The tissue slices were then mounted on glass slides and stained with oligomeric Aβ and Alexa 647 phalloidin to observe whether drusen were formed.

[0419] like Fig.12 As shown, in untreated APP / PS-l / tau transgenic mice, oligomeric Aβ was strongly expressed in the periphery of the visual pigment cells, while drusen (indicated by positive phalloidin staining) were strongly expressed in the periphery of the retinal tissue, cones, and ganglion cell layers. In contrast, in APP / PS-l / tau transgenic mice treated with anti-FAM19A5 antibody, oligomeric Aβ and drusen staining were negligible, similar to that of blank wild-type C57BL / 6 control mice.

[0420] The above results confirmed that anti-FAM19A5 antibody can inhibit the formation of oligomeric Aβ and drusen in the retina (including surrounding tissues).

[0421] Overall, the above data suggest that anti-FAM19A5 antibodies can also improve many potential lesions associated with macular degeneration, such as age-related macular degeneration, thereby playing a therapeutic role in the disease.

[0422] Example 7

[0423] Evaluation of the inhibitory effect of anti-FAM19A5 antibody on diabetic retinopathy after intravenous injection

[0424] To evaluate the therapeutic effect of anti-FAM19A5 antibodies on diabetic retinopathy after intravenous injection, type 2 diabetic db / db mice (such as the mice described in Example 1) were used. Briefly, the animals were stabilized and then treated with one of the following methods: (i) human IgG antibody, (ii) aflibercept ( Regeneron Pharmaceuticals), or (iii) anti-FAM19A5 antibody (clone 1-30). Human IgG antibody and human IgG antibody were used as negative and positive controls, respectively. Blank mice (i.e., animals without diabetic retinopathy) were used as additional controls. Different treatment regimens were administered, injected into the vitreous cavity of mice using a micropump system (a total of 7 doses, 2 μL / eye, once every two weeks), and also intravenously through the tail vein (a total of 7 doses, 100 μL, once every two weeks). The treatment groups are shown in Table 8 below.

[0425] Table 8. Treatment groups

[0426]

[0427] The overall health of the animals is monitored regularly, for example, by measuring blood glucose levels in serum and / or body weight. After the last dose, the mice are euthanized and both eyes of the animals are removed. Once removed, the eyes are prepared on OCT blocks and then fixed for trypsin digestion analysis, hematoxylin-eosin staining (H&E staining), TUNEL analysis, and immunofluorescence staining as described in Example 2, see below for details. Trypsin digestion analysis is performed to assess the microvasculature for pericyte loss and acellular capillary formation (i.e., markers of early diabetic retinopathy). H&E staining, TUNEL analysis, and immunofluorescence staining are performed to analyze vascular congestion, vascular markers (CD31 and VEGF), and retinal cell protection.

[0428] H&E analysis

[0429] After the tissue sections were frozen and attached to the slides (see Example 2), the slides were dried at room temperature for about an hour. The slides were then stained with hematoxylin for 3-5 minutes and cleaned with a hydrochloric acid solution for 30 seconds. The slides were then soaked in an eosin solution for one minute and cleaned again in a hydrochloric acid solution. Next, the slides were cleaned in 80%-100% ethanol for 3 minutes and reacted with carboxylates and xylene for 5 minutes, respectively. The stained slides were observed using a microscope (NanoZoomer 2.0RS, Hamamatsu).

[0430] TUNEL assay

[0431] The slides containing frozen tissue sections (see Example 2) were dried at room temperature for about 1 hour. Once dry, the slides were soaked in 1% paraformaldehyde for 10 minutes and then washed twice with PBS for 5 minutes. Next, the slides were immersed in the equilibration buffer for 10 minutes. The tissue samples were then treated with TdT enzyme in a humidified chamber at 37°C for 2 hours. The slides were then washed four times with PBS for 2 minutes. The stained slides were completely dried at room temperature. Once dry, the tissue samples were stained with DAPI and observed with a fluorescence microscope (LEICA DM 2500).

[0432] Immunofluorescence analysis

[0433] Slides containing frozen tissue sections (see Example 2) were dried at room temperature for about 1 hour. Once dry, tissue samples were treated with a 5% (v / v) Triton X-100 solution in PBS and washed three times with PBS for 5 minutes. Next, the tissues were blocked with normal donkey serum and a 3% (v / v) BSA, 0.5% Triton X-100 solution for 1 hour. Subsequently, tissue staining was performed overnight at 4°C with anti-VEGF and / or anti-CD3 l primary antibodies (diluted 1:300 with a 3% (v / v) BSA and 0.5% triton X-100 solution in PBS). The next day, the samples were washed three times with PBS for 5 minutes. Then, the tissue samples were incubated with secondary antibodies (Alexa Fluor 555 donkey anti-rabbit IgG, diluted 1:500 with a 3% (v / v) BSA and 0.5% triton X-100 solution in PBS) for 1 hour at room temperature. The samples were then washed three times with PBS for 5 min, stained with DAPI, and observed with a fluorescence microscope (LEICA DM 2500).

[0434] result

[0435] As shown in Table 9 (below) and Fig.13A and 13C (Consistent with the data provided in Example 2), the E / P ratio (i.e., the ratio of endothelial cells to pericytes) of animals treated with anti-FAM19A5 antibody (intravenous injection group and intravitreal administration group) was lower than that of animals treated with anti-human IgG alone ("negative control"). Even compared to the positive control group (i.e., animals treated with aflibercept), the effect of anti-FAM19A5 administration on the E / P ratio was greater and similar to the ratio observed in blank (i.e., healthy) animals. The best effect was observed with intravitreal administration, but significant therapeutic effects were also observed with intravenous administration. Similar results were observed when measuring the number of acellular capillaries in the eyes of the animals (see Tables 9 and Fig. 13B).

[0436] Table 9. Histological analysis

[0437] Treatment Group EP Ratio Average AC blank 1.62±0.20 1.78±1.00 Negative control 2.66±0.56 3.36±2.02 Positive Control 2.04±0.33 2.20±1.28 Anti-FAM19A5 antibody (intravitreal administration) 1.38±0.43 1.97±0.97 Anti-FAM19A5 antibody (intravenous injection) 1.66±0.38 1.94±1.17

[0438] The route of administration did not appear to have a significant effect on the ability of anti-FAM19A5 antibodies to reduce vascular congestion and protect retinal cells from damage caused by diabetes (see Fig.14 ). At the same time, Fig.15 As shown, TUNEL assays showed that the number of apoptotic cells in the retinal nerve fiber layer and inner nuclear layer of mice treated with anti-FAM19A5 antibody by intravenous injection was lower than that in the corresponding layers of the retina of mice treated with human IgG control antibody. The effect was close to that of animals treated with anti-FAM19A5 antibody by intravitreal injection (see Fig.14 and 15 Similar results were observed when the expression of CD31 and VEGF (markers of neovascularization) was evaluated. Compared with control animals (i.e., animals treated with human IgG control antibody alone), CD31 and VEGF expression was greatly reduced in both animals treated with anti-FAM19A5 antibody by intravitreal or intravenous injection (see Fig.16A and 16B ).

[0439] Overall, the above data confirm the results of Example 2 (i.e., intravitreal administration) and further demonstrate that anti-FAM19A5 antibodies have therapeutic effects on diabetic retinopathy after both intravenous and intravitreal administration.

[0440] It should be understood that the Detailed Description section, and not the Summary and Abstract sections, is intended to interpret the claims. The Summary and Abstract sections may set forth one or more but not all exemplary aspects of the present disclosure as contemplated by the inventors, and thus are not intended to limit the present disclosure and the appended claims in any way.

[0441] The present disclosure is described above with the aid of functional building blocks, and examples are given of the implementation of the specified functions and their relationships. For ease of description, the boundaries of these functional building blocks are arbitrarily defined herein. Alternative boundaries may be defined as long as the specified functions and their relationships are properly performed.

[0442] The above description of specific aspects will fully reveal the general nature of the present disclosure, so that others can easily modify and / or adapt these specific aspects to various applications by applying knowledge within the technical scope of the art without undue experimentation and without departing from the general concept of the present disclosure. Therefore, based on the teachings and guidance set forth herein, such adaptations and modifications are intended to be within the meaning and scope of equivalents of the various aspects disclosed. It should be understood that the wording or terminology herein is intended to be descriptive rather than limiting, so that the skilled person can interpret the terms or wording of this specification in accordance with the teachings and guidance.

[0443] The breadth and scope of the present disclosure should not be limited by any of the above-described exemplary aspects, but should be defined only in accordance with the following claims and their equivalents.

[0444] 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.

Claims

1. A pharmaceutical composition comprising an antagonist to a family of sequence similarity 19 member A5 ("FAM19A5") protein, i.e., a FAM19A5 antagonist, for preparing a medicament for treating diabetic retinopathy or age-related macular degeneration, The FAM19A5 antagonist is an antibody or an antigen-binding portion thereof that specifically binds to the FAM19A5 protein, i.e., an anti-FAM19A5 antibody, a polynucleotide encoding the anti-FAM19A5 antibody, or a vector comprising the polynucleotide thereof, The anti-FAM19A5 antibody comprises heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3, in, The heavy chain CDR1 comprises the amino acid sequence shown in SEQ ID NO: 212, the heavy chain CDR2 comprises the amino acid sequence shown in SEQ ID NO: 213, the heavy chain CDR3 comprises the amino acid sequence shown in SEQ ID NO: 16, the light chain CDR1 comprises the amino acid sequence shown in SEQ ID NO: 222, the light chain CDR2 comprises the amino acid sequence shown in SEQ ID NO: 225, and the light chain CDR3 comprises the amino acid sequence shown in SEQ ID NO:

224.

2. Use of the pharmaceutical composition according to claim 1 in the preparation of a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The diabetic retinopathy is non-proliferative diabetic retinopathy (NPDR).

3. Use of the pharmaceutical composition according to claim 1 in the preparation of a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The diabetic retinopathy is proliferative diabetic retinopathy.

4. Use of the pharmaceutical composition according to claim 1 in the preparation of a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The age-related macular degeneration refers to dry (atrophic) macular degeneration.

5. Use of the pharmaceutical composition according to claim 1 in preparing a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The age-related macular degeneration refers to wet (neovascular or exudative) macular degeneration.

6. Use of the pharmaceutical composition according to claim 1 in the preparation of a medicament for treating diabetic retinopathy or age-related macular degeneration, wherein: The age-related macular degeneration is (i) early AMD, (ii) intermediate AMD or (iii) advanced AMD (geographic atrophy).

7. Use of the pharmaceutical composition according to claim 1 in the preparation of a medicament for treating diabetic retinopathy or age-related macular degeneration, wherein: Age-related macular degeneration is associated with accumulation or aggregation of drusen, neovascularization, decreased retinal potential, astrogenesis, or any combination thereof within an individual's retina.

8. Use of the pharmaceutical composition according to claim 1 in the preparation of a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The anti-FAM19A5 antibody specifically binds to at least one FAM19A5 epitope, namely, SEQ ID NO:6 or SEQ ID NO:

9.

9. The pharmaceutical composition according to claim 1 is used in the preparation of a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The anti-FAM19A5 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), The VH consists of the amino acid sequence of SEQ ID NO: 236, and the VL consists of the amino acid sequence shown in SEQ ID NO:

245.

10. Use of the pharmaceutical composition according to claim 1 in preparing a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The anti-FAM19A5 antibody includes Fab, Fab', F(ab')2, Fv or single-chain Fv (scFv).

11. Use of the pharmaceutical composition according to claim 1 in preparing a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The anti-FAM19A5 antibody is selected from the group consisting of IgG1, IgG2, IgG3, IgG4 and variants thereof.

12. Use of the pharmaceutical composition according to claim 11 in preparing a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The anti-FAM19A5 antibody is IgG2, IgG4 or a combination thereof.

13. Use of the pharmaceutical composition according to claim 12 in the preparation of a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The anti-FAM19A5 antibodies include IgG2 / IgG4 isotype antibodies.

14. Use of the pharmaceutical composition according to claim 1 in preparing a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The anti-FAM19A5 antibody includes a constant region without Fc function.

15. Use of the pharmaceutical composition according to claim 1 in preparing a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The anti-FAM19A5 antibody is a chimeric antibody, a human antibody or a humanized antibody.

16. Use of the pharmaceutical composition according to claim 1 in preparing a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The anti-FAM19A5 antibody is linked to a molecule having a second binding moiety, thereby forming a bispecific molecule.

17. Use of the pharmaceutical composition according to claim 1 in preparing a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The anti-FAM19A5 antibody is linked to a reagent, thereby forming an immunoconjugate.

18. Use of the pharmaceutical composition according to claim 1 in preparing a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: Further included are therapeutic agents.

19. Use of the pharmaceutical composition according to claim 1 in preparing a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The pharmaceutical composition is formulated with a pharmaceutically acceptable carrier.

20. Use of the pharmaceutical composition according to claim 1 in preparing a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The pharmaceutical composition is administered intraocularly.

21. Use of the pharmaceutical composition according to claim 20 in preparing a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: Intraocular administration includes intravitreal administration.

22. Use of the pharmaceutical composition according to claim 1 in preparing a drug for treating diabetic retinopathy or age-related macular degeneration, wherein: The pharmaceutical composition is administered intravenously.

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