Use of antibodies against sequence-similar family 19 member A5 in treatment of neuropathic pain

Abstract

The present disclosure provides a method for treating neuropathic pain, including central neuropathic pain or peripheral neuropathic pain, in a subject comprising administering antibodies that specifically bind to human FAM19A5 and compositions comprising such antibodies. The present disclosure also provides methods for treating neuropathic pain, including symptoms of neuropathic pain and / or a root cause of neuropathic pain, by administering an antibody that specifically binds to human FAM19A5. In one specific aspect, the method comprises administering an antibody that specifically binds to human FAM19A5, such as to soluble human FAM19A5 at a KD of 10 nM or less, or to membrane-bound human FAM19A5 at a KD of 10 nM or less, as measured by methods known in the art, such as ELISA, or both.

Description

[0001] Reference to Electronically Submitted Sequence Listings

[0002] The contents of the electronically submitted sequence listing in the ASCII text file submitted with this application (Name: 3763_009PC01_SeqListing_ST25.txt; Size: 214,313 bytes; Creation Date: April 22, 2019) are incorporated herein by reference in their entirety. Technical Field

[0003] The present disclosure provides methods for treating neuropathic pain in a subject (eg, a human) using an antibody or antigen-binding fragment thereof that specifically binds to family of sequence similarity 19, member A5 (FAM19A5), or a composition comprising such an antibody or antigen-binding fragment thereof. Background Art

[0004] Neuropathic pain is a chronic pain caused by damage or disease of the somatosensory system. Colloca L et al., Nat Rev Dis Primers 3:17002 (2017). The somatosensory system allows the perception of touch, pressure, pain, temperature, position, movement and vibration. Somatosensory nerves appear in the skin, muscles, joints and fascia, and include thermoreceptors, mechanoreceptors, chemoreceptors, itch receptors and nociceptors, which send signals to the spinal cord and eventually to the brain for further processing. Damage or disease of the somatosensory nervous system can cause changes and disturbances in the transmission of sensory signals to the spinal cord and brain, leading to chronic pain.

[0005] Neuropathic pain places a heavy burden on patients, society, and the healthcare system. It is estimated that approximately 7-10% of the general population suffers from neuropathic pain. Cruccu G. and Truini A., Pain Ther 6:35-42 (2017). The prevalence of neuropathic pain may increase further in the future due to various factors (e.g., aging population, increased obesity rates, increased survival rates of cancer patients receiving interventional treatments that may cause neuropathic pain). Moulin D et al., Pain Res Manag 19:328-335 (2014).

[0006] Because the root cause is not always completely clear, the management of neuropathic pain is often complex and remains a challenge. Finnerup NB et al., Lancet Neurol 14: 162-173 (2015). Current methods focus only on treating the symptoms of neuropathic pain with pharmacological agents such as tricyclic antidepressants (e.g., amitriptyline, serotonin-norepinephrine reuptake inhibitors (SNRIs), and calcium channel α-2-δ ligands gabapentin and pregabalin). Cruccu G and Truini A., Pain Ther 6: 35-42 (2017). Such drugs usually have serious side effects and / or limited efficacy for many individuals. Therefore, more effective treatment options for neuropathic pain are currently needed. Summary of the invention

[0007] Disclosed herein is a method for treating neuropathic pain (including central neuropathic pain or peripheral neuropathic pain) in a subject in need thereof. In some embodiments, the method comprises administering to the subject an antagonist of a family of sequence similarity 19 member A5 (FAM19A5) protein.

[0008] In some embodiments, the neuropathic pain is associated with physical injury, infection, diabetes, cancer therapy, alcoholism, amputation, muscle weakness in the back, legs, buttocks or face, trigeminal neuralgia, multiple sclerosis, herpes zoster, spinal surgery, or any combination thereof. In some embodiments, the neuropathic pain comprises carpal tunnel syndrome, central pain syndrome, degenerative disc disease, diabetic neuropathy, phantom limb pain, postherpetic neuralgia (shingles), pudendal neuralgia, sciatica, low back pain, trigeminal neuralgia, or any combination thereof.

[0009] In some embodiments, the neuropathic pain is caused by nerve compression. In some embodiments, the diabetic neuropathy is diabetic peripheral neuropathy. In some embodiments, the neuropathic pain is sciatica.

[0010] Also provided herein is a method of increasing the threshold or latency of an external stimulus in a subject in need thereof, comprising administering to the subject an antagonist of an anti-FAM19A5 protein. In some embodiments, the external stimulus is a mechanical stimulus. In other embodiments, the external stimulus is a thermal stimulus.

[0011] The present disclosure also provides a method of increasing sensory nerve conduction velocity in a subject in need thereof, comprising administering an antagonist against the FAM19A5 protein to the subject.

[0012] In some embodiments, the FAM19A5 antagonist is an antisense oligonucleotide, siRNA, shRNA, miRNA, dsRNA, aptamer, PNA or a vector comprising them that specifically targets FAM19A5. In other embodiments, the FAM19A5 antagonist is an anti-FAM19A5 antibody, a polynucleotide encoding an anti-FAM19A5 antibody, a vector comprising the polynucleotide thereof, a cell comprising the polynucleotide thereof or any combination thereof.

[0013] In some embodiments, the FAM19A5 antagonist is an anti-FAM19A5 antibody.

[0014] In some embodiments, the anti-FAM19A5 antibody exhibits a property selected from the following: (a) binding to soluble human FAM19A5 with a KD of 10 nM or less as measured by enzyme-linked immunosorbent assay (ELISA); (b) binding to membrane-bound human FAM19A5 with a KD of 10 nM or less as measured by ELISA; or (c) both (a) and (b).

[0015] In some embodiments, the anti-FAM19A5 antibody cross-competes for binding to a human FAM19A5 epitope with 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 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; (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: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; or (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.

[0016] In some embodiments, the anti-FAM19A5 antibody binds to the same FAM19A5 epitope as 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 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; (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: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; or (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.

[0017] In some embodiments, the anti-FAM19A5 antibody binds to at least one FAM19A5 epitope that is SEQ ID NO: 6 or SEQ ID NO: 9. In other embodiments, the anti-FAM19A5 antibody binds only to a FAM19A5 epitope that is SEQ ID NO: 6 or SEQ ID NO: 9. In certain embodiments, the anti-FAM19A5 antibody further binds to an additional FAM19A5 epitope. In some embodiments, the additional FAM19A5 epitope is selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and any combination thereof.

[0018] In some embodiments, the anti-FAM19A5 antibody comprises a heavy chain CDR1, CDR2 and CDR3 and a light chain CDR1, CDR2 and CDR3. In certain embodiments, the heavy chain CDR3 comprises an amino acid sequence of SEQ ID NO: 19, SEQ ID NO: 16, SEQ ID NO: 13 or SEQ ID NO: 22. In other embodiments, the heavy chain CDR1 comprises an amino acid sequence of SEQ ID NO: 17, SEQ ID NO: 14, SEQ ID NO: 11 or SEQ ID NO: 20. In some embodiments, the heavy chain CDR2 comprises an amino acid sequence of SEQ ID NO: 18, SEQ ID NO: 15, SEQ ID NO: 12 or SEQ ID NO: 21.

[0019] In some embodiments, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 29, SEQ ID NO: 26, SEQ ID NO: 23, or SEQ ID NO: 32. In certain embodiments, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 30, SEQ ID NO: 27, SEQ ID NO: 24, or SEQ ID NO: 33. In some embodiments, the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 31, SEQ ID NO: 28, SEQ ID NO: 25, or SEQ ID NO: 34.

[0020] In some embodiments, the anti-FAM19A5 antibody comprises a heavy chain CDR1, CDR2 and CDR3 and a light chain CDR1, CDR2 and CDR3, wherein (i) the heavy chain CDR1, CDR2 and CDR3 comprise SEQ ID NOs: 17, 18 and 19, respectively, and the light chain CDR1, CDR2 and CDR3 comprise SEQ ID NOs: 29, 30 and 31, respectively; (ii) the heavy chain CDR1, CDR2 and CDR3 comprise SEQ ID NOs: 14, 15 and 16, respectively, and the light chain CDR1, CDR2 and CDR3 comprise SEQ ID NOs: 26, 27 and 28, respectively; (iii) the heavy chain CDR1, CDR2 and CDR3 comprise SEQ ID NOs: 11, 12 and 13, respectively, and the light chain CDR1, CDR2 and CDR3 comprise SEQ ID NOs: 23, 24 and 25, respectively; or (iv) the heavy chain CDR1, CDR2 and CDR3 comprise SEQ ID NOs: ID NOs: 20, 21 and 22, and the light chain CDR1, CDR2 and CDR3 comprise SEQ ID NOs: 32, 33 and 34, respectively.

[0021] In some embodiments, the anti-FAM19A5 antibody comprises a heavy chain variable domain comprising SEQ ID NO: 37, SEQ ID NO: 36, SEQ ID NO: 35, or SEQ ID NO: 38 and / or a light chain variable domain comprising SEQ ID NO: 41, SEQ ID NO: 40, SEQ ID NO: 39, or SEQ ID NO: 42. In certain embodiments, the anti-FAM19A5 antibody comprises a heavy chain variable domain comprising SEQ ID NO: 37 and a light chain variable domain comprising SEQ ID NO: 41. In other embodiments, the anti-FAM19A5 antibody comprises a heavy chain variable domain comprising SEQ ID NO: 36 and a light chain variable domain comprising SEQ ID NO: 40. In some embodiments, the anti-FAM19A5 antibody comprises a heavy chain variable domain comprising SEQ ID NO: 35 and a light chain variable domain comprising SEQ ID NO: 39. In certain embodiments, an anti-FAM19A5 antibody comprises a heavy chain variable domain comprising SEQ ID NO:38 and a light chain variable domain comprising SEQ ID NO:42.

[0022] In some embodiments, the anti-FAM19A5 antibody comprises 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: 37, 36, 35, or 38. In some embodiments, the anti-FAM19A5 antibody comprises 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 set forth in SEQ ID NO: 41, 40, 39, or 42. In some embodiments, the anti-FAM19A5 antibody comprises 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 shown in SEQ ID NO:37, 36, 35 or 38; and 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 shown in SEQ ID NO:41, 40, 39 or 42.

[0023] In some embodiments, the anti-FAM19A5 antibody comprises Fab, Fab', F(ab')2, Fv, or single-chain Fv (scFv).

[0024] In some embodiments, the anti-FAM19A5 antibody is selected from the group consisting of: IgG1, IgG2, IgG3, IgG4, variants thereof, and any combination thereof. In certain embodiments, the anti-FAM19A5 antibody is an IgG2 antibody, an IgG4 antibody, or a combination thereof. In other embodiments, the anti-FAM19A5 antibody comprises an IgG2 / IgG4 isotype antibody.

[0025] In some embodiments, the anti-FAM19A5 antibody further comprises a constant region without Fc function. In some embodiments, the anti-FAM19A5 antibody is a chimeric antibody, a human antibody, or a humanized antibody.

[0026] In some embodiments, the FAM19A5 antagonist is linked to an agent to form an immunoconjugate. In some embodiments, the FAM19A5 antagonist is formulated with a pharmaceutically acceptable carrier. In some embodiments, the FAM19A5 antagonist is administered intravenously, orally, parenterally, intrathecally, intracerebroventricularly, intrapulmonaryly, intramuscularly, subcutaneously, intraperitoneally, intravitreally, or intraventricularly.

[0027] In some embodiments, the subject is a human. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1A , 1B 1C shows the binding analysis of single scFv clones to FAM19A5 protein. The absorbance was measured at 405 nM. The clone number is shown on the X-axis. Figure 1A , 1B 1A and 1C show the analysis of 96 clones from level 3, 4 or 5 biopanning of the first, second and third chickens, respectively.

[0029] Figure 2 A schematic diagram for subcloning anti-FAM19A5 antibody (scFv) into a mammalian expression vector is shown.

[0030] Figure 3 The SDS-PAGE results of chimeric anti-FAM19A5-IgG2 / 4 monoclonal antibody (1-65) are shown. The left figure shows reducing SDS-PAGE, and the right figure shows non-reducing SDS-PAGE.

[0031] Figure 4A It was shown that the chimeric anti-FAM19A5-IgG2 / 4 monoclonal antibody (1-65) specifically binds to human FAM19A5.

[0032] Figure 4B It was shown that the chimeric anti-FAM19A5-IgG2 / 4 monoclonal antibody (1-65) is specific for the FAM19A5 protein but does not bind to other proteins in the FAM19A subfamily.

[0033] Figure 5A and 5BIt is shown that applying anti-FAM19A5 antibody alleviates the mechanical hyperalgesia of rats induced by chronic contraction injury (CCI). The impact on mechanical hyperalgesia is shown as a threshold value, under which animals withdraw their claws in response to mechanical stimulation. Before threshold measurement, animals have received one of the following: (i) control antibody (human IgG) (" G2 "), (ii) anti-FAM19A5 antibody (" G3 ") or (iii) pregabalin (" G4 "). Initial (healthy) animals are used as "sham control" (that is, without CCI induction and without antibody administration) (" G1 "). Figure 5A Threshold data on day 6 after CCI induction are shown. Figure 5B Threshold data at day 21 after CCI induction are shown. Figure 5A and 5B In the figure, the bar graphs show the data as mean ± SD. The table below the graph shows the data from individual mice in each group.

[0034] Fig. 6A and 6B It is shown that the use of anti-FAM19A5 antibodies improves the motor function of rats induced by CCI. As described in the examples, the effect on motor function is shown as the latency (seconds) of falling from the rotarod instrument. Before threshold measurement, the animals induced by CCI have received one of the following: (i) control antibody (human IgG) ("G2"), (ii) anti-FAM19A5 antibody ("G3") or (iii) pregabalin ("G4"). The initial (healthy) animals are used as "false controls" (i.e., without CCI induction and without antibody administration) ("G1"). Fig. 6A Data from day 6 after CCI induction are shown. Figure 6B Data from day 21 after CCI induction are shown. The bar graph shows latency data as mean ± SD. The table below the graph shows data from individual mice in each group.

[0035] Figure 7 An alignment of the amino acid sequences of FAM19A5 of different species (i.e., human, mouse, rat, and chicken) is shown. Fragments F1-F6 used for epitope mapping analysis are shown. The signal peptide is underlined.

[0036] Figure 8 The amino acid sequence of epitope F1-F6 (conjugated with BS) and its position on human FAM19A5 polypeptide are shown. The top amino acid sequence shown is wild-type FAM19A5 isoform 2 (without signal peptide). The second amino acid sequence shown is the same sequence, but the cysteine ​​residue is mutated to serine to reduce the non-specific activity during peptide synthesis. The size of different epitope fragments is shown in brackets.

[0037] Fig. 9 The Western blot results for the epitope mapping of monoclonal antibody clone 1-65 and epitope fragments F1 to F6 (swimming lanes 3-8, respectively) are shown. FAM19A5-Cκ (swimming lane 1), PSA-Cκ (swimming lane 2), peptide NDV-BSA (swimming lane 9) and BSA (swimming lane 10) are used as controls. The respective sizes of the different antigens used are shown on the right side of the blot. The amount of the antigen used in each well is 300ng. The primary antibody used for Western blot is 1-65-scFv-rabbit-Fc-SSS (2 μg / mL), and the secondary antibody used for experiment is anti-rabbit IgG (Fc specificity)-HRP (1:4000).

[0038] Fig.10 It is shown that the administration of anti-FAM19A5 antibodies improves the mechanical allodynia of diabetic peripheral neuropathy (DPN) rats. The effect on mechanical allodynia is shown as a threshold (g), at which animals withdraw their paws in response to mechanical stimulation ("paw withdrawal threshold") . About three weeks after inducing diabetic peripheral neuropathy in rats, animals received administration of control antibodies (NHI, open squares, n = 9) or anti-FAM19A5 antibodies (3-2) (solid squares, n = 9) weekly. Initial (i.e. healthy, no induction of DPN) animals were used as controls (open circles, n = 5). Data are expressed as mean ± SD. "*" indicates a statistically significant difference (p < 0.05) compared to animals receiving control antibodies.

[0039] Fig.11A and 11B It is shown that the administration of anti-FAM19A5 antibody improves thermal hyperalgesia in diabetic peripheral neuropathy (DPN) rats. The effect on thermal hyperalgesia is shown as the duration before the animals withdraw their paws in response to thermal stimulation ("paw withdrawal latency"). DPN rats received control antibody (NHI, squares) or anti-FAM19A5 antibody (3-2) (triangles). Fig.11A Thermal hyperalgesia data are shown 4 weeks after antibody administration. Fig. 11B Thermal hyperalgesia data 8 weeks after antibody administration are shown. Naive (ie healthy, no DPN induction) animals were used as controls. Each symbol represents an individual animal. Mean ± SD is also shown.

[0040] Fig.12It is shown that the administration of anti-FAM19A5 antibodies improves sensory nerve conduction velocity (m / s) in diabetic peripheral neuropathy (DPN) rats 8 weeks after antibody administration. DPN rats receive control antibodies (NHI, squares) or anti-FAM19A5 antibodies (3-2) (triangles). Initial (i.e. healthy, no DPN induction) animals are used as controls. Each symbol represents a separate animal. Mean ± SD is also shown. DETAILED DESCRIPTION

[0041] Disclosed herein is a method for treating neuropathic pain in a subject in need thereof, the method comprising administering to the subject an antagonist (e.g., an isolated monoclonal antibody or an antigen-binding portion thereof) that specifically binds to human family of sequence similarity 19, member A5 (FAM19A5) protein.

[0042] To facilitate understanding of the disclosure provided herein, various terms and phrases are defined. Additional definitions are set forth throughout the detailed description.

[0043] I. Definitions

[0044] Throughout this disclosure, the term "a" or "an" entity refers to one or more of that entity; for example, "antibody" is understood to represent one or more antibodies. Thus, the terms "a" (or "a"), "one or more" and "at least one" can be used interchangeably herein.

[0045] Furthermore, “and / or” as used herein should be considered as a specific disclosure of each of the two specified features or components, with or without the other. Thus, the term “and / or” as used herein in phrases such as “A and / or B” is intended to include “A and B,” “A or B,” “A” (alone), and “B” (alone). Similarly, the term “and / or” as used in phrases such as “A, B, and / or C” is intended to cover each of the following: 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).

[0046] It should be understood that wherever aspects are described with the word "comprising," similar aspects described with the terminology "consisting of" and / or "consisting essentially of" are also provided.

[0047] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present disclosure relates. For example, Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd ed., 1999, Academic Press; and Oxford Dictionary Of Biochemistry And Molecular Biology, Revised, 2000, Oxford University Press provide a general dictionary of many of the terms used in the present disclosure.

[0048] Units, prefixes and symbols are expressed in the form accepted by the International System of Units (SI). Numerical ranges include the numbers that define the range. Unless otherwise indicated, amino acid sequences are written from left to right in the direction of amino to carboxyl. The titles provided herein are not limitations on the various aspects of the disclosure, which can be obtained by reference to the specification as a whole. Therefore, the terms defined immediately below are more fully defined by reference to the entire specification.

[0049] The term "about" is used herein to refer to approximately, roughly, approximately, or in the region of ... When the term "about" is used in conjunction with a numerical range, it modifies the range by extending the boundaries above and below the numerical values ​​recited. Typically, the term "about" can modify a numerical value above and below the recited value by, for example, a variance of 10%, up or down (higher or lower).

[0050] The term "neuropathic pain" refers to pain at any level affecting the central nervous system (CNS) and / or peripheral nervous system due to injury, damage and / or inappropriate function. The term "neuropathic pain" includes any and all types of neuropathic pain, regardless of the cause of the neuropathic pain and any and all symptoms.

[0051] Neuropathic pain includes central neuropathic pain and peripheral neuropathic pain. As used herein, the term "central neuropathic pain" refers to pain caused by disorders, congenital defects, or damage to the central nervous system (i.e., brain or spinal cord). As used herein, the term "peripheral neuropathic pain" refers to pain caused by damage or infection of peripheral sensory nerves.

[0052] Symptoms of neuropathic pain can include persistent / chronic pain, spontaneous pain, and allodynia (e.g., a painful response to a stimulus that is not normally painful), hyperalgesia (e.g., an aggravated response to a painful stimulus that normally causes only mild discomfort, such as a pinprick), hyperesthesia (e.g., an excessive physical sensitivity to stimuli, especially of the skin), or hyperalgesia (e.g., where transient discomfort becomes prolonged, severe pain). In some embodiments, symptoms can be persistent and persist after resolution of the primary cause, if present. Merck Manual, Neuropathic Pain, available at merckmanuals.com / professional / neurologic-disorders / pain / neuropathic-pain; Campbell JN and Meyer R Neuron 52(1):77–92 (2006).

[0053] In some embodiments, types of neuropathic pain may include: (1) neuropathic pain, (2) deafferentation pain syndrome, (3) complex regional pain syndrome (CRPS), and (4) neuropathy (central or peripheral).

[0054] In some embodiments, neuropathic pain is neuralgia, which refers to pain radiating along one or more specific nerves (e.g., cranial nerves), usually without any demonstrable pathological changes in the neural structure.Neuropathy includes, but is not limited to, trigeminal neuralgia (TN), atypical trigeminal neuralgia (ATN), occipital neuralgia, glossopharyngeal neuralgia, postherpetic neuralgia (caused by herpes zoster or herpes), peripheral nerve injury pain, sciatica, lower back pain and atypical facial pain.Chemical irritation, chronic kidney disease, diabetes, inflammation, trauma (including surgery), compression of nerves by nearby structures (e.g., tumors), certain drugs (e.g., cisplatin, paclitaxel or vincristine), porphyria (blood disorders) and infection (e.g., herpes zoster), HIV / AIDS, Lyme disease or syphilis) may all cause neuralgia.

[0055] In some embodiments, neuropathic pain is deafferentation pain syndrome, which can be caused by loss of sensory input from a part of the body (e.g., caused by interruption of peripheral sensory fibers or nerves from the central nervous system). Deafferentation pain syndrome includes, but is not limited to, brain injury or spinal cord injury, post-stroke pain, phantom limb pain, paraplegia, brachial plexus avulsion injury, and lumbar radiculopathy.

[0056] In some embodiments, neuropathic pain is "complex regional pain syndrome" (CRPS), which is a chronic pain condition that most often affects the arms or legs. In some embodiments, CRPS develops after injury, surgery, stroke, or heart attack. In certain embodiments, CRPS disease is type I CRPS (CRPS-I) (also known as reflex sympathetic dystrophy syndrome). Individuals without confirmed nerve damage are generally classified as CRPS-I types. In other embodiments, CRPS is type II CRPS (CRPS-II) (also known as causalgia), which is associated with confirmed nerve damage.

[0057] In some embodiments, neuropathic pain is neuropathy, which refers to the pain caused by the function or pathological changes (for example, disease or injury) of nerves.Neuropathy is usually characterized by sensory or motor neuron abnormalities clinically.In certain embodiments, neuropathy is central neuropathy (for example, the function or pathological changes of the central nervous system).In other embodiments, neuropathy is peripheral neuropathy (for example, the function or pathological changes of one or more peripheral nerves, including motor nerves, sensory nerves, autonomic nerves or their combinations).In some embodiments, peripheral neuropathy relates to the function or pathological changes of a single nerve or nerve group (that is, mononeuropathy).In some embodiments, peripheral neuropathy relates to the function or pathological changes (i.e. polyneuropathy) affecting multiple nerves (local or systemic).In some embodiments, peripheral neuropathy is roughly the same (i.e. symmetrical polyneuropathy) on both sides of the body.In some embodiments, peripheral neuropathy affects different areas of the body (for example, multiple mononeuritis, multifocal mononeuropathy or multiple mononeuropathy).

[0058] As used herein, "mononeuropathy" is a peripheral neuropathy, which relates to the loss of movement or sensation in the area caused by the damage or destruction of a single peripheral nerve or a single nerve group.Mononeuropathy is most often caused by damage or trauma in a local area, for example, it causes long-term pressure / compression on a single nerve. However, some systemic disorders (e.g., multiple mononeuritis) can also cause mononeuropathy. In certain embodiments, damage or trauma in a local area causes the destruction of the myelin sheath (covering) or part of the nerve cells (axons) of the nerve, which can slow down or prevent the conduction of impulses through the nerve. In certain embodiments, mononeuropathy can affect any part of the body. The example of mononeuropathy pain includes but is not limited to sciatic nerve dysfunction, common peroneal nerve dysfunction, radial nerve dysfunction, ulnar nerve dysfunction, cranial mononeuropathy VI, cranial mononeuropathy VII, cranial mononeuropathy III (compression type), cranial mononeuropathy III (diabetic type), axillary nerve dysfunction, carpal tunnel syndrome, femoral nerve dysfunction, tibial nerve dysfunction, Bell's palsy, thoracic outlet syndrome, carpal tunnel syndrome and sixth (abducens) nerve palsy. Finnerup NB et al., Pain 157(8):1599-1606 (2016); National Institute of Neurological Disorders and Stroke, Peripheral Neuropathy Fact Sheet, available at ninds.nih.gov / disorders / peripheralneuropathy / detail_peripheralneuropathy.htm. In some embodiments, the mononeuropathic pain is sciatica.

[0059] As used herein, "polyneuropathy" is a peripheral neuropathy that involves loss of movement or sensation in an area caused by damage or destruction of multiple peripheral nerves. Polyneuropathy pain includes, but is not limited to, post-polio syndrome, post-mastectomy syndrome, diabetic neuropathy, alcoholic neuropathy, amyloid, toxins, AIDS, hypothyroidism, uremia, vitamin deficiency, chemotherapy-induced pain, 2', 3'-dideoxycytidine (ddC) treatment, Guillain-Barré syndrome, or Fabry's disease. Finnerup NB et al., Pain 157(8): 1599-1606(2016); National Institute of Neurological Disorders and Stroke, PeripheralNeuropathy Fact Sheet, available at ninds.nih.gov / disorders / peripheralneuropathy / detail_peripheralneuropathy.htm. In some embodiments, polyneuropathy is diabetic peripheral neuropathy. In certain embodiments, diabetic peripheral neuropathy is caused by high blood sugar levels (glucose) and / or high fat levels (eg, triglycerides) in the blood of a diabetic subject, which can cause damage to the patient's peripheral nerves.

[0060] In some embodiments, the peripheral neuropathy disclosed herein can be classified according to the partially damaged or affected nerve cells (e.g., axons, myelin sheaths, or cell bodies). In some embodiments, the peripheral neuropathy is a distal axonopathy caused by a metabolic or toxic disorder of the axons. In some embodiments, the metabolic disorder includes diabetes, renal failure, deficiency syndromes such as malnutrition and alcoholism. In some embodiments, the metabolic disorder is diabetes, and the distal axonopathy is diabetic neuropathy.

[0061] In some embodiments, peripheral neuropathy is a myelin disease caused by a primary attack on the myelin sheath or myelinated Schwann cells, resulting in acute failure of impulse conduction. The most common cause is acute inflammatory demyelinating polyneuropathy (AIDP; also known as Guillain-Barré syndrome), but other causes include chronic inflammatory demyelinating syndrome (CIDP), inherited metabolic disorders (e.g., leukodystrophy), or toxins.

[0062] In some embodiments, peripheral neuropathy is a neuropathy caused by the destruction of neurons of the peripheral nervous system (PNS). In some embodiments, the neuropathy is caused by motor neuron disease, sensory neuropathy (e.g., herpes nodularis), toxins, or autonomic dysfunction. In some embodiments, the neuropathy is caused by neurotoxins (such as the chemotherapeutic agent vincristine).

[0063] Neuropathic pain can be caused by or associated with a variety of etiologies (e.g., physical injury (e.g., trauma or repetitive stress), disease or disorder, exposure to toxic agents, or a combination thereof). In some embodiments, neuropathic pain is caused by or associated with a traumatic injury or damage, such as a nerve compression injury (e.g., nerve crush, nerve stretch, nerve entrapment, or incomplete nerve transection); spinal cord injury (e.g., half of the spinal cord); injury or damage to a peripheral nerve (e.g., motor nerves, sensory nerves, or autonomic nerves, or a combination thereof), amputation; contusion; inflammation (e.g., inflammation of the spinal cord); or surgery. In some embodiments, neuropathic pain is caused by or associated with repetitive stress, including, for example, repetitive, clumsy, and / or compulsive activities that require prolonged movement of any joint group. Without being bound by any one theory, the resulting irritation can cause inflammation and swelling of ligaments, tendons, and muscles, constricting the narrow passages through which nerves pass (e.g., ulnar neuropathy and carpal tunnel syndrome, a neuropathy caused by a restricted or compressed nerve at the elbow or wrist). In some embodiments, neuropathic pain is caused by or is associated with a disease or disorder including, for example, an ischemic event (e.g., a stroke or heart attack), multiple sclerosis, metabolic and / or endocrine diseases or disorders (e.g., diabetes, metabolic diseases, and acromegaly, a condition caused by overproduction of growth hormone and characterized by abnormal enlargement of portions of the skeleton (including joints), leading to nerve entrapment and pain), small vessel disease that results in reduced oxygen supply to peripheral nerves (leading to nerve tissue damage) (e.g., vasculitis, i.e., inflammation of blood vessels), autoimmune diseases (e.g., Sjögren's syndrome, lupus, rheumatoid arthritis, and acute inflammatory demyelinating neuropathy, also known as Guillain-Barré syndrome), renal disorders, cancer or tumors (e.g., neoplastic tumors, neuromas, paraneoplastic syndromes, and other diseases resulting from cancer treatment). In some embodiments, the neuropathic pain is caused by or is associated with an infectious agent (e.g., a tick-borne infection, varicella zoster, Epstein-Barr virus, West Nile virus, cytomegalovirus, and herpes simplex virus, acquired immunodeficiency syndrome (AIDS), or an infection caused by bacteria (such as Lyme disease, diphtheria, and leprosy), an inflammatory disorder, a peripheral nerve disorder (e.g., a neuroma), an inherited or emerging inherited disorder (e.g., Charcot-Marie-Tooth disorder includes extreme weakness and wasting of the muscles of the lower leg and foot, abnormal gait, loss of tendon reflexes, and numbness of the lower extremities), a mononeuropathy, or a polyneuropathy. In some embodiments, the neuropathic pain is caused by or is associated with an infectious agent (e.g., a tick-borne infection, varicella zoster, Epstein-Barr virus, West Nile virus, cytomegalovirus, herpes simplex virus, AIDS).In some embodiments, neuropathic pain is caused by or is associated with exposure to toxic agents, including, for example, drugs, alcohol, heavy metals (e.g., lead, arsenic, mercury), industrial agents (e.g., solvents, fumes from glue), or nitrous oxide.

[0064] The term "neuropathic pain associated with" a disease or disorder refers to neuropathic pain that accompanies, is caused by, or results from a disease or disorder (eg, those disclosed herein).

[0065] As used herein, the terms "treat," "treating," and "treatment" refer to any type of intervention or process performed on a subject, or the administration of an active agent to a subject, with the goal of reversing, alleviating, ameliorating, inhibiting, slowing down, or preventing the progression, development, severity, or recurrence of symptoms, complications, conditions, or biochemical markers associated with a disease. Treatment can be directed to a subject with a disease or a subject without the disease (e.g., for prevention).

[0066] As used herein, "administering" refers to the physical introduction of a therapeutic agent or a composition comprising a therapeutic agent into a subject using any of the various methods and delivery systems known to those skilled in the art. Preferred routes of administration for antibodies described herein include intravenous, intraperitoneal, intramuscular, subcutaneous, intraspinal, intravitreal, or other parenteral routes of administration, such as by injection or infusion. As used herein, the phrase "parenteral administration" refers to administration methods other than enteral and topical administration, typically by injection, and includes but is not limited to intravenous, intraperitoneal, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intratracheal, subcutaneous, subcutaneous, intravitreal, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, and in vivo electroporation. Alternatively, the antibodies described herein may be administered by non-parenteral routes, such as topical, epidermal, or mucosal routes of administration, such as intranasal, oral, vaginal, rectal, sublingual, or topical administration. Administration can also be performed, for example, once, multiple times, and / or over one or more extended periods of time.

[0067] As used herein, the term "therapeutically effective amount" refers to the amount of a drug, alone or in combination with another therapeutic agent, that effectively "treats" a subject's disease or disorder or reduces the risk, potential, probability, or incidence of a disease or disorder (e.g., neuropathic pain). A "therapeutically effective amount" includes the amount of a drug or therapeutic agent that provides some improvement or benefit to a subject who is at risk of or has a disease or disorder (e.g., neuropathic pain disclosed herein). Therefore, a "therapeutically effective" amount refers to an amount that reduces the risk, potential, probability, or incidence of a disease or disorder, or alleviates, mitigates, and / or reduces at least one indicator (e.g., neuropathic pain), and / or mitigates at least one clinical symptom of a disease or disorder.

[0068] As used herein, the term "subject" includes any human or non-human animal. The term "non-human animal" includes all vertebrates, e.g., mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc.

[0069] The term "sequence similarity family 19 member A5" or "FAM19A5" refers to a protein that belongs to the TAFA family of five highly homologous proteins (also known as the FAM19 family) and is primarily expressed in the brain and spinal cord. FAM19A5 is also known as TAFA5 or chemokine-like protein TAFA-5.

[0070] In humans, the gene encoding FAM19A5 is located on chromosome 22. There are multiple human FAM19A5 (UniProt: Q7Z5A7) isoforms, which are thought to be produced by alternative splicing: isoform 1 (UniProt: Q7Z5A7-1), which consists of 132 amino acids; isoform 2 (UniProt: Q7Z5A7-2), which consists of 125 amino acids; and isoform 3 (UniProt: Q7Z5A7-3), which consists 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 with one transmembrane region. Isoform 2, reported as a secretory protein (soluble) in Tang TY et al., Genomics 83 (4): 727-34 (2004), contains a signal peptide at amino acid positions 1-25. Isoform 1 is thought to be a membrane protein. Below are the amino acid sequences of the three known human FAM19A5 isoforms.

[0071] (I) Isoform 1 (UniProt: Q7Z5A7-1, transmembrane protein): This isoform has been selected as the canonical sequence.

[0072] MAPSPRTGSR QDATALPSMS STFWAFMILA SLLIAYCSQL AAGTCEIVTL DRDSSQPRRTIARQTARCAC RKGQIAGTTR ARPACVDARI IKTKQWCDML PCLEGEGCDL LINRSGWTCT QPGGRIKTTTVS(SEQ ID NO:1)

[0073] (II) Isoform 2 (UniProt: Q7Z5A7-2, soluble protein):

[0074] MQLLKALWAL AGAALCCFLV LVIHAQFLKE GQLAAGTCEI VTLDRDSSQP RRTIARQTARCACRKGQIAG TTRARPACVD ARIIKTKQWC DMLPCLEGEG CDLLINRSGW TCTQPGGRIK TTTVS(SEQID NO:2)

[0075] (III) Isoform 3 (UniProt: Q7Z5A7-3):

[0076] MYHHREWPAR IIKTKQWCDM LPCLEGEGCD LLINRSGWTC TQPGGRIKTT TVS(SEQ ID NO:3)

[0077] The term "FAM19A5" includes any variant or isoform of FAM19A5 naturally expressed by cells. Therefore, the antibodies described herein can cross-react with different isoforms in the same species (e.g., different isoforms of human FAM19A5), or cross-react with FAM19A5 (e.g., mouse FAM19A5) from species other than humans. Alternatively, the antibody can be specific to human FAM19A5 and cannot show any cross-reactivity with other species. FAM19A5 or any variant and isoform thereof can be separated or recombinantly produced from cells or tissues that naturally express them. The polynucleotide encoding human FAM19A5 has GenBank accession number BC039396 and the following sequence:

[0078] Table 1 Polynucleotide sequence of human FAM19A5

[0079]

[0080] The term "antagonist to FAM19A5 protein" refers to all antagonists that inhibit the expression of FAM19A5 protein. Such antagonists can be peptides, nucleic acids or compounds. More specifically, the antagonist can be an antisense oligonucleotide, siRNA, shRNA, miRNA, dsRNA, aptamer, PNA (peptide nucleic acid) targeting FAM19A5 or a vector comprising them. In some embodiments, the antagonist can be an antibody or an antigen binding portion thereof that specifically binds to the FAM19A5 protein.

[0081] The terms "antibody" and "antibodies" are terms of the art and can be used interchangeably herein and refer to molecules with an antigen binding site that specifically binds to an antigen. As used herein, the term includes a complete antibody and any antigen binding fragment (i.e., "antigen binding portion") or a single chain thereof. In some embodiments, an "antibody" refers to a glycoprotein or an antigen binding portion thereof comprising at least two heavy chains (H) and two light chains (L) interconnected by a disulfide bond. In other embodiments, an "antibody" refers to a single-chain antibody comprising a single variable domain, such as 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.

[0082] 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. 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.

[0083] The term "Kabat numbering" and similar terms are well-known in the art, and refer to a system for numbering the amino acid residues in the heavy chain and light chain variable region of an antibody or its antigen-binding portion thereof. In some aspects, the CDR of an antibody can be determined according to the Kabat numbering system (see, e.g., Kabat EA & Wu TT (1971) Ann NY Acad Sci 190: 382-391 and Kabat EA et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242). Using the Kabat numbering system, the CDR in an antibody heavy chain molecule is usually present in amino acid positions 31 to 35, which optionally can include one or two additional amino acids after 35 (referred to as 35A and 35B in the Kabat numbering scheme) (CDR1), amino acid positions 50 to 65 (CDR2) and amino acid positions 95 to 102 (CDR3). Using the Kabat numbering system, the CDRs within an antibody light chain molecule are typically present at amino acid positions 24 to 34 (CDR1), amino acid positions 50 to 56 (CDR2), and amino acid positions 89 to 97 (CDR3). In a specific embodiment, the CDRs of the antibodies described herein are determined according to the Kabat numbering scheme.

[0084] The phrase "amino acid position numbering as in Kabat", "Kabat position" and grammatical variants thereof refer to the numbering system for the heavy chain variable domain or light chain variable domain of the antibody compilation in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991). Using this numbering system, the actual linear amino acid sequence may correspond to a shortening or insertion of the FW or CDR of the variable domain containing fewer or additional amino acids. 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 heavy chain FW residue 82 (e.g., residues 82a, 82b, and 82c, etc. according to Kabat). See Table 1B.

[0085] Table 1B

[0086]

[0087]

[0088] For a given antibody, the Kabat numbering of the residues can be determined by comparing the homology regions of the antibody's sequence with the sequence of the "standard" Kabat numbering. Chothia refers to the position of the structural loop (Chothia and Lesk, J. Mol. Biol. 196: 901-917 (1987)). When numbering using the Kabat numbering convention, the end of the Chothia CDR-H1 loop varies between H32 and H34, depending on the length of the loop (this is because the Kabat numbering scheme places the insertion point 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 region represents a compromise between the Kabat CDR and the Chothia structural loops and is used by Oxford Molecular's AbM antibody modeling software.

[0089] IMGT (ImMunoGeneTics) also provides a numbering system for immunoglobulin variable regions (including CDRs). See, e.g., Lefranc, MP et al., Dev. Comp. Immunol. 27:55-77 (2003), which is incorporated herein by reference. The IMGT numbering system is based on the alignment of more than 5,000 sequences, structural data, and characterization of hypervariable loops, and allows easy comparison of variable regions and CDR regions of all species. According to the IMGT numbering scheme, VH-CDR1 is located at positions 26 to 35, VH-CDR2 is located at positions 51 to 57, VH-CDR3 is located at positions 93 to 102, VL-CDR1 is located at positions 27 to 32, VL-CDR2 is located at positions 50 to 52, and VL-CDR3 is located at positions 89 to 97.

[0090] 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. USA 63 (1): 78-85, which describes the amino acid sequence of the first human IgG1 sequenced myeloma protein EU. The EU index of Edelman et al. is also described in Kabat et al., 1991, Sequences of Proteins of Immunological Interest, 5th Ed., United States Public Health Service, National Institutes of Health, Bethesda. Thus, the phrases "EU index as described in Kabat" or "EU index of Kabat" and "position according to the EU index as described in Kabat" and grammatical variants thereof refer to the residue numbering system for the human IgG1 EU antibody based on Edelman et al., as described in Kabat 1991.

[0091] The numbering system for variable domains (heavy and light chains) and light chain constant region amino acid sequences is described in Kabat 1991.

[0092] The antibody can be any type of immunoglobulin molecule (e.g., IgG, IgE, IgM, IgD, IgA or IgY), any species (e.g., IgD, IgG2, IgG3, IgG4, IgA1 or IgA2) or any subclass (e.g., IgG1, IgG2, IgG3 and IgG4 of humans, and IgG1, IgG2a, IgG2b and IgG3 of mice). Immunoglobulins, such as IgG1, exist in several allotypes, which differ from each other in a few amino acids at most. The antibodies disclosed herein can be from any known isotype, class, subclass or allotype. In certain embodiments, the antibodies described herein are IgG1, IgG2, IgG3 or IgG4 subclasses or any hybrids thereof. In certain embodiments, the antibody is a human IgG1 subclass, a human IgG2 subclass or a human IgG4 subclass.

[0093] "Antibodies" include, for example, naturally occurring and non-naturally occurring antibodies; monoclonal antibodies 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; heterotypic conjugate antibodies; monovalent antibodies; single chain antibodies; camelid antibodies; affybodies; 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 binding to an antigen. Harmen MM and Haard HJ Appl Microbiol Biotechnol. 77(1):13-22 (2007)).

[0094] As used herein, the term "antigen binding portion" and "antigen binding fragment" of an antibody are interchangeable and refer to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., people FAM19A5). Such a "fragment" may have a length of, for example, between about 8 and about 1500 amino acids, a length of, for example, between about 8 and about 745 amino acids, a length of, for example, between about 8 and about 300 amino acids, such as about 8 to about 200 amino acids, or about 10 to about 50 or 100 amino acids. It has been shown that the antigen binding function of an antibody can be achieved by a fragment of a full-length antibody. Examples of binding fragments encompassed by the term "antigen-binding portion" of an antibody, e.g., an anti-FAM19A5 antibody 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) an Fv fragment consisting of the VL and VH domains of a single arm of an antibody, and a disulfide-linked Fv (SDFv); (v) a dAb fragment consisting of a VH domain (Ward et al., (1989) Nature 341:544-546); and (vi) an isolated complementarity determining region (CDR) or (vii) a combination of two or more isolated CDRs, which may optionally be linked by a synthetic linker. In addition, although the two domains VL and VH of the Fv fragment are encoded by different genes, they can be connected using recombinant methods through synthetic linkers, enabling them to be formed into a single protein chain, in which the VL and VH regions are paired to form a monovalent molecule (referred to as single-chain Fv (ScFv); see, e.g., 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 intended to be included in the term "antigen-binding portion" of an antibody. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and the usefulness of the fragments is screened in the same manner as intact antibodies. The antigen-binding portion can be produced by recombinant DNA technology, or by enzymatic or chemical cleavage of intact immunoglobulins.

[0095] As used herein, the terms "variable region" and "variable domain" are used interchangeably and are common in the art. A variable region generally refers to a portion of an antibody, typically a portion of a light chain or a heavy chain, typically the amino terminal of about 110 to 120 amino acids in a mature heavy chain and about 90 to 115 amino acids in a mature light chain, which differ 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 those regions known as complementary determining regions (CDRs), while the more highly conserved regions in the variable domains are known as framework regions (FRs).

[0096] Without wishing to be bound by any particular mechanism or theory, the CDRs of light and heavy chains are considered to be primarily responsible for the interaction and specificity of the antibody with the antigen. In certain embodiments, the variable region is a human variable region. In certain embodiments, the variable region includes rodent or mouse CDRs and human framework regions (FRs). In specific embodiments, the variable region is a primate (e.g., non-human primate) variable region. In certain embodiments, the variable region includes rodent or mouse CDRs and primate (e.g., non-human primate) framework regions (FRs).

[0097] As used herein, the term "heavy chain" (HC) when used in reference to antibodies can refer to any of the different types, such as alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), based on the amino acid sequence of the constant domain, which give rise to IgA, IgD, IgE, IgG, and IgM class antibodies, respectively, including subclasses of IgG, such as IgG1, IgG2, IgG3, and IgG4.

[0098] As used herein, the term "light chain" (LC) when referring to an antibody can refer to any of the different types, such as kappa (κ) or lambda (λ) based on the amino acid sequence of the constant domain. Light chain amino acid sequences are well known in the art. In a specific embodiment, the light chain is a human light chain.

[0099] The terms "VL" and "VL domain" are used interchangeably to refer to the light chain variable region of an antibody.

[0100] The terms "VH" and "VH domain" are used interchangeably to refer to the heavy chain variable region of an antibody.

[0101] As used herein, the terms "constant region" and "constant domain" are interchangeable and have their general 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 the binding of the antibody to an antigen, but may exhibit various effector functions, such as interactions with Fc receptors. The constant region of an immunoglobulin molecule generally has a more conserved amino acid sequence relative to an immunoglobulin variable domain.

[0102] "Fc region" (fragment crystallizable region) or "Fc domain" or "Fc" refers to the C-terminal region of the heavy chain of an antibody, which mediates the binding of the immunoglobulin to host tissues or factors, including binding to Fc receptors located on various cells of the immune system (e.g., effector cells) or binding 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 contains two identical protein fragments of the second (CH2) and third (CH3) constant domains derived from the two heavy chains of the antibody; IgM and IgE Fc regions contain 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 human IgG heavy chain Fc region 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 according to the EU index as 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., it extends from about amino acid 341 of IgG to about amino acid 447. As used herein, the Fc region may be a native sequence Fc, including any allotype variants, or a variant Fc (e.g., a non-naturally occurring Fc). Fc may also refer to this region in the context of an isolated or Fc-containing protein polypeptide (such as a "binding protein comprising an Fc region", also referred to as an "Fc fusion protein" (e.g., an antibody or immunoadhesin)).

[0103] A "native sequence Fc region" or "native sequence Fc" comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature. 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 allotypes of Fcs (see, e.g., Jefferis et al., (2009) mAbs 1:1; Vidarsson G et al. Front Immunol. 5:520 (published online on October 20, 2014)).

[0104] "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 FcγR family, including allelic variants and alternatively spliced ​​forms of these receptors. The FcγR family consists of three activating (FcγRI, FcγRIII and FcγRIV in mice; FcγRIA, FcγRIIA and FcγRIIIA in humans) and one inhibitory (FcγRIIB) receptor. Human IgG1 binds to most human Fc receptors and triggers the strongest Fc effector function. In terms of the type of activated Fc receptor it binds, it is considered to be equivalent to mouse IgG2a. In contrast, human IgG4 triggers the least Fc effector function. Vidarsson G et al., Front Immunol.5:520 (published online on October 20, 2014).

[0105] The constant region can be operated by, for example, recombinant technology to eliminate one or more effector functions. "Effector function" refers to the interaction of an 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 that the Fc region be combined with a binding domain (e.g., an antibody variable domain). Therefore, the term "constant region without Fc function" includes a constant region with reduced or no one or more effector functions mediated by the Fc region.

[0106] 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., such as 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, so-called glycosylated antibodies can be produced by removing sugars attached to specific residues in the Fc region to reduce the effector function of the antibody while retaining other valuable properties of the Fc region (e.g., extended half-life and heterodimerization). Glycosylated antibodies can be produced 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 by expressing antibodies in cells (e.g., bacterial host cells) that cannot glycosylate proteins. See, for example, U.S. Patent Publication No. 20120100140. Another method is to use an Fc region from an IgG subclass with reduced effector function, for example, IgG2 and IgG4 antibodies are characterized by having a lower level of Fc effector function than IgG1 and IgG3. The residues closest to the hinge region in the CH2 domain of the Fc portion are responsible for the effector function of the antibody because it contains largely 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 October 20, 2014). Therefore, antibodies with reduced or no Fc effector function can be prepared by: generating, for example, a chimeric Fc region comprising a CH2 domain of an IgG antibody from an IgG4 isotype and a CH3 domain of an IgG antibody from an IgG1 isotype; or a chimeric Fc region comprising a hinge region from IgG2 and a CH2 region from IgG4 (see, e.g., Lau C et al., J. Immunol. 191: 4769-4777 (2013)); or an Fc region with a mutation that results in an altered Fc effector function (e.g., reduced or no Fc function). Such Fc regions with mutations are known in the art. See, e.g., U.S. Patent Publication No. 20120100140 and the U.S. and PCT applications cited therein and An et al., mAbs 1:6, 572-579 (2009); the disclosures of which are incorporated herein by reference in their entirety.

[0107] "Hinge", "hinge domain" or "hinge region" or "antibody hinge region" refers to the domain of the heavy chain constant region connecting the CH1 domain to the CH2 domain, and includes the upper, middle and lower parts of the hinge (Roux et al., J. Immunol. 1998 161: 4083). The hinge provides varying degrees of flexibility between the binding region and the effector region of the antibody, and also provides a site for intermolecular disulfide bonding between the two heavy chain constant regions. As used herein, for all IgG isotypes, the hinge starts at Glu216 and ends at Gly 237 (Roux et al., 1998 J Immunol 161: 4083). The sequences of wild-type IgG1, IgG2, IgG3 and IgG4 hinges are known in the art. See, e.g., Kabat EA et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242; Vidarsson G et al., Front Immunol. 5:520 (published online Oct. 20, 2014).

[0108] The term "CH1 domain" refers to the heavy chain constant region that connects the variable domain to the hinge in the heavy chain constant domain. As used herein, the CH1 domain starts at A118 and ends at V215. The term "CH1 domain" includes wild-type CH1 domains and naturally occurring variants thereof (e.g., allotypes). The CH1 domain sequences of IgG1, IgG2, IgG3, and IgG4 (including wild-type and allotypes) are known in the art. See, e.g., Kabat EA et al., (1991) supra and Vidarsson G et al., Front Immunol.5:520 (published online on October 20, 2014). Exemplary CH1 domains include CH1 domains having mutations that change the biological activity (e.g., half-life) of antibodies, such as described in U.S. Patent Publication No. 20120100140 and the U.S. Patents and Publications cited therein and PCT Publications.

[0109] The term "CH2 domain" refers to the heavy chain constant region that connects the hinge to the CH3 domain in the heavy chain constant domain. As used herein, the CH2 domain starts at P238 and ends at K340. The term "CH2 domain" includes wild-type CH2 domains and naturally occurring variants thereof (e.g., allotypes). The CH2 domain sequences of IgG1, IgG2, IgG3, and IgG4 (including wild-type and allotypes) are known in the art. See, e.g., Kabat EA et al., (1991) supra and Vidarsson G et al., 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., half-life and / or reduced Fc effector functions), such as described in U.S. Patent Publication No. 20120100140 and the U.S. Patents and Publications cited therein and PCT Publications.

[0110] The term "CH3 domain" refers to the heavy chain constant region that is the C-terminal end of the CH2 domain in the heavy chain constant domain. As used herein, the CH3 domain starts at G341 and ends at K447. The term "CH3 domain" includes wild-type CH3 domains and naturally occurring variants thereof (e.g., allotypes). The CH3 domain sequences of IgG1, IgG2, IgG3, and IgG4 (including wild-type and allotypes) are known in the art. See, e.g., Kabat EA et al., (1991) supra and Vidarsson G et al., Front Immunol.5:520 (published online on October 20, 2014). Exemplary CH3 domains include CH3 domains with mutations that change the biological activity (e.g., half-life) of antibodies, such as described in U.S. Patent Publication No. 20120100140 and the U.S. Patents and Publications cited therein and PCT Publications.

[0111] As used herein, "isotype" refers to the antibody class encoded by the heavy chain constant region genes (e.g., IgGl, IgG2, IgG3, IgG4, IgM, IgAl, IgA2, IgD, and IgE antibodies).

[0112] "Allotype" refers to naturally occurring variants within a particular isotype group that differ in a few amino acids (see, e.g., Jefferis et al., (2009) mAbs 1:1). The antibodies described herein can be of any allotype. Allotypes of IgG1, IgG2, IgG3, and IgG4 are known in the art. See, e.g., Kabat EA et al., (1991) supra; Vidarsson G et al., Front Immunol. 5:520 (published online Oct. 20, 2014); and Lefranc MP, mAbs 1:4, 1-7 (2009).

[0113] The phrases "an antibody that recognizes an antigen" and "an antibody specific for an antigen" are used interchangeably herein with the term "an antibody that specifically binds to an antigen."

[0114] As used herein, "isolated antibody" means an antibody that is 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 an epitope of FAM19A5 may have cross-reactivity with other FAM19A5 proteins from different species.

[0115] "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 represented by a dissociation constant (KD). Affinity can be measured and / or expressed in a variety of ways known in the art, including, but not limited to, an equilibrium dissociation constant (KD) and an equilibrium association constant (KA). KD is calculated by the quotient of koff / kon and is expressed as a molar concentration (M), while KA is calculated by the quotient of kon / koff. kon refers to, for example, the association rate constant of an antibody with an antigen, and koff refers to, for example, the dissociation of an antibody from an antigen. kon and koff can be determined by techniques known to those of ordinary skill in the art, such as immunoassays (e.g., enzyme-linked immunosorbent assay (ELISA)), or kinetic exclusion assay (KinExA).

[0116] As used herein, the terms "specifically binds," "specifically recognizes," "specifically binds," "selectively binds," and "selectively binds" 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), as such binding is understood by those skilled in the art. For example, a molecule that specifically binds to an antigen can typically be detected, for example, by immunoassays, The molecule specifically binds to the antigen with a KA that is at least 2 log, 2.5 log, 3 log, 4 log or greater than the KA when the molecule is bound to another antigen.

[0117] Antibodies are usually specifically bound to their cognate antigens with a high affinity reflected by a dissociation constant (KD) of 10-5 to 10-11 M or less. Any KD greater than about 10-4 M is generally considered to be non-specific binding. As used herein, an antibody that "specifically binds" to an antigen refers to an antibody that binds to an antigen and substantially the same antigen with high affinity, which means that when determined by, for example, an immunoassay (e.g., ELISA) or a surface plasmon resonance (SPR) technique in a BIACORE 2000 instrument using a predetermined antigen, it has 10-7 M or less, preferably 10-8 M or less, even more preferably 10-9 M or less, and most preferably between 10-8 M and 10-10 M or less KD, but not bound to an irrelevant antigen with high affinity.

[0118] 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.

[0119] As used herein, "epitope" is a term in the art, and refers to the local region of the antigen that an antibody can specifically bind. An epitope can be, for example, the continuous amino acids (linear or continuous epitopes) of a polypeptide, or an epitope can be, for example, two or more discontinuous regions (conformation, nonlinearity, discontinuous or discontinuous epitopes) from one or more polypeptides. The epitope formed by continuous amino acids is usually, but not always, retained when exposed to a denaturing solvent, and the epitope formed by the tertiary folding is usually lost when treated with a denaturing solvent. An epitope generally includes at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 20 amino acids of a unique spatial conformation. The method for determining the epitope combined by a given antibody (i.e., epitope mapping) is well known in the art, and includes, for example, immunoblotting and immunoprecipitation assays, wherein the test is from (for example, from FMAM19A5) overlapping or adjacent peptides with the reactivity of a given antibody (for example, anti-FAM19A5 antibodies). Methods for determining the spatial conformation of epitopes include techniques in the art and those described herein, e.g., x-ray crystallography, two-dimensional nuclear magnetic resonance, and HDX-MS (see, e.g., Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, GE Morris, Ed. (1996)).

[0120] In certain embodiments, the epitope to which the antibody binds can be determined by, for example, NMR spectroscopy, X-ray diffraction crystallography, ELISA assays, hydrogen / deuterium exchange in combination with 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, any method known in the art can be used to complete crystallization (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, distributed by Molecular Simulations, Inc.; see, e.g., Meth Enzymol (1985) volumes 114 & 115, eds Wyckoff HW et al.; US 2004 / 0014194), and BUSTER (Bricogne G (1993) Acta Crystallogr D Biol Crystallogr 49 (Pt 1): 37-60; Bricogne G (1997) Meth Enzymol 276A: 361-423, ed Carter CW; Roversi P et al., (2000) Acta Crystallogr D Biol Crystallogr 56 (Pt 10): 1316-1323). Mutagenesis mapping studies can be performed using any method known to those skilled in the art. See, for example, Champe M et al., (1995) J Biol Chem 270: 1388-1394 and Cunningham BC & Wells JA (1989) Science 244: 1081-1085 for a description of mutagenesis techniques including alanine scanning mutagenesis.

[0121] The term "epitope mapping" refers to the process of identifying molecular determinants for antibody-antigen recognition.

[0122] The term "binding to the same epitope" referring to two or more antibodies refers to the same segment of amino acid residues as determined by a given method. The technology used to determine whether an antibody is bound to the same epitope on FAM19A5 as an antibody as described herein includes, for example, epitope mapping methods, such as X-ray analysis (which provides atomic resolution of epitopes) and hydrogen / deuterium exchange mass spectrometry (HDX-MS) of crystals to antigen: antibody complexes. Other methods monitor the binding of antibodies to variants of mutations of antigen fragments or antigens, wherein the lack of binding caused by the modification of amino acid residues in the antigen sequence is generally considered to be an indication of epitope components. In addition, the computational combination method for epitope mapping can also be used. These methods rely on the ability of the antibody of interest to affinity separate specific short peptides from the combined phage display peptide library. Antibodies with identical VH and VL or identical CDR1, 2 and 3 sequences are expected to bind to the same epitope.

[0123] An antibody that "competes with another antibody for binding to a target" refers to an antibody that inhibits (partially or completely) another antibody from binding to a target. Whether two antibodies compete with each other for binding to a target, i.e., whether and to what extent one antibody inhibits another antibody from binding to a target, can be determined using known competition experiments. In certain embodiments, an antibody competes with another antibody and inhibits another antibody from binding to a target by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%. Depending on which antibody is a "blocking antibody" (i.e., a cold antibody that is first incubated with the target), the level of inhibition or competition may be different. Competition assays can be performed as described, for example, in Ed Harlow and David Lane, Cold Spring Harb Protoc; 2006; doi: 10.1101 / pdb.prot4277 or in Chapter 11 of "Using Antibodies" by Ed Harlow and David Lane, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA 1999. Competing antibodies bind to the same epitope, overlapping epitopes, or adjacent epitopes (e.g., as demonstrated by steric hindrance).

[0124] Other competitive binding assays include: solid phase direct or indirect radioimmunoassay (RIA), solid phase direct or indirect enzyme immunoassay (EIA), sandwich competition assay (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 labeling assay, solid phase direct labeling sandwich assay (see Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Press (1988)); solid phase direct labeling RIA using 1-125 label (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)).

[0125] "Bispecific" or "bifunctional antibodies" are artificial hybrid antibodies with two different heavy / 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. See, e.g., Songsivilai & Lachmann, Clin. Exp. Immunol. 79:315-321 (1990); Kostelny et al., J. Immunol. 148, 1547-1553 (1992).

[0126] As used herein, the term "monoclonal antibody" refers to an antibody that displays a single binding specificity and affinity for a specific epitope, or a composition of antibodies in which all antibodies display a single binding specificity and affinity for a specific epitope. Therefore, 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 some embodiments, human monoclonal antibodies are produced by hybridomas, which include B cells fused to immortalized cells obtained from transgenic non-human animals (e.g., transgenic mice), which have a genome comprising human heavy chain transgenes and light chain transgenes.

[0127] As used herein, the term "recombinant human antibody" includes all human antibodies prepared, expressed, produced or separated by recombinant means, such as (a) antibodies isolated from transgenic or transchromosomal animals (e.g., mice) to human immunoglobulin genes or hybridomas prepared therefrom, (b) antibodies isolated from host cells converted to express antibodies, such as antibodies isolated from transfectomas, (c) antibodies isolated from recombinant combinatorial human antibody libraries, and (d) antibodies prepared, expressed, produced or separated by any other means involving splicing of human immunoglobulin gene sequences to DNA sequences. Such recombinant human antibodies include variable regions and constant regions, which utilize specific human germline immunoglobulin sequences encoded by germline genes, but include subsequent rearrangements and mutations that occur, for example, during antibody maturation. As known in the art (see, e.g., Lonberg (2005) Nature Biotech. 23 (9): 1117-1125), variable regions contain antigen-binding domains encoded by various genes, which are rearranged to form antibodies specific to exogenous antigens. In addition to rearrangement, the variable region can be further modified by multiple single amino acid changes (known as somatic mutations or hypermutations) to increase the affinity of the antibody for foreign antigens. The constant region will further change in response to the antigen (i.e., isotype switching). Thus, the rearranged and somatically mutated nucleic acid molecules encoding the light and heavy chain immunoglobulin polypeptides that respond to the antigen cannot have sequence identity with the original nucleic acid molecules, but will be substantially identical or similar (i.e., have at least 80% identity).

[0128] "HuMAb" refers to an antibody with a variable region, wherein the framework region and the CDR region are all from human germline immunoglobulin sequences. In addition, if the antibody contains a constant region, the constant region is also derived from human germline immunoglobulin sequences. 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, as used herein, the term "human antibody" 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 terms "human" antibody and "fully human" antibody are used synonymously.

[0129] "Humanized" antibodies refer to antibodies in which some, most or all of the amino acids outside the CDR domains of non-human antibodies are replaced by the corresponding amino acids derived from human immunoglobulins. In some embodiments of the humanized form of the antibody, some, most or all of the amino acids outside the CDR domains have been replaced by amino acids from human immunoglobulins, while some, most or all of the amino acids in one or more CDR regions are unchanged. Small additions, deletions, insertions, substitutions or modifications of amino acids are allowed as long as they do not destroy the ability of the antibody to bind to a specific antigen. "Humanized" antibodies retain antigenic specificity similar to the original antibody.

[0130] "Chimeric antibodies" refer to antibodies in which the variable regions are derived from one species and the constant regions are derived from another species, such as antibodies in which the variable regions are derived from a mouse antibody and the constant regions are derived from a human antibody.

[0131] As used herein, the term "cross-reaction" refers to the ability of antibodies described herein to bind to FAM19A5 from different species. For example, antibodies described herein that bind to human FAM19A5 may also bind to another species of FAM19A5 (e.g., mouse FAM19A5). As used herein, cross-reactivity can be measured by detecting specific reactivity with purified antigens in binding assays (e.g., SPR, ELISA), or by binding to cells that physiologically express FAM19A5, or otherwise interacting with the cell function. Methods for determining cross-reactivity include standard binding assays as described herein, for example, by using Biacore TM Biacore 2000 SPR instrument (Biacore AB, Uppsala, Sweden) TM Surface plasmon resonance (SPR) analysis, or flow cytometry techniques.

[0132] As used herein, the term "naturally occurring" as applied to an object means that the object can be found in nature. For example, a polypeptide or polynucleotide sequence present in an organism (including a virus) is naturally occurring, as can be isolated from a source in nature and has not been intentionally modified by humans in the laboratory.

[0133] "Polypeptide" refers to a chain of at least two consecutively linked amino acid residues, with no upper limit to 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 contain one or more polypeptides.

[0134] As used herein, the term "nucleic acid molecule" is intended to include DNA molecules and RNA molecules. The nucleic acid molecule may be single-stranded or double-stranded, and may be a cDNA.

[0135] As used herein, the term "vector" means 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 in which an additional DNA segment can be connected. Another type of vector is a viral vector, in which an additional DNA fragment can be connected to the viral genome. Some vectors can replicate autonomously in the host cell into which they are introduced (for example, bacterial vectors and episomal mammalian vectors with bacterial replication origins). Other vectors (for example, non-episomal mammalian vectors) can be integrated into the genome of the host cell after being introduced into the host cell, and thus replicated together with the host genome. In addition, some vectors can guide the expression of the gene effectively connected thereto. Such a vector is referred to herein as a "recombinant expression vector" (or simply referred to as "expression vector"). Generally, an expression vector useful in recombinant DNA technology is usually in the form of a plasmid. In this specification, "plasmid" and "vector" can be used interchangeably because plasmid is the most commonly used vector form. However, other forms of expression vectors are also included, such as viral vectors (for example, replication-defective retroviruses, adenoviruses and adeno-associated viruses), which play equivalent functions.

[0136] As used herein, the term "recombinant host cell" (or simply "host cell") means a cell that contains a nucleic acid that is not naturally present in the cell, and may be a cell into which a recombinant expression vector has been introduced. It should be understood that such terms refer not only to a particular subject cell, but also to the progeny of such a cell. Because mutations or environmental influences may cause certain modifications to occur in subsequent generations, such progeny may not in fact be identical to the parent cell, but are still included within the scope of the term "host cell" as used herein.

[0137] As used herein, the term "linked" refers to the association of two or more molecules. The linking may be covalent or non-covalent. The linking may also be genetic (i.e., recombinant fusion). Such linking may be achieved using a variety of art-recognized techniques, such as chemical conjugation and recombinant protein production.

[0138] II. Methods for treating neuropathic pain

[0139] Disclosed herein is a method for treating neuropathic pain in a subject in need thereof, comprising administering to the subject an antagonist (e.g., a separated monoclonal antibody or an antigen binding fragment thereof) that specifically binds to FAM19A5. In some embodiments, neuropathic pain is central neuropathic pain, that is, pain caused by damage or impairment (e.g., brain injury and spinal cord injury) affecting any level of CNS including the central somatosensory nervous system, or pain associated with disease or disorder such as stroke, multiple sclerosis or dorsolateral medullary infarction or caused by it. In some embodiments, central neuropathic pain can be spontaneous or stimulus-induced. In some embodiments, central neuropathic pain can include dynamic mechanical allodynia and cold allodynia. Symptoms of central neuropathic pain include, for example, common sensations such as burning, tingling, shooting, squeezing, cold pain, paresthesia and insensitivity (e.g., tingling, numbness, cold and compression). The distribution range of central neuropathic pain includes from small area to large area, for example, in the periorbital area, or covering half of the body in stroke or covering the lower body in spinal cord injury, or involving one side of the face and the opposite side of the body or limbs. Central neuropathic pain caused by spinal cord injury includes "plane" pain and "below the plane" pain, "plane" pain is the pain perceived in segmental mode at the injury plane, and "below the plane" pain is the pain felt below the injury plane. In certain embodiments, the method slows down, reverses, alleviates, improves, inhibits or slows down or prevents central neuropathic pain, symptoms associated with pain, potential causes of pain or their combination.

[0140] In some embodiments, neuropathic pain is peripheral neuropathic pain, a pain caused by damage or impairment (e.g., damage to motor nerves, sensory nerves, autonomic nerves, or a combination thereof) at any level affecting the peripheral nervous system, or pain caused by a disease or disorder or pain associated with a disease or disorder. Damage or impairment of motor nerves is associated with symptoms such as muscle weakness (e.g., muscle weakness in the back, legs, buttocks, or face), painful cramps and fasciculations (uncontrolled muscle twitches visible under the skin), muscle atrophy (severe contraction of muscle size), and reduced reflexes. Damage or impairment of sensory nerve damage leads to a variety of symptoms, including pain and oversensitivity of pain receptors in the skin, leading to allodynia (e.g., severe pain caused by a normally painless stimulus).

[0141] In some embodiments, the method treats one or more types of neuropathic pain, comprising administering a FAM19A5 antagonist (e.g., an anti-FAM19A5 antibody or antigen-binding fragment thereof) to a subject in need thereof. In some embodiments, the neuropathic pain that can be treated with the methods disclosed herein is neuralgia, including but not limited to trigeminal neuralgia (TN) (e.g., pain in the trigeminal nerve region of the face or mouth), atypical trigeminal neuralgia (ATN), occipital neuralgia, postherpetic neuralgia (e.g., pain unilaterally distributed in one or more spinal dermatomes or trigeminal ophthalmic branches), peripheral nerve injury pain (e.g., pain in the innervation area of ​​the diseased nerve, usually outside of trauma, surgery or compression), glossopharyngeal neuralgia (e.g., irritation of the ninth cranial nerve that causes extreme pain in the throat, tongue and behind the ear), sciatica, low back pain and atypical facial pain. In some embodiments, the neuralgia is caused by or is associated with chemical irritation, inflammation, trauma (including surgery), nerve compression or infection caused by, for example, a nearby structure (e.g., a tumor). In some embodiments, neuropathic pain is a deafferentation pain syndrome, which includes but is not limited to damage to the brain or spinal cord, post-stroke pain, phantom limb pain, paraplegia, brachial plexus avulsion, lumbar radiculopathy. In some embodiments, neuropathic pain is complex regional pain syndrome (CRPS), including CRPS1 and CRPS2. CRPS includes but is not limited to, in some embodiments, symptoms associated with CRPS may include severe pain, changes in nails, bones and skin; and increased sensitivity of the affected limb to touch. In some embodiments, neuropathic pain is neuropathy (e.g., central or peripheral). Non-limiting examples of neuropathic pain include, for example, mononeuropathic pain (mononeuropathy) and polyneuropathic pain (polyneuropathy).

[0142] In some embodiments, the neuropathic pain is caused by or is associated with a physical injury, including, for example, (1) traumatic injury or damage, including nerve compression (e.g., nerve crush, nerve stretch, nerve entrapment, or incomplete nerve transection); (2) spinal cord injury (e.g., hemisection of the spinal cord); (3) injury or damage to peripheral nerves (e.g., motor nerves, sensory nerves, or autonomic nerves, or a combination thereof), (4) amputation; contusion; inflammation (e.g., inflammation of the spinal cord); or surgery; and (5) repetitive stress, including, for example, repetitive, clumsy, and / or compulsive activities requiring prolonged movement of any joint group (e.g., ulnar neuropathy and carpal tunnel syndrome). In some embodiments, the method treats neuropathic pain caused by or associated with exposure to a toxic agent.

[0143] In some embodiments, the neuropathic pain is caused by or is associated with one or more diseases or disorders, including, for example, (1) ischemic events (e.g., stroke or heart attack), (2) multiple sclerosis, (3) metabolic and / or endocrine diseases or disorders (e.g., diabetes, metabolic diseases, and acromegaly, a condition caused by excessive production of growth hormone and characterized by abnormal enlargement of skeletal parts including joints, leading to nerve entrapment and pain), (4) small vessel disease, which results in reduced oxygen supply to peripheral nerves, resulting in damage to nerve tissue (e.g., vasculitis, i.e., inflammation of blood vessels), (5) autoimmune diseases (e.g., Sjögren's syndrome, lupus, rheumatoid arthritis, and acute inflammatory demyelinating neuropathy, also known as Guillain-Barré syndrome), (6) renal disorders, (7) cancer or tumor (e.g., neoplastic tumors, neuromas, paraneoplastic syndromes, and toxicity from chemotherapeutic agents and radiation in cancer treatment), (8) infection (e.g., infection caused by viruses such as varicella zoster (shingles), Epstein-Barr virus, West Nile virus, cytomegalovirus, and herpes simplex virus, AIDS, or infection caused by bacteria such as Lyme disease, diphtheria, and leprosy, (9) inflammatory disorders, (10) peripheral nerve disorders (e.g., neuromas), (11) hereditary or emerging hereditary disorders (e.g., Charcot-Marie-Tooth disorder), (12) mononeuropathy, (13) polyneuropathy, or a combination thereof. In some embodiments, the neuropathic pain is caused by or associated with diabetes (type I or type II). In some embodiments, the neuropathic pain is diabetic peripheral neuropathy.

[0144] In some embodiments, neuropathic pain is caused by or is associated with exposure to infectious agents (including, for example, tick-borne infections, varicella zoster, Epstein-Barr virus, West Nile virus, cytomegalovirus, herpes simplex virus, AIDS), or exposure to toxic agents (e.g., drugs, alcohol, heavy metals (e.g., lead, arsenic, mercury), or exposure to industrial agents (e.g., solvents, fumes from glues) and nitrous oxide).

[0145] In some embodiments, neuropathic pain is caused or associated with physical injury, infection, diabetes, cancer therapy, alcoholism, amputation, multiple sclerosis, herpes zoster, spinal surgery, sciatica (pain along the sciatic nerve), low back pain, neuralgia such as trigeminal neuralgia (e.g., pain in the trigeminal region of the face or mouth), neuropathic pain such as painful polyneuropathy (e.g., foot pain, which can extend to include the calf, thigh and hand), or a combination thereof. In some embodiments, neuropathic pain is trigeminal neuralgia. In some embodiments, neuropathic pain is associated with muscle weakness in the back, legs, buttocks or face. In some embodiments, neuropathic pain is caused by compression of nerves (e.g., nerves of the legs, feet, buttocks or nerves of facial muscles). In some embodiments, neuropathic pain includes sciatic nerve injury. In some embodiments, neuropathic pain is sciatica.

[0146] In some embodiments, the method of the present disclosure can reverse, alleviate, improve, inhibit, slow down or prevent one or more symptoms associated with neuropathic pain. Therefore, in one aspect, the present disclosure provides a method for improving the hyperalgesia of a subject in need thereof, including administering an antagonist for FAM19A5 to the subject. As used herein, the term "hyperalgesia" refers to an increased or aggravated reaction to painful stimulation (e.g., acupuncture or hot plate). In some embodiments, hyperalgesia is directed to mechanical stimulation, such as acupuncture (mechanical hyperalgesia). In other embodiments, hyperalgesia is directed to thermal stimulation, such as hot plate (thermal hyperalgesia). In some embodiments, a subject in need thereof suffers from chronic contractile injury (e.g., sciatica). In some embodiments, a subject in need thereof suffers from diabetic peripheral neuropathy.

[0147] In some embodiments, compared with a reference control (e.g., a neuropathic pain subject who has not received a FAM19A5 antagonist), applying a FAM19A5 antagonist to a subject in need allows the subject to have a higher threshold value for mechanical stimulation. As used herein, the term "threshold value for mechanical stimulation" refers to the amount of pressure (from mechanical stimulation) before the subject reacts to the stimulus (e.g., by pulling away). Therefore, compared with a subject with a lower threshold value, a subject with a higher threshold value can withstand or resist a larger amount of mechanical stimulation. In some embodiments, compared with a reference control (e.g., a subject's threshold value before applying a FAM19A5 antagonist), methods disclosed herein can increase the subject's threshold value for mechanical stimulation by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 150% or at least 200%.

[0148] In some embodiments, administration of a FAM19A5 antagonist to a subject in need thereof increases the subject's latency to a thermal stimulus (e.g., a hot plate) (i.e., the time interval between stimulation and response) compared to a reference control (e.g., a neuropathic pain subject who has not received a FAM19A5 antagonist). In some embodiments, the methods disclosed herein can increase the subject's latency to a thermal stimulus by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 150%, or at least 200% compared to a reference control (e.g., the subject's threshold before administration of the FAM19A5 antagonist).

[0149] In another aspect, the present disclosure provides a method for improving the sensory nerve conduction velocity of a subject in need thereof. As used herein, the term "sensory nerve conduction velocity" (SNCV) refers to the rate at which electrical signals travel through peripheral nerves. Healthy nerves send electrical signals faster and with greater intensity than damaged nerves. See Chouhan S., J Clin Diagn Res 10(1):CC01-3(2016). Therefore, tests that help measure SNCV (e.g., sensory nerve conduction velocity tests) can be used to identify potential nerve damage and / or dysfunction in subjects. In some embodiments, compared to a reference control (e.g., a threshold value of a subject before administering a FAM19A5 antagonist), the methods disclosed herein can increase the SNCV of a subject with neuropathic pain by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 150%, or at least 200%.

[0150] There are animal models for studying neuropathic pain. Non-limiting examples of such animal models include (1) a spinal nerve ligation (SNL) model, in which one or more spinal nerves leading to the foot are ligated and cut (see Kim SH and Chung JM., Pain 50:355–363 (1992)); (2) a partial sciatic nerve ligation (PSL) model, in which a portion of the sciatic nerve is tightly ligated (see Seltzer et al., Pain 43:205–218 (1990)); (3) a chronic constriction injury (CCI) model, which involves placing four loose chromic gut ligatures on the sciatic nerve, and an immune response to the sutures results in nerve swelling and nerve constriction (see, e.g., Example 6); (4) a sparing nerve injury (SNI) model, in which the common peroneal and tibial nerves are cut, sparing the sural nerve (see Decosterd I and Woolf CJ, Pain 87:149-158 (2000)); and (5) streptozotocin (STZ)-induced diabetic rats, in which injection of STZ causes pancreatic swelling and degeneration of Langerhans islet β cells, thereby causing experimental diabetes in rats. (See, e.g., Example 8; Akbarzadeh A et al., Indian J. Clin. Biochem. 22(2):60-64 (2007)). These models result in hyperalgesia in animals, manifested as enhanced responses to mechanical and / or thermal stimuli.

[0151] Tests for mechanical hyperalgesia in animals include the Von Frey test, in which Von Frey monofilaments with varying bending forces are applied to the plantar surface of the foot. The threshold for paw withdrawal is significantly reduced following nerve injury. Li et al., Pain 85:493–502 (2000); see also Example 6. Thus, in some embodiments, the methods disclosed herein can increase the threshold for paw withdrawal in an animal model of neuropathic pain (e.g., a chronic constriction injury model).

[0152] Testing for thermal (heat) hyperalgesia can include using a radiant heat source (e.g., a hot plate) focused on the plantar surface of the foot and measuring the reaction time of paw withdrawal. After nerve injury, the foot withdraws faster than before injury. Kim SH and Chung JM. Pain 50:355-363 (1992); see also Example 8. In some embodiments, the methods disclosed herein can increase the latency of paw withdrawal in an animal model of neuropathic pain (e.g., a diabetic peripheral neuropathy model).

[0153] In some embodiments, the subject treated with the methods of the present disclosure is a non-human animal, such as a rat or mouse. In some embodiments, the subject treated in the methods is a human.

[0154] In some embodiments, a FAM19A5 antagonist (e.g., an anti-FAM19A5 antibody or antigen-binding portion thereof), bispecific molecule, or immunoconjugate disclosed herein, or a composition thereof, is administered intravenously, orally, parenterally, intrathecally, intracerebroventricularly, intrapulmonaryly, intramuscularly, subcutaneously, intraperitoneally, intravitreally, or intracerebroventricularly.

[0155] In some embodiments, the FAM19A5 antagonist or composition thereof can be administered in combination with one or more additional agents for treating neuropathic pain. For example, non-limiting exemplary agents for treating neuropathic pain include venlafaxine Antiepileptic drugs such as carbamazepine ( FDA-approved for pain relief in trigeminal neuralgia), gabapentin ( Approved to treat postherpetic neuralgia (PHN): pain that lasts one to three months after shingles heals) and pregabalin ( Approved for PHN, painful diabetic neuropathic pain, and fibromyalgia), sodium channel blockers such as lidocaine, adrenergic drugs such as clonidine, phentolamine, phenoxybenzamine, reserpine, dexmedetomidine, and antidepressants such as amitriptyline, imipramine, and duloxetine.

[0156] Dosages and administration of one or more additional therapeutic agents are known in the art, for example, as indicated by the product labeling of the corresponding drug.

[0157] III. FAM19A5 antagonists

[0158] One or more FAM19A5 antagonists can be used to treat neuropathic pain. In some embodiments, the FAM19A5 antagonist is an antisense oligonucleotide, siRNA, shRNA, miRNA, dsRNA, aptamer, PNA (peptide nucleic acid) or a vector comprising them that specifically targets FAM19A5. In other embodiments, the FAM19A5 antagonist is an antibody or an antigen-binding fragment thereof that specifically binds to the FAM19A5 protein, a polynucleotide encoding an anti-FAM19A5 antibody or an antigen-binding fragment thereof, or a vector comprising its polynucleotide.

[0159] Antibodies available in the methods disclosed herein 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 soluble human FAM 19A5 with a KD of 10 nM or less; (b) bind to membrane-bound human FAM19A5 with a KD of 10 nM or less; or both (a) and (b).

[0160] In some embodiments, the anti-FAM19A5 antibody or its antigen-binding portion binds to the antigen with high affinity (e.g., 10-7 M or less, 10-8 M or less, 10-9 M (1 nM) or less, 10-10 M (0.1 nM) or less, 10-11 M or less, or 10-12 M or less, such as 10-12 M to 10-7 M, 10-11 M to 10-7 M, 10-10 M To 10-7M, or 10-9M to 10-7M, for example, 10-12M, 5×10-12M, 10-11M, 5×10-11M, 10-10M, 5×10-10M, 10-9M, 5×10-9M, 10-8M, 5×10-8M, 10-7M or 5×10-7M KD) specifically binds to soluble human FAM19A5 or membrane-bound human. Standard assays for assessing the binding ability of antibodies to human FAM19A5 of different species are known in the art, including, for example, ELISA, Western blot and RIA. Suitable assays are described in detail in the Examples. The binding kinetics (e.g., binding affinity) of the antibody can also be assessed by standard assays known in the art, such as by ELISA, BIACORE™ analysis or Assays for evaluating the effects of antibodies on the functional properties of FAM19A5 (eg, ligand binding) are described in further detail below and in the Examples.

[0161] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, binds to soluble human FAM19A5 with a KD of 10-7 M or less, 10-8 M (10 nM) or less, 10-9 M (1 nM) or less, 10-10 M or less, 10-12 M to 10-7 M, 10-11 M to 10-7 M, 10-10 M to 10-7 M, 10-9 M to 10-7 M, or 10-8 M to 10-7 M, e.g., as determined by ELISA. In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, binds to soluble FAM19A5 with a KD of 10 nM or less, e.g., between 0.1 and 10 nM, between 0.1 and 5 nM, between 0.1 and 1 nM, between 0.5 and 10 nM, between 0.5 and 5 nM, between 0.5 and 1 nM, between 1 and 10 nM, between 1 and 5 nM, or between 5 and 10 nM. In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, is present at 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, The invention further comprises a KD of about 10 nM, 20 nM, 30 nM, 40 nM, 50 nM, 60 nM, 70 nM, 80 nM, or 90 nM, or about 10 nM, 20 nM, 30 nM, 40 nM, 50 nM, 60 nM, 70 nM, 80 nM, or 90 nM, and specifically binds to soluble human FAM19A5.

[0162] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, binds to membrane-bound human with a KD of 10-7 M or less, 10-8 M (10 nM) or less, 10-9 M (1 nM) or less, 10-10 M or less, 10-12 M to 10-7 M, 10-11 M to 10-7 M, 10-10 M to 10-7 M, 10-9 M to 10-7 M, or 10-8 M to 10-7 M, e.g., as determined by ELISA. In certain embodiments, an anti-FAM19A5 antibody, or an antigen-binding portion thereof, specifically binds to membrane-bound human FAM19A5 with a KD of 10 nM or less, e.g., between 0.1 and 10 nM, between 0.1 and 5 nM, between 0.1 and 1 nM, between 0.5 and 10 nM, between 0.5 and 5 nM, between 0.5 and 1 nM, between 1 and 10 nM, between 1 and 5 nM, or between 5 and 10 nM as determined by ELISA. In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, is present at 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, 10 nM, 20 nM, 30 nM, 40 nM, 50 nM, 60 nM, 70 nM, 80 nM, or 90 nM, or a KD of about 10 nM, 20 nM, 30 nM, 40 nM, 50 nM, 60 nM, 70 nM, 80 nM, or 90 nM.

[0163] In some embodiments, an anti-FAM19A5 antibody, or antigen binding portion thereof, suitable for the methods disclosed herein cross-competes for binding to (or inhibits binding to) a human FAM19A5 epitope with an anti-FAM19A5 antibody comprising a CDR or variable region disclosed herein.

[0164] In some embodiments, the anti-FAM19A5 antibody, or an 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, respectively; (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, and 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, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: NO:28 amino acid sequence; (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, and 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: 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, 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, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:151 NO: 151, the light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 152, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 153, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 154. In some embodiments, the heavy chain CDR1, CDR2 and CDR3 of the reference antibody comprise the CDR1, CDR2 and CDR3 sequences as set forth in Table 2, respectively, and the light chain CDR1, CDR2 and CDR3 of the reference antibody comprise the CDR1, CDR2 and CDR3 sequences as set forth in Table 3, respectively. ;

[0165] In some embodiments, 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. (k) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 161 and 172, respectively; (l) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 162 and 173, respectively; (m) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 163 and 174, respectively; (n) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 164 and 175, respectively; or (o) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 165 and 176, respectively. In some embodiments, the reference antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises an amino acid sequence as set forth in Table 4, and the VL comprises an amino acid sequence as set forth in Table 5.

[0166] In some embodiments, the anti-FAM19A5 antibody or its antigen binding portion inhibits the binding of such a reference antibody to human FAM19A5 by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%. The competing antibodies bind to the same epitope, overlapping epitopes or adjacent epitopes (e.g., as 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.

[0167] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, binds to the same FAM19A5 epitope as a reference antibody disclosed herein, the reference antibody 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, and 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, and 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, and 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: The heavy chain CDR2 comprises the amino acid sequence of SEQ ID 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, 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, the heavy chain CDR3 comprises the amino acid sequence of SEQ ID NO:151 The light chain CDR1 comprises the amino acid sequence of SEQ ID NO: 152, the light chain CDR2 comprises the amino acid sequence of SEQ ID NO: 153, and the light chain CDR3 comprises the amino acid sequence of SEQ ID NO: 154. ;

[0168] In some embodiments, 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. (k) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 161 and 172, respectively; (l) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 162 and 173, respectively; (m) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 163 and 174, respectively; (n) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 164 and 175, respectively; or (o) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 165 and 176, respectively.

[0169] Techniques for determining whether two antibodies bind to the same epitope include, for example, epitope mapping methods such as X-ray analysis of crystals of the antigen:antibody complex (which provides atomic resolution of the epitope) and hydrogen / deuterium exchange mass spectrometry (HDX-MS), methods of monitoring antibody binding to antigen fragments or mutated variants of the antigen, where loss of binding resulting from modification of amino acid residues in the antigen sequence is generally considered indicative of epitope composition, and computational combinatorial methods for epitope mapping.

[0170] The anti-FAM19A5 antibodies or antigen-binding portions thereof that can be used in the methods disclosed herein can bind to at least one epitope of mature human FAM19A5, as determined by, for example, binding of the antibody to a fragment of human FAM19A5. In some embodiments, the anti-FAM19A5 antibodies or antigen-binding fragments thereof bind to a fragment located within the amino acid sequence of TLDRDSSQPRRTIARQTARC (amino acid residues 42 to 61 of SEQ ID NO: 6 or SEQ ID NO: 2), for example, an epitope having at least 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: 6. In some embodiments, an anti-FAM19A5 antibody or antigen-binding portion thereof binds to SEQ ID NO: 6 at one or more amino acids corresponding to amino acid residues 46 to 51 (ie, DSSQPR), such as amino acid residues 46, 50 and 52 (ie, D---PR), such as amino acid residues 46, 47, 48 and 50 (ie, DSS-P) of SEQ ID NO: 2. In some embodiments, an anti-FAM19A5 antibody or antigen-binding fragment thereof binds to a fragment located within the amino acid sequence of CDMLPCLEGEGCDLLINRSG (amino acids 90 to 109 of SEQ ID NO: 9 or SEQ ID NO: 2), such as an epitope having at least 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: 9. In certain embodiments, an anti-FAM19A5 antibody, or an antigen-binding portion thereof, binds to SEQ ID NO: 9 at one or more amino acid residues 99 to 107 of SEQ ID NO: 4 (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).

[0171] In some embodiments, 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 some embodiments, 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.

[0172] In some embodiments, the anti-FAM19A5 antibody, or an antigen binding portion thereof, binds only to a human FAM19A5 epitope that is 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 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.

[0173] In some embodiments, 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 configuration (i.e., undenatured). In some embodiments, the anti-FAM19A5 antibodies or antigen binding fragments thereof of the present disclosure bind to SEQ ID NO: 9 or a fragment thereof in its native configuration (i.e., undenatured). In some embodiments, the anti-FAM19A5 antibodies or antigen binding portions thereof of the present disclosure bind to glycosylated and unglycosylated human FAM19A5.

[0174] In some embodiments, the anti-FAM19A5 antibody or its antigen binding portion further binds to one or more additional FAM19A5 epitopes. Thus, certain anti-FAM19A5 antibodies or their antigen binding portions bind to (i) an epitope of SEQ ID NO:6 and an additional epitope, or (ii) an epitope of SEQ ID NO:9 and an additional epitope. Other anti-FAM19A5 antibodies or their antigen binding fragments may bind to an epitope of SEQ ID NO:5, SEQ ID NO:9 and an additional epitope. In some embodiments, an anti-FAM19A5 antibody or its antigen binding fragment binds to an epitope of SEQ ID NO:6, an epitope of SEQ ID NO:10 and an additional epitope.

[0175] In some embodiments, 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 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 include 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 and SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10. In some embodiments, one or more additional FAM19A5 epitopes are selected from SEQ ID NO: 5, 6, 7, 8, 9 or 10, or fragments located within the amino acid sequence of SEQ ID NO: 5, 6, 7, 8, 9 or 10, such as 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 SEQ ID NO: 5, 6, 7, 8, 9 or 10, or any combination thereof. In some embodiments, the anti-FAM19A5 antibodies or antigen binding portions thereof of the present disclosure bind to any of the one or more additional epitopes in their native configuration (i.e., undenatured). In some embodiments, the anti-FAM19A5 antibodies or antigen binding portions thereof bind to one or more additional glycosylated and non-glycosylated FAM19A5 epitopes.

[0176] In some embodiments, 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), and wherein EP8 comprises, consists essentially of, or consists of the amino acids TCTQPGGR (SEQ ID NO: 72). In some embodiments, 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 some embodiments, an anti-FAM19A5 antibody, or an antigen binding portion thereof, binds only to EP2. In some embodiments, an anti-FAM19A5 antibody, or an antigen binding portion thereof, binds to EP4 and EP8.

[0177] In some embodiments, 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), and wherein EP8 comprises the amino acids TCTQPGGR (SEQ ID NO: 72). In some embodiments, 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 some embodiments, an anti-FAM19A5 antibody, or an antigen binding portion thereof, binds only to EP6, EP7, or EP8. In some embodiments, an anti-FAM19A5 antibody, or an antigen binding portion thereof, binds to EP6, EP7, and EP8. In some embodiments, an anti-FAM19A5 antibody, or an antigen binding portion thereof, binds to EP7 and EP8. In some embodiments, the anti-FAM19A5 antibody, or antigen binding portion thereof, binds to EP7.

[0178] In some embodiments, the anti-FAM19A5 antibody, or antigen binding portion thereof, binds to 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.

[0179] In some embodiments, provided herein is an antibody or an antigen binding portion thereof, which binds to FAM19A5 (e.g., human FAM19A5) with 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or higher affinity compared to another protein in the FAM19A family, the affinity is such as measured by, for example, immunoassay (e.g., ELISA), surface plasmon resonance or kinetic exclusion assay. In a specific embodiment, provided herein is an antibody or an antigen binding portion thereof that is bound to FAM19A5 (e.g., human FAM19A5) that has no cross-reactivity with another protein in the FAM19A family, such as measured by, for example, immunoassay.

[0180] In some embodiments, 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, such as having more, less or a different type of post-translational modification.

[0181] IV. Exemplary Anti-FAM19A5 Antibodies

[0182] Specific antibodies that can be used in the methods disclosed herein are antibodies (e.g., monoclonal antibodies) having the CDR and / or variable region sequences of antibodies 1-65 isolated in Example 1, 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. The amino acid sequences of the VH and VL CDRs of different anti-FAM19A5 antibodies are provided in Tables 2 and 3, respectively. The CDRs of the following antibodies were identified 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 using the IMGT numbering system (see above): 1-7A-IT, low-PI, 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. The VH and VL amino acid sequences of different anti-FAM19A5 antibodies of the present disclosure are provided in Tables 4 and 5, respectively.

[0183] Table 2 Variable heavy chain CDR amino acid sequences

[0184]

[0185]

[0186]

[0187] Table 3 Variable light chain CDR amino acid sequences

[0188]

[0189]

[0190]

[0191] Table 4: Variable heavy chain amino acid sequences

[0192]

[0193]

[0194]

[0195] Table 5: Variable light chain amino acid sequences

[0196]

[0197]

[0198] In some embodiments, the anti-FAM19A5 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NOs: 35-38, 155-165, 203, 223-226, 254, 257, 271 or 275. In other embodiments, the isolated anti-FAM19A5 antibody or antigen-binding portion thereof comprises a CDR of the heavy chain variable region selected from the group consisting of: SEQ ID NOs: 35-38, 155-165, 203, 223-226, 254, 257, 271 or 275.

[0199] In some embodiments, the isolated anti-FAM19A5 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NOs: 39-42, 166-176, 204, 227-234 or 272. In other embodiments, the isolated anti-FAM19A5 antibody or antigen-binding portion thereof comprises a CDR of the light chain variable region selected from the group consisting of: SEQ ID NOs: 39-42, 166-176, 227-234 or 272.

[0200] In some embodiments, the isolated anti-FAM19A5 antibody, or antigen binding portion thereof, comprises CDRs of a heavy chain variable region selected from the group consisting of: SEQ ID NOs: 35-38, 155-165, 203, 223-226, 254, 257, 271, or 275 and CDRs of a light chain variable region selected from the group consisting of: SEQ ID NOs: 39-42, 166-176, 227-234, or 272.

[0201] In some embodiments, the isolated anti-FAM19A5 antibody or antigen-binding fragment thereof comprises 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: 37 and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 41; (ii) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 36 and 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: 35 and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 39; (iv) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 38 and 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 and 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: 156 and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: NO:167; (vii) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:157, and 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, and 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, and 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, and 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, and 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:162, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: NO:173; (xiii) wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:163, and 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, and 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, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:176.In some embodiments, the isolated anti-FAM19A5 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein VH comprises a VH sequence as set forth in Table 4, and VL comprises a VL sequence as set forth in Table 5.

[0202] In some embodiments, the isolated anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises 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 shown in SEQ ID NOs: 35-38, 155-165, 203, 223-226, 254, 257, 271 or 275.

[0203] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises 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 shown in SEQ ID NOs: 39-42, 166-176, 227-234 or 272.

[0204] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises 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 NOs:35-38, 155-165, 203, 223-226, 254, 257, 271 or 275, and 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 NOs:39-42, 166-176, 227-234 or 272.

[0205] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises:

[0206] (a) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 35 and 39, respectively;

[0207] (b) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 36 and 40, respectively;

[0208] (c) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 37 and 41, respectively;

[0209] (d) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 38 and 42, respectively;

[0210] (e) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 155 and 166, respectively;

[0211] (f) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 156 and 167, respectively;

[0212] (g) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 157 and 168, respectively;

[0213] (h) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 158 and 169, respectively;

[0214] (i) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 159 and 170, respectively;

[0215] (j) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 160 and 171, respectively;

[0216] (k) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 161 and 172, respectively;

[0217] (l) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 162 and 173, respectively;

[0218] (m) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 163 and 174, respectively;

[0219] (n) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 164 and 175, respectively; or

[0220] (o) heavy chain and light chain variable region sequences comprising SEQ ID NOs: 165 and 176, respectively.

[0221] In certain embodiments, an anti-FAM19A5 antibody or antigen-binding portion thereof comprises (i) a heavy chain CDR1, CDR2 and CDR3 of 1-65 or a combination thereof, and / or a light chain CDR1, CDR2 and CDR3 of 1-65 or a combination thereof; (ii) a heavy chain CDR1, CDR2 and CDR3 of 3-2 or a combination thereof, and / or a light chain CDR1, CDR2 and CDR3 of 3-2 or any combination thereof; (iii) a heavy chain CDR1, CDR2 and CDR3 of 2-13 or a combination thereof, and / or a light chain CDR1, CDR2 and CDR3 of 2-13 or any combination thereof; (iv) a heavy chain CDR1, CDR2 and CDR3 of 1-28 or a combination thereof. R3 or a combination thereof, and / or the light chain CDR1, CDR2 and CDR3 of 1-28 or any combination thereof; (v) the heavy chain CDR1, CDR2 and CDR3 of P2-C12 or a combination thereof, and / or the light chain CDR1, CDR2 and CDR3 of P2-C12 or any combination thereof; (vi) the heavy chain CDR1, CDR2 and CDR3 of 13B4 or a combination thereof, and / or the light chain CDR1, CDR2 and CDR3 of 13B4 or any combination thereof; (vii) the heavy chain CDR1, CDR2 and CDR3 of 13F7 or a combination thereof, and / or the light chain CDR1, CDR2 and CDR3 of 13F7 or any combination thereof; viii) the heavy chain CDR1, CDR2 and CDR3 or any combination thereof of 15A9, 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's light chain CDR1, CDR2 and CDR3 or any combination thereof; (xii) P2-A01's heavy chain CDR1, CDR2 and CDR3 or any combination thereof, and / or P2-A01's light chain CDR1, CDR2 and CDR3 or any combination thereof; (xiii) P2-A03's heavy chain CDR1, CDR2 and CDR3 or any combination thereof, and / or P2-A03's light chain CDR1, CDR2 and CDR3 or any combination thereof; (xiv) P2-F07's heavy chain CDR1, CDR2 and CDR3 or any combination thereof, and / or P2-F07's light chain CDR1, CDR2 and CDR3 or any combination thereof;or (xv) the heavy chain CDR1, CDR2 and CDR3 of P2-F11 or a combination thereof, and / or the light chain CDR1, CDR2 and CDR3 of F2-F11 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 VL CDR1, CDR2 and CDR3 of different anti-FAM19A5 antibodies disclosed herein are provided in Table 3. ;

[0222] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises:

[0223] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 17; and / or

[0224] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 18; and / or

[0225] (c) VH CDR3 comprising the amino acid sequence of SEQ ID NO:19.

[0226] In some embodiments, an anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises one, two, or all three of the above-described VH CDRs.

[0227] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises:

[0228] (a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 29; and / or

[0229] (b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 30; and / or

[0230] (c) VL CDR3 comprising the amino acid sequence of SEQ ID NO:31.

[0231] In some embodiments, an anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises one, two, or all three of the above-described VL CDRs.

[0232] In some embodiments, the anti-FAM19A5 antibody, or antigen-binding portion thereof, comprises:

[0233] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 17;

[0234] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 18;

[0235] (c) a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 19;

[0236] (d) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 29;

[0237] (e) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 30; and / or

[0238] (f) VL CDR3 comprising the amino acid sequence of SEQ ID NO:31.

[0239] In some embodiments, the anti-FAM19A5 antibody or antigen-binding fragment thereof comprises:

[0240] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 14;

[0241] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 15; and / or

[0242] (c) VH CDR3 comprising the amino acid sequence of SEQ ID NO:16.

[0243] In some embodiments, an anti-FAM19A5 antibody or antigen-binding fragment thereof comprises one, two, or all three of the above-mentioned VH CDRs.

[0244] In some embodiments, the anti-FAM19A5 antibody or antigen-binding fragment thereof comprises:

[0245] (a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 26;

[0246] (b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 27; and / or

[0247] (c) VL CDR3 comprising the amino acid sequence of SEQ ID NO:28.

[0248] In some embodiments, an anti-FAM19A5 antibody or antigen-binding fragment thereof comprises one, two, or all three of the above-described VL CDRs.

[0249] In some embodiments, the anti-FAM19A5 antibody or antigen-binding fragment thereof comprises:

[0250] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 14;

[0251] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 15;

[0252] (c) a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 16;

[0253] (d) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 26;

[0254] (e) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 27; and / or

[0255] (f) VL CDR3 comprising the amino acid sequence of SEQ ID NO:28.

[0256] In some embodiments, an anti-FAM19A5 antibody or antigen-binding fragment thereof that specifically binds to human FAM19A5 comprises:

[0257] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 11;

[0258] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 12; and / or

[0259] (c) VH CDR3 comprising the amino acid sequence of SEQ ID NO:13.

[0260] In some embodiments, the antibody or antigen-binding fragment thereof comprises one, two, or all three of the above-mentioned VH CDRs.

[0261] In some embodiments, an anti-FAM19A5 antibody or antigen-binding fragment thereof that specifically binds to human FAM19A5 comprises:

[0262] (a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 23;

[0263] (b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 24; and / or

[0264] (c) VL CDR3 comprising the amino acid sequence of SEQ ID NO:25.

[0265] In some embodiments, the antibody or antigen-binding fragment thereof comprises one, two, or all three of the above-described VL CDRs.

[0266] In some embodiments, an anti-FAM19A5 antibody or antigen-binding fragment thereof that specifically binds to human FAM19A5 comprises:

[0267] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 11;

[0268] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 12;

[0269] (c) a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 13;

[0270] (d) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 23;

[0271] (e) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 24; and / or

[0272] (f) VL CDR3 comprising the amino acid sequence of SEQ ID NO:25.

[0273] In some embodiments, the anti-FAM19A5 antibody or antigen-binding fragment thereof comprises:

[0274] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 20;

[0275] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 21; and / or

[0276] (c) VH CDR3 comprising the amino acid sequence of SEQ ID NO:22.

[0277] In some embodiments, an anti-FAM19A5 antibody or antigen-binding fragment thereof comprises one, two, or all three of the above-mentioned VH CDRs.

[0278] In some embodiments, the anti-FAM19A5 antibody or antigen-binding fragment thereof comprises:

[0279] (a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 32;

[0280] (b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 33; and / or

[0281] (c) VL CDR3 comprising the amino acid sequence of SEQ ID NO:34.

[0282] In some embodiments, an anti-FAM19A5 antibody or antigen-binding fragment thereof comprises one, two, or all three of the above-described VL CDRs.

[0283] In some embodiments, the anti-FAM19A5 antibody or antigen-binding fragment thereof comprises:

[0284] (a) a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 20;

[0285] (b) a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 21;

[0286] (c) a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 22;

[0287] (d) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 32;

[0288] (e) a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 33; and / or

[0289] (f) VL CDR3 comprising the amino acid sequence of SEQ ID NO:34.

[0290] In some embodiments, an anti-FAM19A5 antibody, or antigen-binding portion thereof, disclosed herein comprises one, two, three, four, five, or six of the above-mentioned CDRs.

[0291] The VH domains described herein or one or more of their CDRs can be connected to a constant domain for forming 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 connected to a constant domain for forming 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.

[0292] Therefore, in a specific embodiment, an antibody comprising an antibody light chain and a heavy chain (e.g., a separated light chain and a heavy chain) that can be used for the methods disclosed herein is provided. With regard to the light chain, in a specific embodiment, the light chain of the antibody described herein is a κ light chain. In another specific embodiment, the light chain of the antibody described herein is a λ light chain. In another specific embodiment, the light chain of the antibody described herein is a human κ light chain or a human λ light chain. In a specific embodiment, the antibody specifically bound to the FAM19A5 polypeptide (e.g., human FAM19A5) described in the methods disclosed herein comprises a light chain, the light chain comprises any VL or VL CDR amino acid sequence described herein, and wherein the constant region of the light chain comprises the amino acid sequence of the human κ light chain constant region. In a specific embodiment, the antibody specifically bound to the FAM19A5 polypeptide (e.g., human FAM19A5) described in the methods disclosed herein comprises a light chain, the light chain comprises a VL or VL CDR amino acid sequence described herein, and wherein the constant region of the light chain comprises the amino acid sequence of the human λ light chain constant region. Non-limiting examples of human constant region sequences have been described in the art, see, e.g., U.S. Pat. No. 5,693,780 and Kabat EA et al., (1991) supra.

[0293] With regard to the heavy chain, in some embodiments, the heavy chain of the antibody described herein may be an alpha (α), delta (δ), epsilon (ε), gamma (γ) or mu (μ) heavy chain. In another specific embodiment, the heavy chain of the antibody described may comprise a human alpha (α), delta (δ), epsilon (ε), gamma (γ) or mu (μ) heavy chain. In some embodiments, the antibody specifically binding to FAM19A5 (e.g., human FAM19A5) described in the methods disclosed herein comprises a heavy chain comprising a VH or VH CDR amino acid sequence described herein, and wherein the constant region of the heavy chain comprises the amino acid sequence of a human gamma (γ) heavy chain constant region. In other embodiments, the antibody described herein that specifically binds to FAM19A5 (e.g., human FAM19A5) comprises a heavy chain comprising a VH or VH CDR amino acid sequence disclosed herein, and wherein the constant region of the heavy chain comprises the amino acid of a human heavy chain described herein or known in the art. Non-limiting examples of human constant region sequences have been described in the art, see, eg, US Pat. No. 5,693,780 and Kabat EA et al., (1991) supra.

[0294] In some embodiments, the antibodies described herein that specifically bind to FAM19A5 (e.g., human FAM19A5) that can be used in the methods disclosed herein include a VL domain and a VH domain, the domains including VH or VH CDRs and VL and VL CDRs as described herein, and wherein the constant region includes an IgG, IgE, IgM, IgD, IgA or IgY immunoglobulin molecule or an amino acid sequence of a constant region of a human IgG, IgE, IgM, IgD, IgA or IgY immunoglobulin molecule. In another specific embodiment, the antibodies described herein that specifically bind to FAM19A5 (e.g., human FAM19A5) include a VL domain and a VH domain, which include any amino acid sequence described herein, and wherein the constant region includes an amino acid sequence of a constant region of an IgG, IgE, IgM, IgD, IgA or IgY immunoglobulin molecule, any subclass of an immunoglobulin molecule (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2). In some embodiments, the constant region comprises the amino acid sequence of the constant region of naturally occurring human IgG, including subclasses (e.g., IgG1, IgG2, IgG3, or IgG4) and allotypes (e.g., G1m, G2m, G3m, and nG4m) and variants thereof. See, e.g., Vidarsson G et al. Front Immunol. 5:520 (published online on October 20, 2014) and Jefferis R and Lefranc MP, mAbs 1:4, 1-7 (2009). In some embodiments, the constant region comprises the amino acid sequence of the constant region of human IgG1, IgG2, IgG3, or IgG4, or variants thereof.

[0295] In certain embodiments, the disclosed anti-FAM19A5 antibodies or antigen-binding portions thereof that can be used in the methods disclosed herein 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 largely overlapping binding site for C1q (complement) and IgG-Fc receptors (FcγR) on effector cells of the innate immune system. Moreover, IgG2 and IgG4 antibodies have lower levels of Fc effector functions than 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 an antibody fragment lacking an Fc region (e.g., such as Fab, F(ab')2, single-chain Fv (scFv), or sdAb composed of a monomeric VH or VL domain); (2) producing an aglycosylated antibody, which can be produced by, for example, deleting or changing the residue to which the sugar is attached, enzymatically removing the sugar, producing the antibody in cells cultured in the presence of a glycosylation inhibitor, or by expressing the antibody in cells that are unable to glycosylate the protein (e.g., bacterial host cells, see, e.g., U.S. Patent Publication No. 20120100140); (3) employing 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, see, e.g., U.S. Patent Publication Nos. 20120100140 and Lau et al.); C et al. J. Immunol. 191:4769-4777 (2013)); and (4) generating an Fc region with a mutation resulting in reduced or no Fc function. See, e.g., U.S. Patent Publication No. 20120100140 and the U.S. and PCT applications cited therein and An et al., mAbs 1:6,572-579 (2009).

[0296] Thus, in some embodiments, the anti-FAM19A5 antibodies or antigen-binding portions thereof useful in the methods disclosed herein 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.

[0297] In some embodiments, the anti-FAM19A5 antibody or antigen-binding portion thereof that can be used in the methods disclosed herein comprises an Fc region with reduced Fc effector function or without Fc effector function. In some embodiments, the constant region comprises the amino acid sequence of the Fc region of human IgG2 or IgG4, and in some embodiments, the anti-FAM19A5 antibody is an IgG2 / IgG4 isotype. In some embodiments, the anti-FAM19A5 antibody comprises: a chimeric Fc region comprising a CH2 domain of an IgG antibody from an IgG4 isotype and a CH3 domain of an IgG antibody from an IgG1 isotype; or a chimeric Fc region comprising a hinge region from IgG2 and a CH2 region from IgG4; or an Fc region with a mutation that results in reduced Fc function or without Fc function. Fc regions with reduced Fc effector function or without Fc effector function include those known in the art. See, e.g., Lau C. et al., J. Immunol. 191: 4769-4777 (2013); An et al., mAbs 1: 6,572-579 (2009); and U.S. Patent Publication No. 20120100140 and the U.S. patents and publications and PCT publications cited therein. One of ordinary skill in the art can also easily prepare an Fc region with reduced or no Fc effector function.

[0298] V. Nucleic Acid Molecules

[0299] Another aspect described herein relates to one or more nucleic acid molecules, which encode any of the antibodies or antigen binding portions thereof that can be used in the methods disclosed herein. Nucleic acids can exist in intact cells, cell lysates, or partially purified or substantially pure forms. When purified from other cell components or other contaminants (e.g., other cell nucleic acids (e.g., other chromosomal DNA, e.g., chromosomal DNA connected to isolated DNA in nature) or proteins by standard techniques (including alkaline / SDS treatment, CsCl banding, column chromatography, restriction enzymes, agarose gel electrophoresis, and other techniques known in the art), nucleic acids are "separated" or "become substantially pure". See, F.Ausubel et al., ed. (1987) Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York. Nucleic acids described herein can be, for example, DNA or RNA, and may or may not contain intron sequences. In certain embodiments, nucleic acids are cDNA molecules.

[0300] Nucleic acids 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 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.

[0301] Some nucleic acid molecules described herein are those encoding the VH and VL sequences of anti-FAM19A5 antibodies disclosed herein. The exemplary DNA sequences encoding the VH sequences of such antibodies are shown in SEQ ID NOs: 43-46, 177, 205, 235-243, 255, 258, 273 and 276 (see Table 6). The exemplary DNA sequences encoding the VL sequences of such antibodies are shown in SEQ ID NOs: 47-50, 178, 206, 244-252, 256, 259, 274 and 277 (see Table 7).

[0302] Table 6: Variable heavy chain polynucleotide sequences

[0303]

[0304]

[0305]

[0306]

[0307]

[0308] Table 7: Variable light chain polynucleotide sequences

[0309]

[0310]

[0311]

[0312]

[0313] The method for preparing an anti-FAM19A5 antibody or an antigen-binding fragment thereof as disclosed herein may include expressing the relevant heavy and light chains of the antibody in a cell line comprising nucleotide sequences encoding the heavy and light chains with a signal peptide, for example, 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.

[0314] Once the DNA fragments encoding the VH and VL segments are obtained, these DNA fragments can be further manipulated by standard recombinant DNA techniques, for example to convert the variable region genes 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. The term "operably linked" as used in this context means that the two DNA fragments are linked together so that the amino acid sequences encoded by the two DNA fragments remain in frame.

[0315] By operably linking the DNA encoding VH to another DNA molecule encoding the heavy chain constant region (hinge, CH1, CH2 and / or CH3), the separated DNA encoding the VH region can be converted into a full-length heavy chain gene. The sequence of the human heavy chain constant region gene is known in the art (see, e.g., Kabat, EA et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242), and the DNA fragments comprising these regions can be obtained by standard PCR amplification. The heavy chain constant region can be IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM or IgD constant region, such as IgG2 and / or IgG4 constant region. For Fab fragment heavy chain genes, the DNA encoding VH can be operably linked to another DNA molecule encoding only the heavy chain CH1 constant region.

[0316] By operably connecting the DNA encoding VL to another DNA molecule encoding the constant region CL of the light chain, the DNA encoding the separation of the VL region can be converted into a full-length light chain gene (and a Fab light chain gene). The sequence of the human light chain constant region gene is known in the art (see, e.g., Kabat, EA et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USDepartment of Health and Human Services, NIH Publication No.91-3242), and the DNA fragments comprising these regions can be obtained by standard PCR amplification. The constant region of the light chain can be a kappa or lambda constant region.

[0317] To generate an scFv gene, the DNA fragments encoding VH and VL are operably linked to another fragment encoding a flexible linker (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 in which the VL and VH regions are connected by a flexible linker (see, e.g., 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).

[0318] In some embodiments, the present 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 embodiments, the vector can be used for gene therapy.

[0319] Suitable vectors for use in the present disclosure include expression vectors, viral vectors, and plasmid vectors. In some embodiments, the vector is a viral vector.

[0320] As used herein, an expression vector refers to any nucleic acid construct that contains the necessary elements for transcription and translation of an inserted coding sequence, or in the case of an RNA viral vector, contains the necessary elements for replication and translation when introduced into a suitable host cell. Expression vectors can include plasmids, phagemids, viruses and derivatives thereof.

[0321] VI. Antibody Production

[0322] The anti-FAM19A5 antibodies or antigen-binding fragments thereof disclosed herein can be produced by any method known in the art for synthesizing antibodies, such as by chemical synthesis or by recombinant expression technology. Unless otherwise indicated, the methods described herein employ conventional techniques in the relevant fields of molecular biology, microbiology, genetic analysis, recombinant DNA, organic chemistry, biochemistry, PCR, oligonucleotide synthesis and modification, nucleic acid hybridization, and art technology. These techniques are described in, for example, 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, Second Edition, Cold Spring Harbor Laboratory Press; Sambrook J et al., (2001) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; 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 Approach, IRL Press; 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.

[0323] In some embodiments, antibodies useful in the methods disclosed herein are antibodies (e.g., recombinant antibodies) prepared, expressed, produced, or isolated by any means involving, for example, synthesis, genetic engineering of a DNA sequence. In certain embodiments, such antibodies comprise sequences (e.g., DNA sequences or amino acid sequences) that are not naturally present in the antibody germline repertoire in an animal or mammal (e.g., human).

[0324] VII. Pharmaceutical Compositions

[0325] Provided herein are compositions comprising antibodies or antigen-binding portions thereof useful in the methods described herein, the compositions having the desired purity in a physiologically acceptable carrier, excipient or stabilizer (Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, PA). Acceptable carriers, excipients or stabilizers are nontoxic to recipients at the dosages and concentrations employed, 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 and 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).

[0326] In a specific embodiment, the pharmaceutical composition comprises an antibody or antigen binding portion thereof, a bispecific molecule or an immunoconjugate as described herein, and optionally one or more additional prophylactic or therapeutic agents in a pharmaceutically acceptable carrier. In a specific embodiment, the pharmaceutical composition comprises an effective amount of an antibody or antigen binding portion thereof as described herein, and optionally one or more additional prophylactic or therapeutic agents in a pharmaceutically acceptable carrier. In some embodiments, the antibody is the only active ingredient contained in the pharmaceutical composition. The pharmaceutical compositions described herein can be used to enhance, induce or activate FAM19A5 activity and treat conditions such as central nervous system damage, degenerative brain disorders or neuropathic pain.

[0327] The pharmaceutically acceptable carrier used in parenteral preparations includes aqueous vehicles, non-aqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending agents and dispersants, emulsifiers, sequestrants or chelating agents and other pharmaceutically acceptable substances. The example of aqueous vehicles includes 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, cottonseed oil, corn oil, sesame oil and peanut oil of plant origin. Antimicrobial agents of antibacterial or antifungal concentrations can be added to parenteral preparations packaged in multi-dose containers, and the multi-dose containers include phenol or cresol, mercury, benzyl alcohol, chlorobutanol, methylparaben and propylparaben, thimerosal, benzalkonium chloride and benzethonium chloride. Isotonic agents include sodium chloride and dextrose. Buffers include phosphate and citrate. 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). Metal ion sequestrants or chelators 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.

[0328] Pharmaceutical compositions can be formulated for any route of administration to a subject. Specific examples of routes of administration include intranasal, oral, parenteral, intrathecal, intracerebroventricular, pulmonary, subcutaneous, intraperitoneal, intravitreal or intraventricular. Parenteral administration characterized by subcutaneous, intramuscular or intravenous injection is also contemplated herein. Injectables can be in conventional form (as a liquid solution or suspension), in a solid form suitable for dissolving or suspending in a liquid before injection, or prepared as an emulsion. Injectables, solutions and emulsions also contain one or more excipients. Suitable excipients are, for example, water, saline, dextrose, glycerol or ethanol. In addition, if desired, the pharmaceutical composition to be administered can also contain a small amount of non-toxic auxiliary substances, such as wetting agents or emulsifiers, pH buffers, stabilizers, solubility enhancers and other such agents, such as, for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate and cyclodextrin.

[0329] Preparations for parenteral administration of antibodies include sterile solutions ready for injection, sterile dry soluble products (such as lyophilized powders) ready to be mixed with solvents before use, including subcutaneous tablets, sterile suspensions ready for injection, sterile dry insoluble products ready to be mixed with vehicles before use, and sterile emulsions. The solution can be an aqueous solution or a non-aqueous solution.

[0330] 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.

[0331] Topical mixtures containing antibodies are prepared as described for topical and systemic administration. The resulting mixtures may be solutions, suspensions, emulsions, etc., and may be formulated as creams, gels, ointments, emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes, foams, aerosols, rinses, sprays, suppositories, bandages, skin patches, or any other formulation suitable for topical administration.

[0332] The antibodies or antigen-binding portions thereof described herein can be formulated as aerosols for topical administration, such as by inhalation (see, e.g., U.S. Pat. Nos. 4,044,126, 4,414,209, and 4,364,923, which describe aerosols for delivering steroids that can be used to treat inflammatory diseases, particularly asthma). These formulations for administration to the respiratory tract can be in the form of an aerosol or solution for a nebulizer, or as a fine powder for insufflation, alone or in combination with an inert carrier such as lactose. In such cases, in some embodiments, the particles of the formulation will have a diameter of less than 50 microns, in some embodiments less than 10 microns.

[0333] The antibodies or antigen-binding portions thereof described herein can be formulated for local or topical administration in the form of gels, creams and lotions, such as for topical application to the skin and mucous membranes (such as in the eye), and for application to the eye or for intracisternal or intraspinal administration. Topical administration is contemplated for transdermal delivery, and also for application to the eye or mucous membranes, or for inhalation therapy. Nasal solutions of the antibodies can also be administered alone or in combination with other pharmaceutically acceptable excipients.

[0334] Transdermal patches (including iontophoresis and electrophoresis devices) are well known to those skilled in the art and 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.

[0335] In certain embodiments, the pharmaceutical composition comprising an antibody or antigen-binding portion thereof described herein is a lyophilized powder that can be reconstituted for administration in the form of 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 embodiments, the lyophilized powder is sterile. The solvent may contain an excipient that improves the stability of the powder or the reconstituted solution prepared from the powder or other pharmacological components. The excipients that can be used include, but are not limited to, dextrose, sorbitol, fructose, corn syrup, xylitol, glycerol, glucose, sucrose, or other suitable agents. In some embodiments, 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, at a neutral pH. The solution is then sterile filtered and then lyophilized under standard conditions known to those skilled in the art, thereby providing the desired formulation. In some embodiments, the resulting solution will be dispensed 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 at about 4°C to room temperature.

[0336] Reconstitution of this lyophilized powder with water for injection provides a formulation for parenteral administration. For reconstitution, the lyophilized powder is added to sterile water or other suitable carrier. The exact amount depends on the compound selected. Such an amount can be determined empirically.

[0337] Antibodies or antigen-binding portions thereof, bispecific molecules or immunoconjugates described herein and other compositions provided herein can also be formulated into specific tissues, receptors or other regions of the body of a subject to be treated. Many such targeting methods are well known to those skilled in the art. All such targeting methods are contemplated for use in compositions of the present invention 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 embodiment, the antibodies described herein, or antigen-binding portions thereof, are targeted to treat central nervous system damage, degenerative brain disorders, or neuropathic pain.

[0338] Compositions to be used for in vivo administration may be sterile. This is easily achieved by, for example, filtration through sterile filtration membranes.

[0339] VIII. Kit

[0340] Provided herein are kits comprising one or more antibodies or antigen binding portions thereof, bispecific molecules or immunoconjugates thereof that can be used for the methods described herein. In a specific embodiment, provided herein is a pharmaceutical pack or kit comprising one or more containers filled with one or more ingredients of the pharmaceutical compositions described herein (such as one or more antibodies or antigen binding portions thereof provided herein, optional instructions for use). In some embodiments, the kit contains the pharmaceutical compositions described herein and any prophylactic or therapeutic agents, such as those described herein.

[0341] Examples

[0342] The following examples are offered by way of illustration and not limitation.

[0343] Example 1 Expression and purification of human FAM19A5 protein

[0344] Recombinant human FAM19A5 protein is produced and purified as described below, and the purified protein is used for antibody screening assay based on binding affinity analysis. First, the LPS-hT plasmid expressing the FAM19A5 gene is transformed into bacteria, and the inducing protein is overexpressed. Once produced, the FAM19A5 protein is purified using Ni-NTA affinity chromatography (Qiagen, Valencia, California, USA). Using the imidazoles that gradually increase the concentration, we removed the His-tagged FAM19A5 protein from the Ni-post. Protein expression in the solution is measured using Coomassie Brilliant Blue R-250 dye. Only the solution containing FAM19A5 imidazoles is adopted, and we use PBS to concentrate the FAM19A5 protein. When the concentration is complete, the purity and concentration of the FAM19A5 protein are measured using Western blotting. The concentrated protein is subsequently used to screen the FAM19A5 specific antibody.

[0345] Example 2 Generation of Antibody Library FAM19A5

[0346] 1. Immunization

[0347] Synthesize FAM19A5 peptide, conjugate with KLH (Anygen) at C-terminus, and be used as antigen for chicken immunity.50 μ L of synthetic peptide (VTLDRDSSQPRRTIARQT) KLH conjugate (SEQ ID NO:262) is mixed in 750 μ L of phosphate buffered saline (PBS), and incubate at 37 DEG C for 30 minutes.Then, in 2% squalene endotoxin MPL (monophosphate lipid A species) and mycobacteria (mycobacteria) of the cell wall component of TDW and CWS containing emulsified oil-in-water emulsion adjuvant (RIBI+MPL+TDM+CWS adjuvant, Sigma, St.Louis, Mo, USA), toxin is removed, and then it is subcutaneously injected into three chickens.These chickens are immunized three times in total, and the interval between immunization is about 2-3 weeks.Use the lysate of HEK293T cells overexpressing FAM19A5 protein, the titer of antibody obtained from immunized chicken is measured by immunoblotting. Sera from chickens that received three immunizations were used as primary antibodies. The secondary antibody used was an anti-chicken IgG(Y) polyclonal antibody conjugated to HPR (horseradish peroxidase) (rabbit anti-chicken IgG(Y)-HRP, Millipore corporation, Billerica, MA, USA).

[0348] 2. Preparation of single-chain variable fragment (scFv) library from immunized chickens

[0349] We extracted RNA from the spleen, bone marrow, and synovial bursa of the immunized chickens described above using TRI reagent (Invitrogen, Carlsbad, CA, USA). TM The first strand cDNA is synthesized by using the PCR amplification system (Invitrogen). For the cDNA obtained from the immune system of chicken, the single-stranded variable region library is generated using the Expand High Fidelity PCR System (Roche Molecular Systems, Indiana, USA). In each reaction, 1 μL of cDNA, 60 pmol of each primer, 10 μL of 10 × reaction buffer solution, 8 μL of 2.5 mM dNTP (Promega, Madison, Wisconsin, USA) and 0.5 μL of Taq DNA polymerase are mixed with water. The final volume is 100 μL, and the PCR reaction is carried out under the following conditions: (i) 15 seconds at 94°C, (ii) 30 seconds at 56°C and (iii) 30 cycles of 90 seconds at 72°C, followed by final extension for 10 minutes at 72°C. The PCR product containing a fragment of about 350 bp in length was loaded onto a 1.5% agarose gel, and after electrophoresis, the nucleotide fragments were purified using a QIAGEN Gel II extraction kit (Qiagen, Valencia, CA, USA). The purified PCR product was quantified by reading at OD 260 nm. (1 unit OD = 50 μL / mL).

[0350] The two VH and VL first products from the second PCR are randomly connected by overlap extension PCR. Each PCR reaction is mixed with 100ng of purified VL and VH products, 60pmol of each primer, 10 μL of 10 × reaction buffer, 8 μL of 2.5mM dNTP, 0.5 μL of Taq DNA polymerase and water, and the final volume is 100. PCR is carried out under the following conditions: (i) 15 seconds at 94°C, (ii) 25 cycles of 30 seconds at 56°C and (iii) 2 minutes at 72°C, followed by final extension of 10 minutes at 72°C. The PCR product containing the single-stranded variable region fragment of about 700bp in length is loaded on 1.5% agarose gel, and after electrophoresis, the nucleotide fragment is purified using QIAGEN II gel extraction kit (Qiagen). The PCR product of quantitative purification is read at OD 260nm. (1 unit OD=50 / ml).

[0351] 3. Library, Ligation and Transformation

[0352] The scFv fragment of the PCR product and the vector pComb3X-SS (The Scripps Research Institute, CA, USA) were digested with Sfi I restriction enzyme. 10 μg of the purified overlapping PCT product was mixed with 360 units of Sif I (per 16 units of μg DNA, Roche Molecular Systems, Pleasanton, CA, USA), 20 μL of 10× reaction buffer and water to a final volume of 200 μL. 20 μg of pComb3X-SS vector was mixed with 120 units of Sfi I (per 6 units of μg DNA), 20 μL of 10× reaction buffer and water to a final volume of 200 μL. The mixture was digested at 50°C for 8 hours. Then, the digestion product containing scFv fragment (about 700 bp) and vector (about 3400 bp) was loaded on 1% agarose gel and purified using Gel Extraction Kit II QIAGEN (Qiagen, Valencia, California, USA). 1400 ng of Sfi I restriction enzyme pComb3X vector and 700 ng of digested scFv fragment were mixed with 5× ligase buffer, 10 μL of T4 DNA ligase (Invitrogen, Carlsbad, California, USA) and water to a final volume of 200 μL. The mixture was incubated at 16° C. for 16 hours for ligation.

[0353] After precipitation with ethanol, the DNA precipitate was dissolved in 15 μL of water. In order to produce the library, the connection sample was transformed into Escherichia coli strain ER2738 (New England Biolabs Inc, Hitchin SG4 OTY, Hertfordshire, England, UK) by electroporation using a vibrator gene (Gene Pulser: Bio-Rad Laboratories, Hercules, California, USA). The cells were mixed in 5ml of super broth (SB) culture medium and incubated for one hour at 37°C while stirring at 250rpm. Then, 3 μL of 100mg / mL of kanamycin was added to 10mL of SB culture medium. In order to determine the library size, 0.1 μL, 1 μL and 10 μL of culture samples were applied to Luria broth (LB) agar plates containing 50 μg / ml of kanamycin. After stirring for 1 hour, 4.5 μL of 100mg / mL of kanamycin were added to the LB culture, and further stirred for another 1 hour. Then, 2 ml of VCM13 helper phage (>10 11cfu / ml) together with the preheated LB (183mL) of the 100mg / mL kanamycin containing 92.5 μL are added in LB culture medium.This mixture is stirred for another 2 hours at 37 ℃ with 250rpm.Then, the kanamycin of 280 μL (50mg / mL) is added in the culture, and stirred overnight at 37 ℃.The next day, high-speed centrifuge (Beckman, JA-10 rotor) is used to centrifuge bacteria precipitation at 4 ℃ with 3,000g.Afterwards, the bacteria precipitation is used to extract phagemid DNA, and the supernatant is transferred to a sterile centrifuge bottle simultaneously.Next, 8g of polyethylene glycol-8000 (PEG-8000, Sigma (Sigma)) and 6g of sodium chloride (NaCl, Merck (Merck)) are added in the supernatant, then kept in ice for 30 minutes.Then, the supernatant is centrifuged at 4 ℃ for 15 minutes with 15,000g. The supernatant was then discarded, and the pellet of BSA-propagated phage was suspended in Tris-buffered saline (TBS) containing 1%.

[0354] Example 3: Library panning on immobilized antigen (biopanning)

[0355] Biopanning was performed using magnetic beads (Dynabeads M-270Epoxy, Invitrogen). The beads and proteins were stirred together at room temperature for 20 h while rotating. 7 Each bead is coated with 5 μg of recombinant FAM19A5 protein. Once the coating is completed, the beads are washed 4 times with phosphate buffered saline (PBS) and blocked for one hour at room temperature in PBS containing 3% BSA. Then, the coated beads are cultured at room temperature for two hours with the scFv displayed by the above-mentioned phage. In order to remove any phage that is not combined with the beads coated with antigen, the beads are washed with 0.05% Tween 20 / PBS. Then the bound phage is eluted with 50 μL of 0.1M glycine / hydrochloride (0.1M glycine-HCl, pH 2.2), and neutralized with 3 μL of 2M Tris (tris-HCl, pH 9.1) containing hydrogen chloride. This phage-containing supernatant is used to infect Escherichia coli ER2738 cells, and VCSM13 auxiliary phage is used to amplify and rescue overnight. In addition, by the culture of phage infection on the LB agar plate containing 50 μ g / mL kanamycin trace determination from the phage titer of the culture of phage infection input (input) and output (output).On the second day, PEG-8000 and NaCl were used to precipitate phage, which was then used for bio-panning.By repeating the above process, five bio-pannings were carried out in total.In the case of each amplification, phage was screened and the high affinity to FAM19A5 protein was selected.

[0356] Example 4 Clone selection by phage ELISA

[0357] To analyze the clones selected from the bio-panning, we randomly selected individual clones from the scFv displayed by phage, and used ELISA to confirm that the clones were bound to the FAM19A5 recombinant protein. The FAM19A5 recombinant protein was diluted in 0.1M NaHCO3 buffer, and 100ng / well of protein was used to coat 96-well microtiter plates at 4°C for 16 hours. The next day, the plates were blocked with 3% BSA / PBS at 37°C for 1 hour. Then, the phage supernatant was mixed with 6% BSA / PBS and cultured at 37°C for 2 hours. The plates containing the supernatant were then washed with 0.05% Tween 20 / PBS. The M13 antibody conjugated with HRP (a-M13-HRP, Pierce Chemical Company, Rockford, IL, USA) was diluted to 1 / 5000. 50 μl of the diluted antibody was added to the plate and incubated at 37°C for 1 hour. After incubation and washing, 0.05 M citrate buffer solution, 1 μg / mL of 2,2'-azido-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS, Amresco, Solon, OH, USA) and 0.1% H2O2 were added to the plate for color development. The absorbance of each well was measured at 405 nm. Figure 1A , 1B 1A and 1C show the analysis of 96 clones from level 3, 4 or 5 biopanning of the first, second and third chickens.

[0358] As in Figures 1A to 1C As shown in , we analyzed 24 clones that bound to FAM19A5 recombinant protein and showed high absorbance, and from the 24 clones, we obtained 13 scFv clones with unique sequences. After further selection of clones, we obtained clone 1-65 with the highest affinity.

[0359] Example 5 Generation of anti-FAM19A5-IGG2 / 4 antibodies

[0360] 1. Subcloning of anti-FAM19A5 scFv into mammalian expression vector

[0361] In the FAM19A5 scFv gene sequence, the human Cκ gene is connected to the light chain variable domain, and the human immunoglobulin isotype IgG2 / 4 of the CH1, CH2 and CH3 genes is connected to the heavy chain variable region. Antibodies with each light chain and each heavy chain are synthesized by adding restriction sites (Genscript, USA). The synthesized gene is inserted into a mammalian cell expression vector with modified restriction sites to facilitate cloning. First, the light chain gene is inserted into the vector using Hind III and Xba I (New England Biolabs, UK) restriction enzymes, and then the heavy chain gene is added to the vector by using NheI and BamHI (New England Biolabs, UK) restriction enzymes ( Figure 2 ).

[0362] 2. Purification of anti-FAM19A5 antibodies

[0363] To express and purify anti-FAM19A5-IgG2 / 4 antibodies, we used a mammalian cell transfection and overexpression injection system. We mixed 2 μg / mL of mammalian expression vector with 4 μg of polyethyleneimine (PEI, Polysciences, Warrington, PA, USA) in 150 mM sodium chloride (NaCl, Merck), which was equivalent to 1 / 10 of the cell culture volume. The mixture was allowed to stand at room temperature for 15 minutes. The mixture was added to HEK293F cells (2 × 10 6 cells / ml, Invitrogen) and then in FREESTYLE containing 100U / ml penicillin and streptomycin (Invitrogen). TM 293 expression medium at 7% CO2 and 37 ° C and incubated for six days under stirring conditions of 135 rpm. To purify the expressed anti-FAM19A5 IgG2 / 4 antibody from the cell culture supernatant, we used protein A beads (RepliGen, Waltham, MA, USA) affinity gel chromatography. The protein A chromatography purified antibody was run on 4-12% Bis-Tris gradient gel electrophoresis. The size and yield of the protein were confirmed by Coomassie brilliant blue staining ( Figure 3 ).

[0364] Example 6 Evaluation of mechanical hyperalgesia after in vivo administration of anti-FAM19A5 antibodies in a rat model of chronic constriction injury

[0365] In order to study whether the neutralization of FAM19A5 activity in vivo can relieve neuropathic pain, as Bennett and Xie, Pain 33 (1): 87-107 (1988), Austin et al., J Vis Exp 61: 3393 (2012) previously described, a rat model of chronic contraction injury (CCI) was used. Experimental CCI of the sciatic nerve is one of the most widely used models for studying neuropathic pain, and it is reported that an inflammatory response is induced in the ipsilateral hind paw. Therefore, the hind paw withdrawal threshold measured by, for example, the von Frey test is used as a good indicator of neuropathic pain.

[0366] 1. Inducing neuropathic pain through chronic constriction injury

[0367] Briefly, Zoletil 50 (Vic, France) and xylazine ( Bayer AG (Bayer AG, Germany) IP (intraperitoneal injection) is used to deeply anesthetize 6-week-old male Sprague-Dawley rats. Then, the hair of the lower back and thigh of the rat is shaved, and the skin is disinfected with povidone iodine. Next, the skin of the lateral surface of the thigh is incised, and the sciatic nerve is exposed at the level of the middle thigh by blunt incision via the biceps femoris. At the proximal end of the trifurcation of the sciatic nerve, about 7mm of the nerve has no adherent tissue, and 4 ligatures (4.0 black silk) are tied loosely around it at intervals of about 1mm. The length of the nerve thus affected is 4-5mm long. After performing nerve ligation, the muscle layer and the skin layer are sutured in layers with thread immediately, and local antibiotics are used.

[0368] 2. Administration of anti-FAM19A5 antibodies

[0369] Zoletil 50 (Vic, France) and xylazine ( Male Sprague-Dawley rats were anesthetized with human IgG (Bayer, Germany) and divided into four groups as shown in Table 8. One group of rats with chronic constriction injury ("CCI-induced rats" (G3, n = 10) received anti-FAM19A5 antibody (10 μg / rat in a volume of 0.1 mL) by intrathecal injection. The antibody was administered once a week for 2 weeks (i.e., day 7 and day 14 after CCI induction). Another group of CCI-induced rats (G2, n = 10) was used as a "negative control" and received only human IgG. Another group of CCI-induced rats (G4, n = 10) was treated with pregabalin, a drug used to treat neuropathic pain. The remaining group of rats (G1, n = 10) was used as a "sham control" (i.e., no CCI induction and no drug administration).

[0370] Table 8

[0371]

[0372] 3. Von Frey Test

[0373] On days 6 and 21 after chronic constriction of the sciatic nerve, the paw withdrawal threshold was assessed using the von Frey test. Rats were placed in an apparatus with a metal mesh floor and allowed to stabilize in the environment for approximately 20 minutes. The paw withdrawal threshold was then measured by applying a von Frey filament (0.5 mm in diameter) to the plantar surface of the hind paw three times at 10 second intervals through the mesh floor.

[0374] result

[0375] like Figure 5A and 5B As shown, in vivo administration of anti-FAM19A5 antibodies after CCI induction resulted in a significant increase in paw withdrawal thresholds. Early after administration (e.g., day 6), there were no significant differences between the groups. However, around day 21 after chronic constriction of the sciatic nerve, clear differences between the treated groups (i.e., anti-FAM19A5 antibodies (G3) or pregabalin (G4)) and the negative control group (G2) were significant. For example, on day 6, the mean paw withdrawal thresholds of the negative control (G2) and anti-FAM19A5 antibody treated rats (G3) were 5.1 ± 1.0 and 6.6 ± 2.0, respectively. Figure 5A However, on day 21 after chronic constriction of the sciatic nerve, the mean threshold for the negative control (G2) was 5.5 ± 1.3, while for the anti-FAM19A5 antibody-treated group (G3), the mean threshold was 8.5 ± 1.7 (p < 0.01), which was comparable to the mean threshold for pregabalin (G4: 8.5 ± 1.7), a drug used to treat neuropathic pain ( Figure 5B ). These results suggest that neutralizing FAM19A5 activity by administering anti-FAM19A5 antibodies in vivo can improve neuropathic pain.

[0376] Example 7 Evaluation of motor function after in vivo administration of anti-FAM19A5 antibodies

[0377] In order to study whether anti-FAM19A5 antibody treatment can change motor function, the motor activity of rats induced by CCI was evaluated using the rotarod test. This test is used as a good indicator of any pain or muscle weakness in the lower limbs caused by CCI induction (Chen L et al., 2014, Vadakkan KI et al., 2005). CCI induction and anti-FAM19A5 antibody administration were performed as described in Example 1.

[0378] 1. Rotarod test

[0379] Each Sprague-Dawley rat (sham control and CCI-induced) was carefully placed on a rotarod (Biological Research Apparatus 7750, UGO BASILE, Italy), and the rotation speed of the rotarod was increased from 4 rpm to 20 rpm at regular intervals. The latency to fall was recorded on the 6th day (baseline) and the 21st day after the administration of anti-FAM19A5 antibodies. Motor performance was considered to be the latency to fall from the rotarod device, which was determined based on the average time of three trials per rat.

[0380] 2. Results

[0381] like Fig. 6A and 6B As shown, in vivo administration of anti-FAM19A5 antibody after CCI induction resulted in improved latency times (i.e., rats stayed on the rotarod apparatus longer). Although there were no significant differences between the groups on day 6 after chronic constriction of the sciatic nerve ( Fig. 6A ), but by day 21, anti-FAM19A5 antibody-treated rats showed a much longer latency compared with the negative control (G2: 64.3 ± 14.1 vs. G3: 92.8 ± 11.5), which was comparable to the latency of pregabalin (G4: 91.1 ± 17.8) ( Figure 6B ). These results suggest that in vivo administration of anti-FAM19A5 antibodies after CCI induction can not only alleviate neuropathic pain but also improve motor function.

[0382] Example 8: Evaluation of mechanical hyperalgesia following in vivo administration of anti-FAM19A5 antibodies in a rat model of diabetic peripheral neuropathy

[0383] In order to further evaluate the benefits of neutralizing FAM19A5 activity to neuropathic pain, a rat model of diabetic peripheral neuropathy was used. By administering streptozotocin (STZ) (50mg / kg, intraperitoneal) to animals, diabetic peripheral neuropathy (DPN) was induced in Sprague-Dawley rats. Blood glucose levels were measured about one week after STZ administration, and only animals with fasting blood glucose levels of 300mg / dL or higher were selected for antibody administration. About 3 weeks after STZ administration, selected DPN rats received control antibodies (normal human IgG, "NHI") or anti-FAM19A5 antibodies (3-2) for weekly administration (a total of 8 administrations, i.e., from the 4th week to the 11th week after STZ administration). Antibodies were administered intrathecally to animals at a dosage of 25g / rats. Initial (healthy) animals were used as "sham controls" (i.e., no DPN induction and no antibody administration). Paw withdrawal thresholds were measured in animals using the von Frey test as described in Example 6 at 7 and 11 weeks after STZ administration (ie, 4 and 8 weeks, respectively, after the start of antibody administration).

[0384] like Fig.10 As shown, DPN animals receiving anti-FAM19A5 antibodies had significantly higher paw withdrawal thresholds than those of DPN animals receiving control antibodies. This increase in thresholds was observed at both the 7th and 11th weeks after STZ administration (i.e., the 4th and 8th weeks after antibody administration, respectively).

[0385] Example 9: Evaluation of thermal hyperalgesia following in vivo administration of anti-FAM19A5 antibodies in a rat model of diabetic peripheral neuropathy

[0386] Next, to evaluate the effect of anti-FAM19A5 administration on neuropathic pain associated with thermal hyperalgesia, diabetic peripheral neuropathy (DPN) was induced in Sprague-Dawley rats, and control (NHI) or anti-FAM19A5 antibody (3-2) was administered to the animals as described above in Example 8. Then, at weeks 4 and 8 after the start of antibody administration (i.e., weeks 7 and 11 after STZ administration), the animals were placed on a hot plate and the temperature was immediately raised to 55° C. Then, the time it took for the animals to respond to the increased temperature (e.g., by jumping or licking their paws) was measured.

[0387] like Fig.11A and 11BAs shown, DPN animals receiving control antibodies were much more sensitive to thermal stimulation, with a paw withdrawal latency of approximately 6 seconds. In contrast, DPN rats receiving anti-FAM19A5 antibodies had significantly higher paw withdrawal latency times at both weeks 4 and 8. The above data further confirm that neutralizing FAM19A5 activity can improve neuropathic pain associated with mechanical and thermal hyperalgesia.

[0388] Example 10 Analysis of sensory nerve conduction velocity after in vivo administration of anti-FAM19A5 antibodies in a rat model of diabetic peripheral neuropathy

[0389] In order to determine the impact of anti-FAM19A5 administration on peripheral sensory nerve damage and / or dysfunction, sensory nerve conduction velocity (SNCV) test was performed. By measuring the speed at which electrical signals are transmitted from descending sensory nerves to ascending sensory nerves, this test can help identify potential nerve damage and / or dysfunction. As described in Example 8, diabetic peripheral neuropathy was induced in Sprague-Dawley rats using STZ, and controls or anti-FAM19A5 antibodies (3-2) were administered to animals. Then, 8 weeks after the start of antibody administration, the nerve conduction velocity in the sural nerve was measured. Viking Quest (Natus Neurology Incorporated, U.S.) equipment was used to induce electrical stimulation at 10 mA and 10 μV.

[0390] like Fig.12 As shown, DPN rats receiving anti-FAM19A5 antibodies had improved conduction velocity compared to those DPN animals receiving control antibodies. This result suggests that neutralizing the FAM19A5 activity can improve and / or reduce peripheral nerve damage / dysfunction, thereby helping to alleviate neuropathic pain, as observed in the previous examples. Overall, the above results indicate that FAM19A5 antagonists (such as the anti-FAM19A5 antibodies disclosed herein) may be a suitable treatment option for neuropathic pain.

[0391] Example 11 Epitope Mapping

[0392] Overlapping peptide fragments of human FAM19A5 protein (F1-F5, see Fig. 9) and conjugated with BSA. The binding of monoclonal antibody 1-65 to BSA-conjugated peptide fragments F1-F6 was determined by Western blot analysis. For Western blot analysis, BSA-conjugated FAM19A5 fragments F1-F6 were separated by SDS polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes by standard procedures. The membrane was incubated with anti-FAM19A5 antibody 1-65 (2 μg / ml, 1-65-ScFv-rabbit Fc-SSS), and the antigen-antibody complex was detected with a suitable secondary antibody conjugated with horseradish peroxidase (anti-rabbit IgG (Fc specific)-HRP, 1:4000 dilution). Fig. 9 ).like Fig. 9 As shown, anti-FAM19A5 antibody 1-65 strongly binds to fragment F5. In addition to F5, it also binds to fragments F2 and F3.

[0393] It should be understood that the Detailed Description section, rather than the Summary and Abstract sections, is intended to be used to interpret the claims. The Summary and Abstract sections may set forth one or more but not all exemplary embodiments of the present disclosure as contemplated by the inventors, and therefore, are not intended to limit the present disclosure and the appended claims in any way.

[0394] The present disclosure has been described with the aid of functional building blocks that illustrate the implementation of specified functions and their relationships. For ease of description, the boundaries of these functional building blocks have been arbitrarily defined in this article. Alternative boundaries can be defined as long as the specified functions and their relationships are appropriately performed.

[0395] The foregoing description of specific embodiments will reveal the general nature of the present disclosure so fully that others can easily modify and / or adapt various applications, such as specific embodiments, without departing from the general concepts of the present disclosure by applying knowledge within the art without undue experimentation. Therefore, based on the teachings and guidance given herein, such adaptations and modifications are intended to be within the meaning and range of equivalents of the disclosed embodiments. It should be understood that the wording or terminology herein is for descriptive purposes and not for limitation, so that the terms or wording of this specification will be interpreted by the technician based on the teachings and guidance.

[0396] The breadth and scope of the present disclosure should not be limited by any of the above-described exemplary embodiments, but should be defined only in accordance with the following claims and their equivalents.

[0397] All publications, patents, patent applications, internet sites, and accession / database sequences (including 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 / database sequence was specifically and individually indicated to be incorporated by reference.

[0398] This PCT application claims the benefit of priority to U.S. Provisional Application No. 62 / 661,923, filed on April 24, 2018, which is incorporated herein by reference in its entirety.

Claims

1. An antagonist to family similarity 19 member A5 (FAM19A5) protein ("FAM19A5 antagonist") for use in treating neuropathic pain in a subject in need thereof. 2 . The FAM19A5 antagonist for use according to claim 1 , wherein the neuropathic pain is central neuropathic pain or peripheral neuropathic pain.

3. The FAM19A5 antagonist for use according to claim 1 or 2, wherein the neuropathic pain is associated with physical injury, infection, diabetes, cancer therapy, alcoholism, amputation, muscle weakness in the back, legs, buttocks or face, trigeminal neuralgia, multiple sclerosis, herpes zoster, spinal surgery or any combination thereof.

4. The FAM19A5 antagonist for use according to any one of claims 1 to 3, wherein the neuropathic pain comprises carpal tunnel syndrome, central pain syndrome, degenerative disc disease, diabetic neuropathy, phantom limb pain, postherpetic neuralgia (shingles), pudendal neuralgia, sciatica, low back pain, trigeminal neuralgia, or any combination thereof. 5 . The FAM19A5 antagonist for use according to any one of claims 1 to 4 , wherein the neuropathic pain is caused by compression of a nerve.

6. The FAM19A5 antagonist for use according to claim 4, wherein the diabetic neuropathy is diabetic peripheral neuropathy.

7. The FAM19A5 antagonist for use according to claim 4 or 5, wherein the neuropathic pain is sciatica.

8. A FAM19A5 antagonist for use in increasing the threshold or latency to an external stimulus in a subject in need thereof. 9 . The FAM19A5 antagonist for use according to claim 8 , wherein the external stimulus is a mechanical stimulus. 10 . The FAM19A5 antagonist for use according to claim 8 , wherein the external stimulus is thermal stimulation.

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