Human antibodies against artemin and methods of use thereof

Abstract

The present disclosure provides antibodies and antigen-binding fragments thereof that bind to human artemin. Also provided are methods of using anti-artemin antibodies and antigen-binding fragments.

Description

Technical Field

[0001] The present disclosure relates to human antibodies and antigen-binding fragments of human antibodies that specifically bind artemin, and methods of using the same.

[0002] Sequence Listing

[0003] A formal copy of the Sequence Listing is submitted electronically via EFS-Web simultaneously with the specification as an ASCII-formatted sequence listing entitled 10907WO01_Sequence_Listing_ST25, created on April 19, 2022, and having a size of approximately 69,632 bytes. The sequence listing contained in this ASCII-formatted document is part of the specification and is hereby incorporated by reference in its entirety. Background Art

[0004] Artemin (ARTN) is a neurotrophic factor that is a member of the glial cell line-derived neurotrophic factor (GDNF) ligand family. Each GDNF family ligand binds to a glycosylphosphatidylinositol (GPI)-anchored receptor associated with the plasma membrane. This receptor family is called the GDNF family receptor (GFRα) family and is composed of four different receptors, GFRα1-4. ARTN preferentially binds to the receptor GFRα3 and is the only known ligand for GFRα3. ARTN and other GDNF family ligands signal through the RET ("rearranged during transfection") receptor tyrosine kinase, which was originally discovered as an oncogene. RET is activated by GDNF family members only when the ligand first binds to its GFRα receptor (Airaksinen et al., Nature Reviews Neuroscience (2002), 3:383-394).

[0005] ARTN and GFRα3 are highly expressed during development and are involved in the development of the sympathetic nervous system. In adult mice, ARTN is expressed in the testis, uterus, thyroid, prostate, and epididymis, as well as in small arteries in the olfactory bulb and intestine and mesentery (Airaksinen et al., Nature Reviews Neuroscience (2002), 3:383 - 394; Airaksinen et al., Brain, Behavior and Evolution, (2006), 68:181 - 190). GFRα3 expression in adults is mainly limited to sensory neurons of the dorsal root ganglia (DRG) (Orozco et al., European J. Neuroscience, (2001), 13:2177 - 2182). Mice lacking ARTN or GFRα3 exhibit abnormalities in the migration and axonal projection patterns of the sympathetic nervous system, resulting in abnormal innervation of target tissues (Onma et al., Neuron, (2002), 35:267 - 282).

[0006] The potential roles of ARTN and GFRα3 in pain and heat perception have been shown in several studies. For example, it has been demonstrated that injecting artemin protein into the hindpaw of rodents leads to thermal hyperalgesia, and this nociception is enhanced when artemin is co - injected with NGF (Malin et al., J. Neuroscience, (2006), 26(33):8588 - 8599). Other studies have shown that artemin mRNA expression is upregulated in murine inflammatory models, and artemin transgenic mice have high expression of TRPV1 and TRPA1 and high behavioral sensitivity to heat and cold (Elitt et al., J. Neuroscience, (2006), 26(33):8578 - 8587). In addition, artemin and GFRα3 expression is increased in a rat model of migraine (Shang et al., JHeadache Pain, (2016), 17(1):81) and in dogs treated with radiotherapy (Nolan et al., RadiatRes, (2020), 193(3):241 - 248). Artemin expression is also increased in the tongue mucosa of patients with burning mouth syndrome and in a mouse model of burning mouth syndrome (Shinoda et al., PAIN, (2015), 156(12):2528 - 2537). Summary of the Invention

[0007] In one aspect, there is provided an isolated antibody or an antigen-binding fragment thereof that specifically binds artemin. In some embodiments, the antibody is a monoclonal antibody that binds human artemin or an antigen-binding fragment thereof. In some embodiments, the antibody or its antigen-binding fragment has a K as measured by surface plasmon resonance at 25°C of less than 5 nM D that binds to human artemin.

[0008] In some embodiments, the antibody or its antigen-binding fragment:

[0009] (a) blocks the binding of at least 65% of human artemin to a solid support coated with human GFRα3, as measured by ELISA;

[0010] (b) inhibits or reduces artemin-activated signal transduction through the GFRα3 and RET receptor tyrosine kinases; and / or

[0011] (c) inhibits or reduces artemin-induced thermal hyperalgesia in vivo.

[0012] In some embodiments, the antibody or its antigen-binding fragment cross-reacts with artemin from at least one non-human species. In some embodiments, the antibody or its antigen-binding fragment cross-reacts with monkey artemin. In some embodiments, the antibody or its antigen-binding fragment cross-reacts with mouse artemin.

[0013] In some embodiments, the antibody or its antigen-binding fragment comprises:

[0014] (a) a heavy chain complementarity determining region 1 (HCDR1) comprising an amino acid sequence selected from the group consisting of SEQ ID NO:4, 24, 44, and 64;

[0015] (b) a heavy chain complementarity determining region 2 (HCDR2) comprising an amino acid sequence selected from the group consisting of SEQ ID NO:6, 26, 46, and 66;

[0016] (c) a heavy chain complementarity determining region 3 (HCDR3) comprising an amino acid sequence selected from the group consisting of SEQ ID NO:8, 28, 48, and 68;

[0017] (d) a light chain complementarity determining region 1 (LCDR1) comprising an amino acid sequence selected from the group consisting of SEQ ID NO:12, 32, 52, and 72;

[0018] (e) a light chain complementarity determining region 2 (LCDR2) comprising an amino acid sequence selected from the group consisting of SEQ ID NO:14, 34, 54, and 74; and

[0019] (f) A light chain complementarity determining region 3 (LCDR3) comprising an amino acid sequence selected from the group consisting of SEQ ID NO:16, 36, 56, and 76.

[0020] In some embodiments, the antibody or antigen-binding fragment thereof comprises:

[0021] (a) An HCDR1 comprising the amino acid sequence of SEQ ID NO:4, an HCDR2 comprising the amino acid sequence of SEQ ID NO:6, an HCDR3 comprising the amino acid sequence of SEQ ID NO:8, an LCDR1 comprising the amino acid sequence of SEQ ID NO:12, an LCDR2 comprising the amino acid sequence of SEQ ID NO:14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:16; or

[0022] (b) An HCDR1 comprising the amino acid sequence of SEQ ID NO:24, an HCDR2 comprising the amino acid sequence of SEQ ID NO:26, an HCDR3 comprising the amino acid sequence of SEQ ID NO:28, an LCDR1 comprising the amino acid sequence of SEQ ID NO:32, an LCDR2 comprising the amino acid sequence of SEQ ID NO:34, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:36; or

[0023] (c) An HCDR1 comprising the amino acid sequence of SEQ ID NO:44, an HCDR2 comprising the amino acid sequence of SEQ ID NO:46, an HCDR3 comprising the amino acid sequence of SEQ ID NO:48, an LCDR1 comprising the amino acid sequence of SEQ ID NO:52, an LCDR2 comprising the amino acid sequence of SEQ ID NO:54, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:56; or

[0024] (d) An HCDR1 comprising the amino acid sequence of SEQ ID NO:64, an HCDR2 comprising the amino acid sequence of SEQ ID NO:66, an HCDR3 comprising the amino acid sequence of SEQ ID NO:68, an LCDR1 comprising the amino acid sequence of SEQ ID NO:72, an LCDR2 comprising the amino acid sequence of SEQ ID NO:74, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:76.

[0025] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (HCVR) comprising an amino acid sequence having at least 85% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:2, 22, 42, and 62.

[0026] In some embodiments, the antibody or antigen-binding fragment thereof comprises a light chain variable region (LCVR) comprising an amino acid sequence having at least 85% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 10, 30, 50, and 70.

[0027] In some embodiments, the antibody or antigen-binding fragment thereof comprises:

[0028] (a) an HCVR comprising the amino acid sequence of SEQ ID NO:2 and an LCVR comprising the amino acid sequence of SEQ ID NO:10; or

[0029] (b) an HCVR comprising the amino acid sequence of SEQ ID NO:22 and an LCVR comprising the amino acid sequence of SEQ ID NO:30; or

[0030] (c) an HCVR comprising the amino acid sequence of SEQ ID NO:42 and an LCVR comprising the amino acid sequence of SEQ ID NO:50; or

[0031] (d) an HCVR comprising the amino acid sequence of SEQ ID NO:62 and an LCVR comprising the amino acid sequence of SEQ ID NO:70.

[0032] In some embodiments, the antibody or antigen-binding fragment thereof comprises:

[0033] (a) a heavy chain comprising an amino acid sequence having at least 85% sequence identity to the amino acid sequence of SEQ ID NO:18, and a light chain comprising an amino acid sequence having at least 85% sequence identity to the amino acid sequence of SEQ ID NO:20; or

[0034] (b) a heavy chain comprising an amino acid sequence having at least 85% sequence identity to the amino acid sequence of SEQ ID NO:38, and a light chain comprising an amino acid sequence having at least 85% sequence identity to the amino acid sequence of SEQ ID NO:40; or

[0035] (c) a heavy chain comprising an amino acid sequence having at least 85% sequence identity to the amino acid sequence of SEQ ID NO:58, and a light chain comprising an amino acid sequence having at least 85% sequence identity to the amino acid sequence of SEQ ID NO:60; or

[0036] (d) A heavy chain comprising an amino acid sequence having at least 85% sequence identity to the amino acid sequence of SEQ ID NO:78, and a light chain comprising an amino acid sequence having at least 85% sequence identity to the amino acid sequence of SEQ ID NO:80.

[0037] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO:18 and a light chain comprising the amino acid sequence of SEQ ID NO:20. In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO:38 and a light chain comprising the amino acid sequence of SEQ ID NO:40. In some embodiments, the antibody or antigen-binding fragment thereof comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO:58 and a light chain comprising the amino acid sequence of SEQ ID NO:60. In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:78 and a light chain comprising the amino acid sequence of SEQ ID NO:80.

[0038] In some embodiments, the antibody or antigen-binding fragment thereof is fully human.

[0039] In another aspect, there is provided a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof and a pharmaceutically acceptable carrier.

[0040] In another aspect, there is provided a nucleic acid molecule comprising a nucleotide sequence encoding the antibody or antigen-binding fragment thereof. In some embodiments, the nucleic acid molecule comprises one or more nucleotide sequences listed in Table 9.

[0041] In yet another aspect, there are provided an expression vector and a host cell comprising the nucleic acid molecule disclosed herein, and a method for producing an anti-artemin antibody using the nucleic acid molecule, expression vector and host cell disclosed herein.

[0042] In another aspect, there is provided a method for inhibiting the signal transduction of artemin activation in cells. In some embodiments, the method comprises contacting the cell with the antibody or antigen-binding fragment or pharmaceutical composition disclosed herein, thereby inhibiting the signal transduction of artemin activation in the cell. In some embodiments, the cell is in a subject. In some embodiments, the cell is in vitro or ex vivo.

[0043] In another aspect, methods for treating diseases, disorders or conditions associated with artemin expression or activity are provided. In some embodiments, the methods comprise administering an antibody or an antigen-binding fragment thereof or a pharmaceutical composition disclosed herein to a subject in need thereof (e.g., a subject suffering from a disease, disorder or condition associated with artemin expression or activity). In some embodiments, the antibody or an antigen-binding fragment thereof or the pharmaceutical composition is administered in combination with a second therapeutic agent to the subject.

[0044] In another aspect, an anti-artemin antibody or an antigen-binding fragment thereof, or a pharmaceutical composition comprising an anti-artemin antibody or an antigen-binding fragment thereof, for use in a method for treating diseases, disorders or conditions associated with artemin expression or activity is provided. In some embodiments, the methods comprise administering an antibody or an antigen-binding fragment thereof or a pharmaceutical composition disclosed herein to a subject in need thereof (e.g., a subject suffering from a disease, disorder or condition associated with artemin expression or activity). In some aspects, the antibody or an antigen-binding fragment thereof or the pharmaceutical composition is therapeutically combined with a second therapeutic agent. In some aspects, the antibody or an antigen-binding fragment thereof or the pharmaceutical composition is combined with a second therapeutic agent.

[0045] In another aspect, the use of an anti-artemin antibody or an antigen-binding fragment thereof or a pharmaceutical composition comprising an anti-artemin antibody or an antigen-binding fragment thereof in the manufacture of a medicament for use in a method for treating diseases, disorders or conditions associated with artemin expression or activity is provided. In some embodiments, the methods comprise administering an antibody or an antigen-binding fragment thereof or a pharmaceutical composition disclosed herein to a subject in need thereof (e.g., a subject suffering from a disease, disorder or condition associated with artemin expression or activity). In some aspects, the antibody or an antigen-binding fragment thereof or the pharmaceutical composition is therapeutically combined with a second therapeutic agent. In some aspects, the antibody or an antigen-binding fragment thereof or the pharmaceutical composition is combined with a second therapeutic agent.

[0046] In some embodiments, the disease, disorder or condition is acute pain, chronic pain, neuropathic pain, inflammatory pain, trigeminal neuralgia, postherpetic neuralgia, general neuralgia, visceral pain, osteoarthritis pain, gout, radicular pain, sciatica, back pain, head or neck pain, breakthrough pain, postoperative pain, chemotherapy-induced neuropathic pain, radiotherapy-induced neuropathic pain, radiation-related pain or cancer pain. In some embodiments, the disease, disorder or condition is migraine, cluster headache, chronic headache or tension headache.

[0047] Other embodiments will be apparent from a reading of the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 . Binding of different concentrations of hArtemin-MMH (human Artemin) to the surface of an Octet biosensor captured by Artemin mAb.

[0049] Figure 2 . Binding of different concentrations of MfArtemin-MMH (cynomolgus monkey Artemin) to the surface of an Octet biosensor captured by Artemin mAb.

[0050] Figure 3 . Binding of different concentrations of mArtemin-MMH (mouse Artemin) to the surface of an Octet biosensor captured by Artemin mAb.

[0051] Figure 4 . Inhibition of artemin-induced plantar thermal hyperalgesia in animals administered with anti-artemin antibodies (H4H33331P, H4H33335P, H4H33336P or H4H33349P) or isotype control antibodies (negative control) at 10 mg / kg. Detailed Description

[0052] Definitions

[0053] Before describing the present invention, it should be understood that the present invention is not limited to the specific methods and experimental conditions described, as such methods and conditions may vary. It should also be understood that the terms used herein are for the purpose of describing specific embodiments only and are not intended to be limiting, as the scope of the present invention will be limited only by the appended claims.

[0054] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0055] As used herein, when referring to a specifically recited numerical value, the term "about" means that the value may vary from the recited value by no more than 1%. For example, as used herein, the expression "about 100" includes 99 and 101 and all values therebetween (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0056] As used herein, the term "artemin" or "ARTN" refers to a glial cell line-derived neurotrophic factor (GDNF) family ligand. See, Baloh et al., Neuron, (1998), 21(6):1291-1302. As used herein, the term "artemin" can refer to the precursor form or the mature form (i.e., the processed form), such as the full-length processed form or a fragment thereof. In some embodiments, the term "artemin" refers to the human artemin protein or a fragment thereof. In some embodiments, the artemin protein comprises the amino acid sequence of human artemin shown in NCBI accession number NP_476432.2, or a fragment thereof (e.g., a proteolytically processed portion). In some embodiments, the artemin protein comprises the amino acid sequence of human artemin shown in Uniprot accession number Q5T4W7, or a fragment thereof (e.g., a proteolytically processed portion), or an isoform thereof. In some embodiments, the artemin protein comprises the amino acid sequence of the artemin protein or fragment shown in U.S. Patent No. 6,284,540.

[0057] The term "antibody that binds artemin" or "anti-artemin antibody" includes antibodies that specifically recognize monomeric artemin and antigen-binding fragments thereof, as well as antibodies that specifically recognize dimeric artemin and antigen-binding fragments thereof.

[0058] As used herein, the term "antibody" refers to an antigen-binding molecule or molecular complex that comprises a set of complementarity-determining regions (CDRs) that specifically bind to or interact with a particular antigen (e.g., ARTN). As used herein, the term "antibody" includes immunoglobulin molecules that comprise four polypeptide chains (two heavy (H) chains and two light (L) chains) interconnected by disulfide bonds and multimers thereof (e.g., IgM). In a typical antibody, each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or V H ) and a heavy chain constant region. The heavy chain constant region comprises three domains: C H 1, C H 2, and C H 3. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or V L ) and a light chain constant region. The light chain constant region comprises one domain (C L 1). The V H region and the V L region can be further subdivided into hypervariable regions known as complementarity-determining regions (CDRs), which are interspersed with more conserved regions known as framework regions (FRs). Each V H and V LComposed 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, FR4. In some embodiments, the FRs of the antibody (or its antigen-binding portion) may be identical to the human germline sequence, or may be natural or artificially modified. Amino acid consensus sequences can be defined based on the juxtaposition analysis of two or more CDRs.

[0059] As used herein, the term "antibody" also includes antigen-binding fragments of whole antibody molecules. The terms "antigen-binding portion" of an antibody, "antigen-binding fragment" of an antibody, "antigen-binding domain", etc. as used herein include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds an antigen to form a complex. Antigen-binding fragments of an antibody can be derived from whole antibody molecules, for example, using any suitable standard techniques such as proteolytic digestion or recombinant genetic engineering techniques involving the manipulation and expression of DNA encoding the variable domains of the antibody and optionally the constant domains of the antibody. Such DNA is known and / or can be readily obtained, for example, from commercial sources, DNA libraries (including, for example, phage-antibody libraries), or can be synthesized. The DNA can be sequenced and manipulated chemically or by using molecular biology techniques, for example, to arrange one or more variable domains and / or constant domains in a suitable configuration, or to introduce codons, generate cysteine residues, modify, add, or delete amino acids, etc.

[0060] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units composed of amino acid residues that mimic the hypervariable regions of an antibody (e.g., isolated complementarity-determining regions (CDRs), such as CDR3 peptides) or restricted FR3-CDR3-FR4 peptides. Other engineered molecules, such as domain-specific antibodies, single-domain antibodies, domain-deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetra-bodies, microantibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains are also encompassed within the expression "antigen-binding fragment" as used herein.

[0061] Antigen-binding fragments of an antibody will generally include at least one variable domain. The variable domain can have any size or amino acid composition and will generally include at least one CDR adjacent to or in-frame with one or more framework sequences. In antigen-binding fragments having a V H domain associated with a V L domain, the VH and V L domains can be positioned relative to each other in any suitable arrangement. For example, the variable regions can be dimers and contain V H -V H 、V H -V L or V L -V L dimers. Alternatively, the antigen-binding fragment of an antibody can contain a monomeric V H or V L domain.

[0062] In certain embodiments, the antigen-binding fragment of an antibody can contain at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains that can be found within the antigen-binding fragment of an antibody include: (i) V H -C H 1; (ii) V H -C H 2; (iii) V H -C H 3; (iv) V H -C H 1-C H 2; (v) V H -C H 1-C H 2-C H 3; (vi) V H -C H 2-C H 3; (vii) V H -C L ; (viii) V L -C H 1; (ix) V L -C H 2; (x) V L -C H 3; (xi) V L -C H 1-C H 2; (xii) V L -C H 1-C H 2-C H 3; (xiii) V L -C H 2-C H 3; and (xiv) V L -C L。In any configuration of the variable and constant domains (including any of the exemplary configurations listed above), the variable and constant domains can be directly linked to each other or can be connected by a full-length or partial hinge or linker region. The hinge region can consist of at least 2 (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids that create a flexible or semi-flexible linkage between adjacent variable and / or constant domains in a single polypeptide molecule. Additionally, an antigen-binding fragment of an antibody can comprise homodimers or heterodimers (or other multimers) of any of the variable and constant domain configurations listed above that non-covalently bind to each other and / or non-covalently associate with one or more monomeric V H or V L domains (e.g., via disulfide bonds).

[0063] The term "antibody" as used herein also encompasses multispecific (e.g., bispecific) antibodies. Multispecific antibodies or antigen-binding fragments of antibodies generally comprise at least two different variable domains, where each variable domain is capable of specifically binding to a separate antigen or a different epitope on the same antigen. Using conventional techniques available in the art, any form of multispecific antibody can be adapted to the context of the antibodies or antigen-binding fragments of the present disclosure. For example, the present disclosure includes methods that employ bispecific antibodies, where one arm of the immunoglobulin is specific for ARTN or a fragment thereof, and the other arm of the immunoglobulin is specific for a second therapeutic target or conjugated to a therapeutic moiety. Exemplary bispecific forms that can be used in the context of the present disclosure include, but are not limited to, for example, scFv-based or diabody bispecific forms, IgG-scFv fusions, dual variable domain (DVD)-Ig, tetrabody, knob-into-hole, common light chain (e.g., common light chain with knob-into-hole, etc.), CrossMab, CrossFab, (SEED)body, leucine zipper, Duobody, IgG1 / IgG2, dual action Fab (DAF)-IgG, and Mab 2 bispecific forms (for a review of the foregoing forms, see, e.g., Klein et al., 2012, mAbs 4:6, 1-11, and references cited therein). Bispecific antibodies can also be constructed using peptide / nucleic acid conjugates, e.g., where unnatural amino acids with orthogonal chemical reactivity are used to generate site-specific antibody-oligonucleotide conjugates, which are then self-assembled into multimeric complexes with defined composition, valency, and geometry. (See, e.g., Kazane et al., J. Am. Chem. Soc. [e-Ed.: December 4, 2012]).

[0064] As used herein, the term "human antibody" is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Notwithstanding this, a human antibody of the present disclosure may contain amino acid residues that are not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by in vitro random or site-specific mutagenesis or by in vivo somatic mutation), such as in the CDRs, and specifically, in CDR3. However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.

[0065] As used herein, the term "recombinant antibody" is intended to include all antibodies prepared, expressed, produced, or isolated by recombinant means. The term includes, but is not limited to, antibodies expressed using recombinant expression vectors transfected into host cells (e.g., Chinese hamster ovary (CHO) cells, NS0 cells, BHK cells, or HEK293 cells) or cell expression systems; antibodies isolated from recombinant, combinatorial human antibody libraries; and antibodies isolated from non-human animals (e.g., mice, such as human immunoglobulin gene transgenic mice) (see, e.g., Taylor et al. (1992) Nucl. Acids Res. 20:6287-6295). In some embodiments, the recombinant antibody is a recombinant human antibody. In some embodiments, the recombinant human antibody has variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or in vivo somatic mutagenesis when using transgenic animals with human Ig sequences), and thus the amino acid sequences of the V H and V L regions are sequences that, although derived from and related to human germline V H and V L sequences, may not naturally occur within the human antibody germline repertoire in vivo.

[0066] An "isolated antibody" refers to an antibody that has been identified and separated and / or recovered from at least one component of its natural environment. For example, an antibody that has been separated or removed from at least one component of an organism, or from a tissue or cell in which the antibody naturally occurs or is naturally produced, is an "isolated antibody". An isolated antibody also includes an antibody in situ within a recombinant cell. An isolated antibody is an antibody that has undergone at least one purification or isolation step. According to certain embodiments, an isolated antibody may be substantially free of other cellular material and / or chemicals.

[0067] The term "specifically binds", etc., means that an antibody or its antigen-binding fragment forms a relatively stable complex with an antigen under physiological conditions. Specific binding can be characterized by an equilibrium dissociation constant of at least about 1×10 -6 M or less, such as 10-7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M or 10 -12 M (smaller K D indicates a tighter binding). Methods for determining whether an antibody specifically binds an antigen are known in the art and include, for example, equilibrium dialysis, surface plasmon resonance (e.g., BIACORE TM ), biolayer interferometry (e.g., HTX biosensor), solution affinity ELISA, etc. In some embodiments, in surface plasmon resonance assays, specific binding is measured, for example, at 25 °C or 37 °C. An antibody or antigen-binding fragment that specifically binds an antigen from one species may or may not cross-react with other antigens, such as orthologous antigens from another species.

[0068] As used herein, the term "K D " refers to the equilibrium dissociation constant of a particular antibody-antigen interaction.

[0069] As used herein, the term "surface plasmon resonance" refers to an optical phenomenon that allows the analysis of real-time biomolecular interactions by detecting changes in the protein concentration in a biosensor matrix, e.g., using BIACORE TM (Cytiva, Marlborough, MA).

[0070] As used herein, the term "biolayer interferometry" or "BLI" refers to an optical technique used to measure real-time biomolecular interactions by analyzing the interference pattern caused by the binding of a biomolecule dispensed in a sample plate to a second biomolecule immobilized on a biosensor plate, e.g., using system (Sartorius AG, Germany).

[0071] As used herein, the term "epitope" refers to an antigenic determinant that interacts with a specific antigen-binding site (termed a paratope) in the variable region of an antibody molecule. A single antigen can have more than one epitope. Thus, different antibodies can bind to different regions on an antigen and can have different biological effects. The term "epitope" also refers to the site on an antigen to which a B cell and / or T cell responds. It also refers to the region of an antigen to which an antibody binds. Epitopes can be linear or discontinuous (e.g., conformational). Linear epitopes are generated by adjacent amino acid residues in a polypeptide chain. Conformational epitopes are generated by the spatial juxtaposition of amino acids from different segments of a linear polypeptide chain. In certain embodiments, an epitope can comprise determinants that are the chemically reactive surface groups of a molecule, such groups as amino acids, sugar side chains, phosphoryl or sulfonyl groups, and in certain embodiments, can have specific three-dimensional structural features and / or charge-to-mass ratio features. Epitopes can also be defined as structural or functional. Functional epitopes are generally a subset of structural epitopes and have those residues that directly contribute to the affinity of the interaction. Epitopes typically comprise at least 3, and more usually at least 5 or at least 8-10 amino acids in a unique spatial conformation.

[0072] Methods for determining the epitope of an antigen-binding protein (e.g., an antibody or antigen-binding fragment) include alanine-scanning mutagenesis, peptide blotting (Reineke, Methods Mol Biol 2004, 248:443-463), peptide cleavage analysis, crystallographic studies, and NMR analysis. Additionally, methods such as epitope exclusion, epitope extraction, and chemical modification of the antigen can be employed (Tomer, Prot Sci 2000, 9:487-496). Another method for identifying the amino acids in a polypeptide that interact with an antigen-binding protein (e.g., an antibody or antigen-binding fragment) is hydrogen / deuterium exchange detected by mass spectrometry (HDX). See, e.g., Analytical Biochemistry 1999, 267:252-259; Engen and Smith, Anal Chem 2001, 73:256A-265A.

[0073] When used with reference to a nucleic acid or a fragment thereof, the terms "substantially identical" and "substantially the same" indicate that, when optimally aligned with another nucleic acid (or its complementary strand) with appropriate nucleotide insertions or deletions, there is nucleotide sequence identity of at least about 85%, e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, as measured by any well-known sequence identity algorithm discussed herein (such as FASTA, BLAST or GAP). In some cases, a nucleic acid molecule that is substantially identical to a reference nucleic acid molecule may encode a polypeptide having the same or substantially similar amino acid sequence as the polypeptide encoded by the reference nucleic acid molecule.

[0074] When applied to polypeptides, the terms "substantially identical" and "substantially the same" mean that, when optimally aligned, two peptide sequences share at least about 85% sequence identity, e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity. In some embodiments, the non-identical residue positions differ by conservative amino acid substitutions. A "conservative amino acid substitution" is a substitution in which one amino acid residue is replaced with another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). Generally, conservative amino acid substitutions do not substantially alter the functional properties of the protein.

[0075] Sequence analysis software is typically used to measure sequence similarity of polypeptides. Protein analysis software uses similarity metrics assigned to various substitutions, deletions and other modifications that include conservative amino acid substitutions to match similar sequences. For example, GCG software contains programs such as GAP and BESTFIT, which can be used with default parameters to determine sequence homology or sequence identity between closely related polypeptides (e.g., homologous polypeptides from different biological species) or between a wild-type protein and its mutant proteins. See, e.g., GCG version 6.1. Polypeptide sequences can also be compared using FASTA with default or recommended parameters; the FASTA is a program in GCG version 6.1. FASTA (e.g., FASTA2 and FASTA3) provides an alignment of the best overlapping regions between the query and the search sequences and the percentage of sequence identity (Pearson, 2000, supra). When comparing the sequences of the present disclosure with a database containing a large number of sequences from different organisms, another preferred algorithm is the computer program BLAST with default parameters, especially BLASTP or TBLASTN. (See, e.g., Altschul et al., 1990, J. Mol. Biol. 215:403-410 and 1997 Nucleic Acids Res. 25:3389-3402).

[0076] A "variant" of a polypeptide (such as an immunoglobulin, VH, VL, heavy chain, light chain, or CDR containing the amino acid sequences specifically set forth herein) refers to a polypeptide containing an amino acid sequence that is at least about 70%-99.9% identical (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 99.9%) to a reference polypeptide sequence, when compared by the BLAST algorithm, where the parameters of the algorithm are selected to give the maximum match between the corresponding sequences over the entire length of the corresponding reference sequence. In some embodiments, variants of the polypeptide include polypeptides having the amino acid sequence of the reference polypeptide sequence (e.g., as shown in the Sequence Listing below) but having one or more (e.g., 1 to 10, or fewer than 20, or fewer than 10) missense mutations (e.g., conservative substitutions), nonsense mutations, deletions, or insertions.

[0077] The term "therapeutically effective amount" refers to an amount that produces the desired effect upon administration. The exact amount will depend on the therapeutic purpose and can be determined by those skilled in the art using known techniques (see, e.g., Lloyd (1999) The Art, Science and Technology of Pharmaceutical Compounding).

[0078] Anti - Artemin antibodies and antigen - binding fragments thereof

[0079] In one aspect, the present disclosure relates to antibodies that bind artemin and antigen - binding fragments thereof. Artemin can exist in a precursor form (e.g., preproprotein) or a proteolytically processed form, as described, for example, in U.S. Patent No. 6,284,540, which is incorporated herein by reference. In some embodiments, the antibody or its antigen - binding fragment binds to the precursor form of artemin or a fragment thereof. In some embodiments, the antibody or its antigen - binding fragment binds to the proteolytically processed form of artemin or a fragment thereof.

[0080] In some embodiments, the antibody or antigen-binding fragment thereof specifically binds to human artemin (e.g., human preproprotein or proteolytically processed form). In some embodiments, the antibody or antigen-binding fragment thereof specifically binds to human artemin and cross-reacts with artemin from one or more other non-human species, such non-human species including but not limited to mouse, rat, guinea pig, hamster, gerbil, pig, cat, dog, rabbit, goat, sheep, cow, horse, camel, cynomolgus monkey, marmoset, rhesus monkey, or chimpanzee artemin. In some embodiments, the antibody or antigen-binding fragment thereof specifically binds to human artemin and cross-reacts with at least some (e.g., some but not all) non-human forms of artemin (e.g., mouse, rat, guinea pig, hamster, gerbil, pig, cat, dog, rabbit, goat, sheep, cow, horse, camel, cynomolgus monkey, marmoset, rhesus monkey, or chimpanzee artemin). In some embodiments, the antibody or antigen-binding fragment thereof does not cross-react with artemin from non-human species.

[0081] In some embodiments, the anti-artemin antibody or antigen-binding fragment thereof comprises:

[0082] (a) a heavy chain complementarity determining region 1 (HCDR1) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 4, 24, 44, and 64;

[0083] (b) a heavy chain complementarity determining region 2 (HCDR2) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 6, 26, 46, and 66;

[0084] (c) a heavy chain complementarity determining region 3 (HCDR3) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 8, 28, 48, and 68;

[0085] (d) a light chain complementarity determining region 1 (LCDR1) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 12, 32, 52, and 72;

[0086] (e) a light chain complementarity determining region 2 (LCDR2) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 14, 34, 54, and 74; and

[0087] (f) a light chain complementarity determining region 3 (LCDR3) comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 16, 36, 56, and 76.

[0088] In some embodiments, the anti-artemin antibody comprises a heavy chain variable region (HCVR) that comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:2, 22, 42, and 62. In some embodiments, the anti-artemin antibody comprises an HCVR that comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2, 22, 42, and 62.

[0089] In some embodiments, the anti-artemin antibody comprises a light chain variable region (LCVR) that comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:10, 30, 50, and 70. In some embodiments, the anti-artemin antibody comprises an LCVR that comprises an amino acid sequence selected from the group consisting of SEQ ID NO:10, 30, 50, and 70.

[0090] In some embodiments, the anti-artemin antibody or an antigen-binding fragment thereof comprises:

[0091] (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO:4, an HCDR2 comprising the amino acid sequence of SEQ ID NO:6, and an HCDR3 comprising the amino acid sequence of SEQ ID NO:8; or

[0092] (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO:24, an HCDR2 comprising the amino acid sequence of SEQ ID NO:26, and an HCDR3 comprising the amino acid sequence of SEQ ID NO:28; or

[0093] (c) an HCDR1 comprising the amino acid sequence of SEQ ID NO:44, an HCDR2 comprising the amino acid sequence of SEQ ID NO:46, and an HCDR3 comprising the amino acid sequence of SEQ ID NO:48; or

[0094] (d) an HCDR1 comprising the amino acid sequence of SEQ ID NO:64, an HCDR2 comprising the amino acid sequence of SEQ ID NO:66, and an HCDR3 comprising the amino acid sequence of SEQ ID NO:68.

[0095] In some embodiments, the anti-artemin antibody comprises an HCVR that comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:2. In some embodiments, the anti-artemin antibody comprises an HCVR that comprises the amino acid sequence of SEQ ID NO:2. In some embodiments, the anti-artemin antibody comprises an HCVR that comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO:4, an HCDR2 comprising the amino acid sequence of SEQ ID NO:6, and an HCDR3 comprising the amino acid sequence of SEQ ID NO:8, wherein the HCVR has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:2. In some embodiments, the anti-artemin antibody comprises a heavy chain that comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:18. In some embodiments, the anti-artemin antibody comprises a heavy chain that comprises the amino acid sequence of SEQ ID NO:18.

[0096] In some embodiments, the anti-artemin antibody comprises an HCVR that comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:22. In some embodiments, the anti-artemin antibody comprises an HCVR that comprises the amino acid sequence of SEQ ID NO:22. In some embodiments, the anti-artemin antibody comprises an HCVR that comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO:24, an HCDR2 comprising the amino acid sequence of SEQ ID NO:26, and an HCDR3 comprising the amino acid sequence of SEQ ID NO:28, wherein the HCVR has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:22. In some embodiments, the anti-artemin antibody comprises a heavy chain that comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:38. In some embodiments, the anti-artemin antibody comprises a heavy chain that comprises the amino acid sequence of SEQ ID NO:38.

[0097] In some embodiments, the anti-artemin antibody comprises an HCVR that comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:42. In some embodiments, the anti-artemin antibody comprises an HCVR that comprises the amino acid sequence of SEQ ID NO:42. In some embodiments, the anti-artemin antibody comprises an HCVR that comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO:44, an HCDR2 comprising the amino acid sequence of SEQ ID NO:46, and an HCDR3 comprising the amino acid sequence of SEQ ID NO:48, wherein the HCVR has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:42. In some embodiments, the anti-artemin antibody comprises a heavy chain that comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ IDNO:58. In some embodiments, the anti-artemin antibody comprises a heavy chain that comprises the amino acid sequence of SEQ ID NO:58.

[0098] In some embodiments, the anti-artemin antibody comprises an HCVR that comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:62. In some embodiments, the anti-artemin antibody comprises an HCVR that comprises the amino acid sequence of SEQ ID NO:62. In some embodiments, the anti-artemin antibody comprises an HCVR that comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO:64, an HCDR2 comprising the amino acid sequence of SEQ ID NO:66, and an HCDR3 comprising the amino acid sequence of SEQ ID NO:68, wherein the HCVR has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:62. In some embodiments, the anti-artemin antibody comprises a heavy chain that comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:78. In some embodiments, the anti-artemin antibody comprises a heavy chain that comprises the amino acid sequence of SEQ ID NO:78.

[0099] In some embodiments, the anti-artemin antibody or an antigen-binding fragment thereof comprises:

[0100] (a) an LCDR1 comprising the amino acid sequence of SEQ ID NO:12, an LCDR2 comprising the amino acid sequence of SEQ ID NO:14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:16; or

[0101] (b) an LCDR1 comprising the amino acid sequence of SEQ ID NO:32, an LCDR2 comprising the amino acid sequence of SEQ ID NO:34, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:36; or

[0102] (c) an LCDR1 comprising the amino acid sequence of SEQ ID NO:52, an LCDR2 comprising the amino acid sequence of SEQ ID NO:54, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:56; or

[0103] (d) An LCDR1 comprising the amino acid sequence of SEQ ID NO:72, an LCDR2 comprising the amino acid sequence of SEQ ID NO:74, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:76.

[0104] In some embodiments, the anti-artemin antibody comprises an LCVR, which comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:10. In some embodiments, the anti-artemin antibody comprises an LCVR, which comprises the amino acid sequence of SEQ ID NO:10. In some embodiments, the anti-artemin antibody comprises an LCVR, which comprises: an LCDR1 comprising the amino acid sequence of SEQ ID NO:12, an LCDR2 comprising the amino acid sequence of SEQ ID NO:14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:16, wherein the LCVR has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:10. In some embodiments, the anti-artemin antibody comprises a light chain, which comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:20. In some embodiments, the anti-artemin antibody comprises a light chain, which comprises the amino acid sequence of SEQ ID NO:20.

[0105] In some embodiments, the anti-artemin antibody comprises an LCVR, and the LCVR comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity with SEQ ID NO:30. In some embodiments, the anti-artemin antibody comprises an LCVR that comprises the amino acid sequence of SEQ ID NO:30. In some embodiments, the anti-artemin antibody comprises an LCVR that comprises: an LCDR1 comprising the amino acid sequence of SEQ ID NO:32, an LCDR2 comprising the amino acid sequence of SEQ ID NO:34, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:36, wherein the LCVR has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity with SEQ ID NO:30. In some embodiments, the anti-artemin antibody comprises a light chain that comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity with SEQ ID NO:40. In some embodiments, the anti-artemin antibody comprises a light chain that comprises the amino acid sequence of SEQ ID NO:40.

[0106] In some embodiments, the anti-artemin antibody comprises an LCVR, and the LCVR comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:50. In some embodiments, the anti-artemin antibody comprises an LCVR that comprises the amino acid sequence of SEQ ID NO:50. In some embodiments, the anti-artemin antibody comprises an LCVR that comprises: an LCDR1 comprising the amino acid sequence of SEQ ID NO:52, an LCDR2 comprising the amino acid sequence of SEQ ID NO:54, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:56, wherein the LCVR has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:50. In some embodiments, the anti-artemin antibody comprises a light chain that comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:60. In some embodiments, the anti-artemin antibody comprises a light chain that comprises the amino acid sequence of SEQ ID NO:60.

[0107] In some embodiments, the anti-artemin antibody comprises an LCVR, and the LCVR comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:70. In some embodiments, the anti-artemin antibody comprises an LCVR that comprises the amino acid sequence of SEQ ID NO:70. In some embodiments, the anti-artemin antibody comprises an LCVR that comprises: an LCDR1 comprising the amino acid sequence of SEQ ID NO:72, an LCDR2 comprising the amino acid sequence of SEQ ID NO:74, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:76, wherein the LCVR has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:70. In some embodiments, the anti-artemin antibody comprises a light chain that comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:80. In some embodiments, the anti-artemin antibody comprises a light chain that comprises the amino acid sequence of SEQ ID NO:80.

[0108] In some embodiments, the anti-artemin antibody comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO:4, an HCDR2 comprising the amino acid sequence of SEQ ID NO:6, and an HCDR3 comprising the amino acid sequence of SEQ ID NO:8; or an LCDR1 comprising the amino acid sequence of SEQ ID NO:12, an LCDR2 comprising the amino acid sequence of SEQ ID NO:14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:16. In some embodiments, the anti-artemin antibody comprises: an HCVR comprising an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:2, and an LCVR comprising an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:10. In some embodiments, the anti-artemin antibody comprises: an HCVR comprising the amino acid sequence of SEQ ID NO:2, and an LCVR comprising the amino acid sequence of SEQ ID NO:10. In some embodiments, the anti-artemin antibody comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO:18, and a light chain comprising the amino acid sequence of SEQ ID NO:20.

[0109] In some embodiments, the anti-artemin antibody comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO:24, an HCDR2 comprising the amino acid sequence of SEQ ID NO:26, and an HCDR3 comprising the amino acid sequence of SEQ ID NO:28; or an LCDR1 comprising the amino acid sequence of SEQ ID NO:32, an LCDR2 comprising the amino acid sequence of SEQ ID NO:34, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:36. In some embodiments, the anti-artemin antibody comprises: an HCVR comprising an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:22, and an LCVR comprising an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to SEQ ID NO:30. In some embodiments, the anti-artemin antibody comprises: an HCVR comprising the amino acid sequence of SEQ ID NO:22, and an LCVR comprising the amino acid sequence of SEQ ID NO:30. In some embodiments, the anti-artemin antibody comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO:38, and a light chain comprising the amino acid sequence of SEQ ID NO:40.

[0110] In some embodiments, the anti-artemin antibody comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO:44, an HCDR2 comprising the amino acid sequence of SEQ ID NO:46, and an HCDR3 comprising the amino acid sequence of SEQ ID NO:48; or an LCDR1 comprising the amino acid sequence of SEQ ID NO:52, an LCDR2 comprising the amino acid sequence of SEQ ID NO:54, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:56. In some embodiments, the anti-artemin antibody comprises: an HCVR comprising an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity with SEQ ID NO:42, and an LCVR comprising an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity with SEQ ID NO:50. In some embodiments, the anti-artemin antibody comprises: an HCVR comprising the amino acid sequence of SEQ ID NO:42, and an LCVR comprising the amino acid sequence of SEQ ID NO:50. In some embodiments, the anti-artemin antibody comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO:58, and a light chain comprising the amino acid sequence of SEQ ID NO:60.

[0111] In some embodiments, the anti-artemin antibody comprises: an HCDR1 comprising the amino acid sequence of SEQ ID NO:64, an HCDR2 comprising the amino acid sequence of SEQ ID NO:66, and an HCDR3 comprising the amino acid sequence of SEQ ID NO:68; or an LCDR1 comprising the amino acid sequence of SEQ ID NO:72, an LCDR2 comprising the amino acid sequence of SEQ ID NO:74, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:76. In some embodiments, the anti-artemin antibody comprises: an HCVR that comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:62, and an LCVR that comprises an amino acid sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:70. In some embodiments, the anti-artemin antibody comprises: an HCVR that comprises the amino acid sequence of SEQ ID NO:62, and an LCVR that comprises the amino acid sequence of SEQ ID NO:70. In some embodiments, the anti-artemin antibody comprises: a heavy chain that comprises the amino acid sequence of SEQ ID NO:78, and a light chain that comprises the amino acid sequence of SEQ ID NO:80.

[0112] In some embodiments, the antibody or antigen-binding fragment thereof binds artemin in monomeric form. In some embodiments, the antibody or antigen-binding fragment thereof binds artemin in dimeric form. In some embodiments, the antibody or antigen-binding fragment thereof binds the mature form of the artemin protein (e.g., amino acids 108-220 of SEQ ID NO:81).

[0113] In some embodiments, the anti-artemin antibody or antigen-binding fragment of the present disclosure can be linked or co-expressed with another functional molecule (such as another peptide or protein). For example, the antibody or fragment thereof can be functionally linked (e.g., by chemical conjugation, genetic fusion, non-covalent association, or other means) to one or more other molecular entities (such as another antibody or antibody fragment) to produce a bispecific or multispecific antibody having a second binding specificity.

[0114] Sequence variant

[0115] Compared to the corresponding germline sequences from which the individual antibodies are derived, the antibodies or antigen-binding fragments of the present disclosure can contain one or more amino acid substitutions, insertions, and / or deletions in the framework regions and / or CDR regions of the heavy chain variable domain and the light chain variable domain. Such mutations can be readily determined by comparing the amino acid sequences disclosed herein with germline sequences obtainable from, for example, public antibody sequence databases. The antibodies of the present disclosure can contain antigen-binding fragments derived from any of the exemplary amino acid sequences disclosed herein, wherein one or more amino acids within one or more frameworks and / or CDRs are mutated to the corresponding residues of the germline sequence from which the antibody was derived, or to the corresponding residues of another human germline sequence, or to conservative amino acid substitutions of the corresponding germline residues (such sequence variations are collectively referred to herein as "germline mutations"). One of ordinary skill in the art can readily generate a variety of antibodies and antigen-binding fragments starting from the heavy and light chain variable region sequences disclosed herein, which contain one or more individual germline mutations or combinations thereof. In certain embodiments, all framework and / or CDR residues within the V H and / or V L domains are mutated back to the residues found in the original germline sequence from which the antibody was originally derived. In other embodiments, only certain residues are mutated back to the original germline sequence, e.g., only the mutated residues present within the first 8 amino acids of FR1 or the last 8 amino acids of FR4, or only the mutated residues present within CDR1, CDR2, or CDR3. In other embodiments, one or more of the framework and / or one or more CDR residues are mutated to one or more corresponding residues of a different germline sequence (i.e., a germline sequence different from the germline sequence from which the antibody was originally derived). Additionally, the antibody or antigen-binding fragment can contain any combination of two or more germline mutations within the framework and / or CDR regions, e.g., wherein certain individual residues are mutated to the corresponding residues of a specific germline sequence, while certain other residues different from the original germline sequence are either maintained or mutated to the corresponding residues in a different germline sequence. After obtaining an antibody or antigen-binding fragment containing one or more germline mutations, one or more desired properties of the antibody and antigen-binding fragment can be readily tested, such as improved binding specificity, increased binding affinity, improved or enhanced antagonistic or agonistic biological properties, reduced immunogenicity, etc. Antibodies and antigen-binding fragments obtained in this general manner are encompassed within the present disclosure.

[0116] The present disclosure also includes antibodies or antigen-binding fragments that comprise variants of any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein having one or more conservative substitutions. For example, the present disclosure includes antibodies or antigen-binding fragments that comprise HCVR, LCVR, and / or CDR amino acid sequences having, for example, 10 or fewer, 9 or fewer, 8 or fewer, 7 or fewer, 6 or fewer, 5 or fewer, 4 or fewer, 3 or fewer, 2 or fewer, or 1 conservative amino acid substitution relative to any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein. A "conservative amino acid substitution" is a substitution in which one amino acid residue is replaced with another amino acid residue having a side chain (R-group) with similar chemical properties (e.g., charge or hydrophobicity). In general, conservative amino acid substitutions do not substantially alter the functional properties of the protein. Examples of groups of amino acids having side chains with similar chemical properties include: (1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; (2) aliphatic-hydroxyl side chains: serine and threonine; (3) amide-containing side chains: asparagine and glutamine; (4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; (5) basic side chains: lysine, arginine, and histidine; (6) acidic side chains: aspartic acid and glutamic acid; and (7) sulfur-containing side chains, namely cysteine and methionine. Preferred groups of conservative amino acid substitutions are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamic acid-aspartic acid, and asparagine-glutamine. Alternatively, conservative substitutions are any changes having a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al. (1992) Science 256:1443-1445. A "moderately conservative" substitution is any change having a non-negative value in the PAM250 log-likelihood matrix.

[0117] The present disclosure also includes antibodies or antigen-binding fragments thereof that comprise HCVR, LCVR, and / or CDR amino acid sequences that are substantially identical to any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein. In some embodiments, the antigen-binding molecule comprises an HCVR, LCVR, and / or CDR amino acid sequence having at least 85% sequence identity, such as at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity, with the sequences disclosed in Table 1, Table 2, or Table 9. In some embodiments, the antigen-binding molecule comprises an HCVR, LCVR, and / or CDR amino acid sequence having at least 85% sequence identity, such as at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity, with the sequences disclosed in Table 1, Table 2, or Table 9, wherein the differences in amino acid residues relative to the sequences disclosed in Table 1, Table 2, or Table 9 are conservative substitutions or moderately conservative substitutions.

[0118] In some embodiments, the antibody or antigen-binding fragment thereof is chimeric, humanized, or fully human. In some embodiments, the antibody or antigen-binding fragment thereof is humanized. In some embodiments, the antibody or antigen-binding fragment thereof is fully human.

[0119] Antibody comprising an Fc variant

[0120] In some embodiments, anti-artemin antibodies or antigen-binding fragments thereof are provided that comprise an Fc domain, the Fc domain comprising one or more mutations that enhance or attenuate antibody binding to the FcRn receptor at an acidic pH compared to neutral pH. For example, the present disclosure includes antibodies or antigen-binding fragments that comprise one or more mutations in the CH2 and / or CH3 regions of the Fc domain, wherein the mutations increase the affinity of the Fc domain for FcRn in an acidic environment (e.g., in an endosome at a pH range of about 5.5 to about 6.0). When administered to an animal, such mutations can result in an increased serum half-life of the antibody. Non-limiting examples of such Fc modifications include, for example, modifications at position 250 (e.g., E or Q); 250 and 428 (e.g., L or F); 252 (e.g., L / Y / F / W or T), 254 (e.g., S or T), and 256 (e.g., S / R / Q / E / D or T); or modifications at position 428 and / or 433 (e.g., H / L / R / S / P / Q or K) and / or 434 (e.g., H / F or Y); or modifications at position 250 and / or 428; or modifications at position 307 or 308 (e.g., 308F, V308F) and 434. In one embodiment, the modification comprises 428L (e.g., M428L) and 434S (e.g., N434S) modifications; 428L, 259I (e.g., V259I), and 308F (e.g., V308F) modifications; 433K (e.g., H433K) and 434 (e.g., 434Y) modifications; 252, 254, and 256 (e.g., 252Y, 254T, and 256E) modifications; 250Q and 428L modifications (e.g., T250Q and M428L); and 307 and / or 308 modifications (e.g., 308F or 308P).

[0121] As non-limiting examples, the present disclosure includes anti-artemin antibodies that comprise an Fc domain, the Fc domain comprising one or more pairs or groups of mutations selected from the group consisting of: 250Q and 248L (e.g., T250Q and M248L); 252Y, 254T, and 256E (e.g., M252Y, S254T, and T256E); 428L and 434S (e.g., M428L and N434S); and 433K and 434F (e.g., H433K and N434F). All possible combinations of the foregoing Fc domain mutations and other mutations within the antibody variable domains disclosed herein are encompassed by the present disclosure.

[0122] Polynucleotides, vectors, and host cells

[0123] In another aspect, the present disclosure provides nucleic acid molecules comprising one or more polynucleotide sequences encoding the antibodies or antigen-binding fragments disclosed herein, as well as vectors (e.g., expression vectors) encoding such polynucleotide sequences and host cells into which such vectors have been introduced.

[0124] In some embodiments, the nucleic acid molecule comprises one or more polynucleotide sequences encoding the antibodies or antigen-binding fragments disclosed in Table 1 or Table 9. In some embodiments, the nucleic acid molecule comprises one or more polynucleotide sequences shown in Table 2 or Table 9.

[0125] In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an HCVR comprising HCDR1, HCDR2, and HCDR3 comprising SEQ ID NO: 4, 6, and 8, respectively. In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an HCVR comprising the sequence of SEQ ID NO: 2. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequences of SEQ ID NO: 3, 5, and 7. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO: 1 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO: 1. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO: 17 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO: 17.

[0126] In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an HCVR that respectively comprises HCDR1, HCDR2, and HCDR3 of SEQ ID NO: 24, 26, and 28. In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an HCVR that comprises the sequence of SEQ ID NO: 22. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequences of SEQ ID NO: 23, 25, and 27. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO: 21 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO: 21. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO: 37 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO: 37.

[0127] In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an HCVR that respectively comprises HCDR1, HCDR2, and HCDR3 of SEQ ID NO: 44, 46, and 48. In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an HCVR that comprises the sequence of SEQ ID NO: 42. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequences of SEQ ID NO: 43, 45, and 47. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO: 41 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO: 41. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO: 57 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO: 57.

[0128] In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an HCVR that encodes HCDR1, HCDR2, and HCDR3 comprising SEQ ID NO: 64, 66, and 68, respectively. In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an HCVR comprising the sequence of SEQ ID NO: 62. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequences of SEQ ID NO: 63, 65, and 67. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO: 61 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO: 61. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO: 77 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO: 77.

[0129] In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an LCVR that encodes LCDR1, LCDR2, and LCDR3 comprising SEQ ID NO: 12, 14, and 16, respectively. In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an LCVR comprising the sequence of SEQ ID NO: 10. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequences of SEQ ID NO: 11, 13, and 15. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO: 9 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO: 9. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO: 19 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO: 19.

[0130] In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an LCVR that comprises LCDR1, LCDR2, and LCDR3 that comprise SEQ ID NO: 32, 34, and 36, respectively. In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an LCVR that comprises the sequence of SEQ ID NO: 30. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequences of SEQ ID NO: 31, 33, and 35. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO: 29 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO: 29. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO: 39 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO: 39.

[0131] In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an LCVR that comprises LCDR1, LCDR2, and LCDR3 that comprise SEQ ID NO: 52, 54, and 56, respectively. In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an LCVR that comprises the sequence of SEQ ID NO: 50. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequences of SEQ ID NO: 51, 53, and 55. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO: 49 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO: 49. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO: 59 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO: 59.

[0132] In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an LCVR comprising LCDR1, LCDR2, and LCDR3 comprising SEQ ID NO:72, 74, and 76, respectively. In some embodiments, the nucleic acid molecule comprises a polynucleotide sequence encoding an LCVR comprising the sequence of SEQ ID NO:70. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequences of SEQ ID NO:71, 73, and 75. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO:69 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO:69. In some embodiments, the nucleic acid molecule comprises the polynucleotide sequence of SEQ ID NO:79 or has at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) with SEQ ID NO:79.

[0133] In some embodiments, a composition is provided that comprises one or more of the nucleic acid molecules disclosed herein (e.g., a first nucleic acid molecule comprising a polynucleotide sequence encoding an HCVR of an anti-artemin antibody, and a second nucleic acid molecule comprising a polynucleotide sequence encoding an LCVR of an anti-artemin antibody).

[0134] The present disclosure also provides recombinant expression vectors carrying one or more of the nucleic acid molecules disclosed herein, and host cells into which such vectors have been introduced. In some embodiments, two or more expression vectors are provided (e.g., a first expression vector comprising a first nucleic acid molecule that comprises a polynucleotide sequence encoding an HCVR of an anti-artemin antibody, and a second expression vector comprising a second nucleic acid molecule that comprises a polynucleotide sequence encoding an LCVR of an anti-artemin antibody). In some embodiments, the expression vector comprises two or more nucleic acid molecules (e.g., the expression vector comprises (i) a first nucleic acid molecule that comprises a polynucleotide sequence encoding an HCVR of an anti-artemin antibody, and (ii) a second nucleic acid molecule that comprises a polynucleotide sequence encoding an LCVR of an anti-artemin antibody). Also provided herein are methods of producing an anti-artemin antibody or antigen-binding fragment using the nucleic acid sequences and / or vectors described herein. In some embodiments, the method of producing an antibody or its antigen-binding fragment comprises culturing a host cell comprising one or more of the nucleic acid sequences and / or expression vectors described herein under conditions that permit the production of the antibody or antigen-binding fragment, and recovering the antibody or antigen-binding fragment so produced.

[0135] In some embodiments, the host cell comprising one or more of the nucleic acid sequences and / or vectors described herein is a prokaryotic cell (e.g., Escherichia coli (E. coli)). In some embodiments, the host cell is a eukaryotic cell, such as a non-human mammalian cell (e.g., Chinese hamster ovary (CHO) cell, NS0 cell, BHK cell, or HEK293 cell). Also provided herein is a method for producing an anti-artemin antibody or antigen-binding fragment by culturing the host cell under conditions permitting the production of the antibody or antigen-binding fragment, and recovering the antibody or antigen-binding fragment so produced.

[0136] Characterization of Anti-Artemin Antibodies

[0137] The present disclosure includes antibodies and antigen-binding fragments thereof that bind human artemin with high affinity. In some embodiments, the present disclosure includes antibodies and antigen-binding fragments thereof that bind human ARTN (e.g., at 25 °C) with a K D as measured by surface plasmon resonance or biolayer interferometry, e.g., using the assay format defined in Example 2 herein. In some embodiments, the human ARTN is the mature form of ARTN, e.g., amino acids 108 - 220 of the human ARTN protein sequence shown in SEQ ID NO:81. In certain embodiments, the antibodies or antigen-binding fragments of the present disclosure bind human ARTN with a K of less than about 5 nM, less than about 1 nM, less than about 500 pM, less than about 400 pM, less than about 300 pM, less than about 200 pM, less than about 100 pM, less than about 90 pM, less than about 80 pM, less than about 70 pM, less than about 60 pM, less than about 50 pM, less than about 40 pM, less than about 30 pM, less than about 20 pM, less than about 10 pM, less than about 5 pM, less than about 4 pM, less than about 2 pM, less than about 1 pM, less than about 0.5 pM, less than about 0.2 pM, less than about 0.1 pM, or less than about 0.05 pM. D bind human ARTN, as measured by surface plasmon resonance, e.g., using the assay format defined in Example 2 herein or a substantially similar assay.

[0138] In some embodiments, the present disclosure includes antibodies and antigen-binding fragments thereof that bind to human ARTN with a dissociation half-life (t1 / 2) greater than about 30 minutes, as measured by surface plasmon resonance or biolayer interferometry at 25°C, for example using the assay format defined in Example 2 herein or substantially similar assays. In certain embodiments, the antibodies or antigen-binding fragments of the present disclosure bind to human ARTN with a dissociation half-life (t1 / 2) greater than about 30 minutes, greater than about 40 minutes, greater than about 50 minutes, greater than about 60 minutes, greater than about 70 minutes, greater than about 80 minutes, greater than about 90 minutes, greater than about 100 minutes, greater than about 150 minutes, greater than about 200 minutes, greater than about 250 minutes, or greater than about 300 minutes, as measured by surface plasmon resonance at 25°C, for example using the assay format defined in Example 2 herein or substantially similar assays.

[0139] In some embodiments, the present disclosure includes antibodies and antigen-binding fragments thereof that block the binding of ARTN protein to GFRα3. In some embodiments, the present disclosure includes antibodies and antigen-binding fragments thereof that block the binding of monomeric ARTN (e.g., human, cynomolgus monkey, and / or mouse ARTN) protein to human GFRα3, as measured by a blocking ELISA assay, for example using the assay format defined in Example 2 herein or substantially similar assays. In certain embodiments, the antibodies or antigen-binding fragments of the present disclosure block at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90% of the binding of human ARTN to human GFRα3, as measured by a blocking ELISA assay, for example using the assay format defined in Example 3 herein or substantially similar assays.

[0140] In some embodiments, the present disclosure includes antibodies and antigen-binding fragments thereof that inhibit artemin-mediated cell signaling. In some embodiments, the present disclosure includes antibodies and antigen-binding fragments thereof that inhibit artemin-mediated cell signaling, as measured using a biological assay in a cell line expressing human GFRα3 and human RET, for example, as described in Example 4 or substantially similar assays.

[0141] Epitope mapping and related techniques

[0142] In some embodiments, the epitope bound by an antibody of the present disclosure can consist of a single contiguous sequence of amino acids of 3 or more (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more) ARTN proteins. Alternatively, the epitope can consist of non-contiguous amino acids (or amino acid sequences) of multiple ARTNs. In some embodiments, the antibodies of the present disclosure can interact with the amino acids contained in an ARTN monomer or can interact with amino acids on two different chains of an ARTN dimer. As used herein, the term "epitope" refers to an antigenic determinant that interacts with a specific antigen-binding site (termed a paratope) in the variable region of an antibody molecule. A single antigen can have more than one epitope. Thus, different antibodies can bind to different regions on an antigen and can have different biological effects. Epitopes can be conformational or linear. Conformational epitopes are generated by the spatial juxtaposition of amino acids from different segments of a linear polypeptide chain. Linear epitopes are generated by neighboring amino acid residues in a polypeptide chain. In some cases, an epitope can include a sugar, phosphoryl group or sulfonyl group moiety on the antigen.

[0143] A variety of techniques known to those of ordinary skill in the art can be used to determine whether an antibody "interacts with one or more amino acids" within a polypeptide or protein. Exemplary techniques that can be used to determine the epitope or binding domain of a particular antibody include, for example, conventional cross-blocking assays (such as those described in Antibodies, Harlow and Lane (Cold Spring Harbor Press, Cold Spring Harbor, NY)), site-directed mutagenesis (e.g., alanine-scanning mutagenesis, arginine-scanning mutagenesis, etc.), peptide blotting (Reineke, 2004, Methods Mol Biol 248:443-463), protease protection, and peptide cleavage analysis. In addition, methods such as epitope excision, epitope extraction, and chemical modification of the antigen can be used (Tomer, 2000, Protein Science 9:487-496). Another method that can be used to identify the amino acids within a polypeptide that interact with an antibody is hydrogen / deuterium exchange detected by mass spectrometry. Generally, the hydrogen / deuterium exchange method involves deuterium-labeling the protein of interest and then allowing the antibody to bind to the deuterium-labeled protein. The protein / antibody complex is then transferred to water to allow hydrogen-deuterium exchange to occur at all residues except those protected by the antibody (which remain deuterium-labeled). After antibody dissociation, the target protein is subjected to protease cleavage and mass spectrometry, which reveals the deuterium-labeled residues that correspond to the specific amino acids that interact with the antibody. See, for example, Ehring (1999) Analytical Biochemistry 267(2):252-259; Engen and Smith (2001) Anal. Chem. 73:256A-265A. X-ray crystallographic analysis can also be used to identify the amino acids within a polypeptide with which an antibody interacts.

[0144] In some embodiments, the present disclosure provides an anti-artemin antibody that binds to the same epitope as any of the specific exemplary antibodies described herein (e.g., an antibody comprising any of the amino acid sequences shown in Table 1 or Table 9 below).

[0145] In some embodiments, the present disclosure provides an anti-artemin antibody that competes with any of the specific exemplary anti-artemin antibodies described herein for binding to ARTN.

[0146] Using conventional methods known in the art, one of ordinary skill in the art can determine whether a particular antibody binds to the same epitope as a reference antibody of the present disclosure or whether it competes with the reference antibody of the present disclosure for binding. For example, to determine whether a test antibody binds to the same epitope on ARTN as a reference antibody of the present disclosure, first allow the reference antibody to bind to the ARTN protein. Then, the ability of the test antibody to bind to the ARTN protein is evaluated. If the test antibody is able to bind to ARTN after saturation binding with the reference antibody, it can be concluded that the test antibody binds to a different epitope than the reference antibody. In another aspect, if the test antibody is unable to bind to ARTN after saturation binding with the reference antibody, the test antibody may bind to the same epitope on ARTN as the epitope to which the reference antibody of the present disclosure binds. Then additional routine experiments (e.g., peptide mutagenesis and binding assays) can be performed to confirm whether the observed lack of binding of the test antibody is actually due to binding to the same epitope as the reference antibody or whether it is due to steric hindrance (or another phenomenon) resulting in the observed lack of binding. Such experiments can be performed using ELISA, RIA, Biacore, flow cytometry, or any other quantitative or qualitative antibody binding assay available in the art. According to certain embodiments of the present disclosure, if, for example, a 1-fold, 2-fold, 5-fold, 10-fold, 20-fold, or 100-fold excess of one antibody inhibits the binding of another antibody by at least 50%, but preferably 75%, 90%, or even 99%, as measured in a competitive binding assay, the two antibodies bind to the same (or overlapping) epitope (see, e.g., Junghans et al., Cancer Res. 1990:50:1495-1502). Alternatively, if substantially all of the amino acid mutations in the antigen that reduce or eliminate the binding of one antibody also reduce or eliminate the binding of the other antibody, the two antibodies are considered to bind to the same epitope. If only a subset of the amino acid mutations that reduce or eliminate the binding of one antibody also reduce or eliminate the binding of the other antibody, the two antibodies are considered to have "overlapping epitopes".

[0147] To determine whether a test antibody or its antigen-binding fragment competes with a reference antibody for binding, the above-described binding method is performed in two directions: in the first direction, allow the reference antibody to bind to the ARTN protein under saturation conditions, and then evaluate the binding of the test antibody to the ARTN protein. In the second direction, allow the test antibody to bind to the ARTN protein under saturation conditions, and then evaluate the binding of the reference antibody to the ARTN protein. If in both directions only the first (saturated) antibody is able to bind to the ARTN molecule, it can be concluded that the test antibody and the reference antibody compete for binding to ARTN. As would be understood by one of ordinary skill in the art, an antibody that competes with a reference antibody for binding may not necessarily bind to the same epitope as the reference antibody, but may sterically block the binding of the reference antibody by binding to an overlapping or adjacent epitope.

[0148] Preparation of Antibodies

[0149] The antibodies disclosed herein can be prepared by any antibody production techniques known in the art. In certain embodiments, one or more individual components of the antibody (e.g., heavy and light chains) are derived from chimeric, humanized, or fully human antibodies. Methods for preparing such antibodies are well known in the art. For example, VELOCIMMUNE TM technology can be used to prepare one or more heavy chains and / or light chains of the antibodies disclosed herein. Using VELOCIMMUNE TM technology (or any other technology for generating human antibodies), high-affinity chimeric antibodies specific for a particular antigen (e.g., ARTN) with human variable regions and murine constant regions are first isolated. Antibodies are characterized and selected for desired characteristics, including affinity, selectivity, epitope, etc. The murine constant regions are replaced with the desired human constant regions to generate fully human heavy chains and / or light chains that can be incorporated into the antibody.

[0150] In some embodiments, antibodies can be generated by administering an immunogen (e.g., a peptide as shown in SEQ ID NO:82, optionally conjugated to a carrier) to a mouse, optionally in combination with an adjuvant. After immunization, spleen cells are harvested and fused with mouse myeloma cells to maintain their viability and form a hybridoma cell line. The hybridoma cell line is screened and selected to identify the cell line that produces artemin-specific antibodies. Alternatively, DNA encoding antigen-specific chimeric antibodies or variable domains of light and heavy chains can be directly isolated from antigen-positive B cells without fusion with myeloma cells, as described in U.S. 2007 / 0280945 or WO2016077666, both of which are incorporated herein by reference in their entireties.

[0151] In some embodiments, genetically engineered animals can be used to produce human antibodies. Non-limiting exemplary genetically modified mice and methods for generating antibodies from genetically modified mice are described in US 8,697,940, US10,130,081, US10,561,124, and US10,640,800, the entire contents of which are incorporated herein by reference. As used herein, "fully human" refers to an antigen-binding molecule, such as an antibody or an antigen-binding fragment thereof or an immunoglobulin domain, that contains an amino acid sequence encoded by DNA from human sequences over the full length of each polypeptide of the antigen-binding molecule, antibody, antigen-binding fragment, or its immunoglobulin domain. In some cases, the fully human sequence is derived from an endogenous protein of a human. In other cases, the fully human protein or protein sequence contains chimeric sequences, where each component sequence is from a human sequence. Although not bound by any one theory, chimeric proteins or chimeric sequences are generally designed, for example, to minimize the generation of immunogenic epitopes at the junctions of the component sequences compared to any wild-type human immunoglobulin region or domain.

[0152] Bioequivalent

[0153] The present disclosure includes antibodies having an amino acid sequence different from the antibody but retaining the ability to bind ARTN. Such variant molecules contain one or more additions, deletions, or substitutions of amino acids compared to the parental sequence, but exhibit a biological activity that is substantially equivalent to the biological activity of the antibody. Similarly, nucleic acid sequences encoding the antibodies of the present disclosure include sequences that contain one or more nucleotide additions, deletions, or substitutions compared to the disclosed sequences, but encode antibodies that are substantially bioequivalent to the antibodies disclosed herein.

[0154] The present disclosure includes antibodies that are bioequivalent to any of the exemplary antibodies set forth herein. For example, two antibodies are considered bioequivalent if they are pharmaceutical equivalents or drug substitutes, i.e., when administered at the same molar dose (single dose or multiple doses) under similar experimental conditions, their rates and extents of absorption do not show significant differences. If some antibodies are the same in extent of absorption but different in rate of absorption, they may be considered equivalents or drug substitutes and still be considered bioequivalent because such differences in rate of absorption are intentional and reflected in the specification and are not required for achieving an effective in vivo drug concentration (e.g., in long-term use) and are considered medically insignificant for the particular drug product being studied.

[0155] In one embodiment, two antibodies are bioequivalents if there are no clinically significant differences in terms of safety, purity, and potency.

[0156] In one embodiment, two antibodies are bioequivalent if a patient can make such a switch one or more times without an increased risk of an expected adverse reaction, including a clinically significant change in immunogenicity or decreased efficacy, compared to a continuous therapy where no switch is made between a first antibody (e.g., a reference product) and a second antibody (e.g., a biological product).

[0157] In one embodiment, two antibodies are bioequivalent if both antibodies act through one or more common mechanisms of action for one or more conditions of use to the extent that such mechanisms are known.

[0158] Bioequivalence can be demonstrated by in vivo and in vitro methods. Non-limiting examples of bioequivalence metrics include, for example, (a) in vivo testing in humans or other mammals where the concentration of the antibody or its metabolites in blood, plasma, serum, or other biological fluids is measured over time; (b) in vitro testing that is correlated with in vivo bioavailability data in humans and can reasonably predict in vivo bioavailability data; (c) in vivo testing in humans or other mammals where the appropriate acute pharmacological effect of the antibody (or its target) is measured over time; and (d) well-controlled clinical trials that establish the safety, efficacy, or bioavailability or bioequivalence of the antibody.

[0159] Bioequivalent variants of the exemplary antibodies described herein can be constructed, for example, by making various substitutions of residues or sequences or by deleting terminal or internal residues or sequences that are not required for biological activity. For example, cysteine residues that are not essential for biological activity can be deleted or replaced with other amino acids to prevent the formation of unnecessary or incorrect intramolecular disulfide bonds upon refolding. In other embodiments, bioequivalent antibodies can include the exemplary antibodies described herein that contain amino acid changes that alter the glycosylation profile of the antibody, such as mutations that eliminate or remove glycosylation.

[0160] Pharmaceutical Compositions

[0161] In another aspect, the present disclosure provides pharmaceutical compositions comprising the anti-artemin antibodies and antigen-binding fragments disclosed herein. The pharmaceutical compositions are formulated with one or more pharmaceutically acceptable excipients, carriers, and / or diluents. A variety of pharmaceutically acceptable carriers and diluents are well known in the art (see, e.g., Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA). In some embodiments, the carrier is suitable for intravenous, intramuscular, oral, intraperitoneal, intrathecal, percutaneous, topical, or subcutaneous administration.

[0162] In some embodiments, the pharmaceutical composition comprises an injectable preparation, such as a dosage form for intravenous, subcutaneous, intradermal, and intramuscular injection, infusion, etc. These injectable preparations can be prepared by known methods. For example, an injectable preparation can be prepared as follows: For example, the above-mentioned antibody or its salt is dissolved, suspended, or emulsified in a sterile aqueous medium or an oily medium conventionally used for injections. As the aqueous medium for injection, there are, for example, physiological saline, isotonic solutions containing glucose and other adjuvants, etc., which can be used in combination with a suitable solubilizer, such as an alcohol (e.g., ethanol), a polyol (e.g., propylene glycol, polyethylene glycol), a nonionic surfactant [e.g., polysorbate 80, HCO-50 (hydrogenated castor oil polyoxyethylene (50 mol) adduct)], etc. As the oily medium, for example, sesame oil, soybean oil, etc. are used, which can be used in combination with a solubilizer, such as benzyl benzoate, benzyl alcohol, etc. The injectable preparation thus prepared can be filled into a suitable ampoule.

[0163] The dosage of the antibody administered to a patient according to the present disclosure can vary depending on the patient's age and body size, symptoms, condition, route of administration, etc. The dosage is usually calculated based on body weight or body surface area. Depending on the severity of the disease condition, the frequency and duration of treatment can be adjusted. The effective dosage and regimen for administering the pharmaceutical composition disclosed herein can be determined empirically; for example, the patient's progress can be monitored by regular evaluations, and the dosage can be adjusted accordingly. In addition, interspecies scaling of the dosage can be performed using methods well known in the art (e.g., Mordenti et al., 1991, Pharmaceut. Res. 8:1351).

[0164] Various delivery systems are known and can be used to administer the pharmaceutical composition, such as encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, receptor-mediated endosomes (see, for example, Wu et al., 1987, J. Biol. Chem. 262:4429-4432). The methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The composition can be administered by any convenient route, such as by infusion or bolus injection, by absorption through the epithelial or mucosal skin layers (e.g., oral mucosa, rectal and intestinal mucosa, etc.), and can be administered together with other bioactive agents. In some embodiments, the pharmaceutical composition as disclosed herein is administered intravenously. In some embodiments, the pharmaceutical composition as disclosed herein is administered subcutaneously.

[0165] In some embodiments, the antibodies or antibody-containing pharmaceutical compositions disclosed herein are contained within a container. Accordingly, in another aspect, provided is a container comprising the antibodies or pharmaceutical compositions disclosed herein. For example, in some embodiments, the antibody or pharmaceutical composition is contained in a container selected from the group consisting of glass vials, syringes, pen delivery devices, and autoinjectors.

[0166] In some embodiments, the antibodies or pharmaceutical compositions of the present disclosure are delivered subcutaneously or intravenously using a standard needle and syringe. In some embodiments, the syringe is a pre-filled syringe. In some embodiments, a pen delivery device or autoinjector is used to deliver the antibodies or pharmaceutical compositions of the present disclosure (e.g., for subcutaneous delivery). The pen delivery device can be reusable or disposable. Reusable pen delivery devices typically utilize a replaceable cartridge containing the pharmaceutical composition. Once all of the pharmaceutical composition in the cartridge has been administered and the cartridge is empty, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. The pen delivery device can then be reused. In a disposable pen delivery device, there is no replaceable cartridge. Rather, the disposable pen delivery device is pre-filled with the pharmaceutical composition held in a reservoir within the device. Once the pharmaceutical composition in the reservoir is emptied, the entire device is discarded.

[0167] Examples of suitable pen and autoinjector delivery devices include, but are not limited to, AUTOPEN TM (Owen Mumford, Inc., Woodstock, UK), DISETRONIC TM Pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75 / 25 TM Pen, HUMALOG TM Pen, HUMALIN 70 / 30 TM Pen (Eli Lilly and Co., Indianapolis, IN), NOVOPEN TM I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR TM (Novo Nordisk, Copenhagen, Denmark), BD TM Pen (Becton Dickinson, Franklin Lakes, NJ), OPTIPEN TM , OPTIPEN PRO TM , OPTIPEN STARLETTM and OPTICLIK TM (sanofi - aventis, Frankfurt, Germany). Examples of single - use pen - type delivery devices that can be used for subcutaneous delivery of the pharmaceutical compositions of the present invention include, but are not limited to, SOLOSTAR TM Pen (sanofi - aventis), FLEXPEN TM (Novo Nordisk) and KWIKPEN TM (Eli Lilly), SURECLICK TM Autoinjector (Amgen, Thousand Oaks, CA), PENLET TM (Haselmeier, Stuttgart, Germany), EPIPEN (Dey, L.P.) and HUMIRA TM Pen (Abbott Labs, Abbott Park IL).

[0168] In some embodiments, an antibody or pharmaceutical composition is delivered using a controlled - release system. In one embodiment, a pump can be used (see Langer, supra; Sefton, 1987, CRC Crit. Ref. Biomed. Eng. 14:201). In another embodiment, a polymeric material can be used; see Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Pres., Boca Raton, Florida. In yet another embodiment, the controlled - release system can be placed near the target of the composition, so that only a small fraction of the systemic dose is required (see, for example, Goodson, 1984, in Medical Applications of Controlled Release, supra, Volume 2, pp. 115 - 138). Other controlled - release systems are discussed in the review by Langer, 1990, Science, 249:1527 - 1533.

[0169] In some embodiments, the pharmaceutical compositions for use as described herein are formulated into dosage forms in unit doses suitable for the dose of the active ingredient. Such unit - dose dosage forms include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc. In some embodiments, the amount of the antigen - binding molecule contained in the dosage form is from about 5 mg to about 500 mg, such as from about 5 mg to about 100 mg or from about 10 mg to about 250 mg.

[0170] Therapeutic Use

[0171] In another aspect, the present disclosure provides methods of using the anti-artemin antibodies and antigen-binding fragments disclosed herein. In some embodiments, the anti-artemin antibodies and antigen-binding fragments disclosed herein can be used to treat, prevent, and / or ameliorate any disease, disorder, or condition associated with artemin expression (e.g., overexpression) or activity. In some embodiments, the disease, disorder, or condition is pain, such as acute pain, chronic pain, neuropathic pain, inflammatory pain, neuralgia (e.g., trigeminal neuralgia, postherpetic neuralgia, or general neuralgia), visceral pain, osteoarthritis pain, gout, radicular pain, sciatica, back pain, head or neck pain, breakthrough pain, postoperative pain, chemotherapy-induced neuropathic pain, radiotherapy-induced neuropathic pain, radiation-related pain, or cancer pain (e.g., pain associated with bone cancer or pancreatic cancer).

[0172] In some embodiments, the anti-artemin antibodies and antigen-binding fragments disclosed herein can be used to treat, prevent, and / or ameliorate headache or pain associated with headache (e.g., for migraine, cluster headache, chronic headache, or tension headache).

[0173] In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein can also be used to treat any of the following conditions: non-malignant acute, chronic, or fracture-related bone pain; pain associated with arthritis (e.g., osteoarthritis or rheumatoid arthritis); spinal stenosis; neuropathic low back pain; myofascial pain syndrome; fibromyalgia; temporomandibular joint pain; visceral pain; chronic headache pain; tension headache, including cluster headache; migraine; diabetic neuropathy; HIV-related neuropathy; Charcot-Marie Tooth neuropathy; hereditary sensory neuropathy; peripheral nerve injury; painful neuroma; ectopic proximal and distal discharges; radiculopathy; chemotherapy-induced neuropathic pain; radiotherapy-induced neuropathic pain; post-mastectomy pain; central pain; spinal cord injury pain; post-stroke pain; thalamic pain; complex regional pain syndrome (CRPS); phantom pain; intractable pain; musculoskeletal pain; joint pain; acute gout pain; pruritus; mechanical low back pain; neck pain; tendinitis; injury / sports pain; abdominal pain; chest pain, including cardiogenic pain; pelvic pain; renal colic; acute obstetric pain, including labor pain; cesarean section pain; burn and trauma pain; endometriosis; post-herpetic neuralgia; breakthrough pain; orofacial pain, including sinus pain or toothache; pain in multiple sclerosis; leprosy pain; Guillain-Barre pain; and Burning Mouth Syndrome.

[0174] Dosage and Administration Regimen

[0175] In some embodiments, the amount of the anti-artemin antibody or antigen-binding fragment administered to a subject according to the methods disclosed herein is a therapeutically effective amount. As used herein, the phrase "therapeutically effective amount" refers to the amount that produces the desired effect upon administration.

[0176] In some embodiments, the antibody is administered to a subject at a body weight-based dose. A "body weight-based dose" (e.g., a dose in mg / kg) is a dose of the antibody that will vary depending on the body weight of the subject.

[0177] In other embodiments, the antibody is administered at a fixed dose. A "fixed dose" (e.g., a dose in mg) refers to a single dose of the antibody for all subjects, regardless of any specific subject-related factors such as body weight. In a particular embodiment, the fixed dose of the antibody is based on a predetermined body weight or age.

[0178] Typically, a suitable dose of the antibody can range from about 0.001 mg / kg to about 200.0 mg / kg of recipient body weight, usually in the range of about 1 mg / kg to 50 mg / kg of body weight. For example, the antibody can be administered at a dose of about 0.1 mg / kg, about 0.2 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 1.5 mg / kg, about 2 mg / kg, about 3 mg / kg, about 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 40 mg / kg, or about 50 mg / kg per single dose. Intermediate values and ranges of these values are also part of the present disclosure.

[0179] In some embodiments, the antibody is administered at a fixed dose between about 5 mg and about 2500 mg. In some embodiments, the antibody is administered at a fixed dose of about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg, about 500 mg, about 525 mg, about 550 mg, about 575 mg, about 600 mg, about 625 mg, about 650 mg, about 675 mg, about 700 mg, about 725 mg, about 750 mg, about 775 mg, about 800 mg, about 825 mg, about 850 mg, about 875 mg, about 900 mg, about 925 mg, about 950 mg, about 975 mg, about 1000 mg, about 1500 mg, about 2000 mg, or about 2500 mg. Intermediate values and ranges of these values are also part of the present disclosure.

[0180] In some embodiments, the antibody is administered to the subject at a dosing frequency of about four times a week, twice a week, once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once every eight weeks, once every twelve weeks, or at a lower frequency, so long as a therapeutic response is achieved.

[0181] In some embodiments, multiple doses of the antibodies disclosed herein are administered to a subject over a defined period of time. In some embodiments, the methods of the present disclosure include sequentially administering multiple doses of an antibody to a subject. As used herein, "sequentially administering" means that each dose of the antibody is administered to the subject at different time points, e.g., on different days separated by a predetermined interval (e.g., hours, days, weeks, or months). In some embodiments, the methods of the present disclosure include sequentially administering a single initial dose of an antibody to a patient, followed by one or more second doses of the antibody, and optionally followed by one or more third doses of the antibody.

[0182] The terms "initial dose," "second dose," and "third dose" refer to the chronological order of administration of the antibody. Thus, an "initial dose" is the dose administered at the start of a treatment regimen (also referred to as a "loading dose"); a "second dose" is the dose administered after the initial dose; and a "third dose" is the dose administered after the second dose. In some embodiments, the initial dose, second dose, and third dose may all contain the same amount of antibody, but may differ from each other in terms of administration frequency. In some embodiments, the amounts of antibody contained in the initial dose, second dose, and / or third dose differ from each other during the course of treatment (e.g., adjusted up or down as appropriate). In certain embodiments, one or more (e.g., 1, 2, 3, 4, or 5) doses are administered at the start of a treatment regimen as a "loading dose," followed by subsequent doses administered at a lower frequency (e.g., a "maintenance dose"). In some embodiments, the initial dose and one or more second doses each contain the same amount of antibody. In other embodiments, the initial dose contains a first amount of antibody, and one or more second doses each contain a second amount of antibody. For example, the first amount of antibody may be 1.5x, 2x, 2.5x, 3x, 3.5x, 4x, or 5x or more of the second amount of antibody.

[0183] In some embodiments, each second dose and / or third dose is administered 1 to 14 weeks (e.g., 1, 1 1 / 2, 2, 2 1 / 2, 3, 3 1 / 2, 4, 4 1 / 2, 5, 5 1 / 2, 6, 6 1 / 2, 7, 7 1 / 2, 8, 8 1 / 2, 9, 9 1 / 2, 10, 10 1 / 2, 11, 11 1 / 2, 12, 12 1 / 2, 13, 13 1 / 2, 14, 14 1 / 2, or longer) after the immediately preceding dose. As used herein, the phrase "immediately preceding dose" means that, in the order of multiple administrations, the dose of the antibody is administered to the patient immediately before the next dose in the sequence, without an intervening dose.

[0184] The methods of the present disclosure can include administering to a patient any number of second and / or third doses of an antibody. For example, in certain embodiments, only a single second dose is administered to the patient. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8 or more) second doses are administered to the patient. Similarly, in certain embodiments, only a single third dose is administered to the patient. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8 or more) third doses are administered to the patient.

[0185] In some embodiments involving multiple second doses, each second dose is administered at the same frequency as the other second doses. For example, each second dose can be administered to the patient 1 week, 2 weeks, 3 weeks, 4 weeks after the immediately preceding dose. Similarly, in some embodiments involving multiple third doses, each third dose is administered at the same frequency as the other third doses. Alternatively, during the course of the treatment regimen, the frequency of the second and / or third doses administered to the patient can vary. During the course of the treatment, the physician can also adjust the administration frequency according to the needs of an individual patient after a clinical examination.

[0186] Combination therapy

[0187] In some embodiments, the antibodies or antigen-binding fragments of the present disclosure can be used in combination with one or more other therapeutic agents. In some embodiments, the additional therapeutic agents are antibodies, small molecules, inhibitory nucleic acids (e.g., RNAi), antibody-drug conjugates, bispecific antibodies, or combinations thereof.

[0188] In some embodiments, the anti-artemin antibodies of the present disclosure can be administered in combination with another artemin inhibitor, an inhibitor of GFRα3 (e.g., the antibody disclosed in WO 2014 / 031712, incorporated herein by reference), an inhibitor of another GDNF family ligand or receptor (e.g., an inhibitor of GDNF, neurturin (NRTN), persephin (PSPN), GFRα1, GFRα2, or GFRα4), or a RET inhibitor.

[0189] In some embodiments, the anti-artemin antibody is administered in combination with additional therapeutic agents that reduce, alleviate, or improve pain. Exemplary additional therapeutic agents include, but are not limited to: COX-2 inhibitors; local anesthetics; NMDA modulators; cannabinoid receptor agonists; P2X family modulators; VR1 antagonists; substance P antagonists; inhibitors of voltage-gated sodium channels (Nav), such as Nav1.7 antagonists, Nav1.8 antagonists, or Nav1.9 antagonists (e.g., antibodies, small molecule inhibitors, or inhibitory nucleic acids directed against Nav1.7, Nav1.8, or Nav1.9); calcium channel inhibitors; potassium channel inhibitors; cytokine inhibitors or cytokine receptor antagonists (e.g., interleukin-1 (IL-1) inhibitors (such as rilonacept (“IL-1trap”); Regeneron) or interleukin-1 receptor (IL-1R) inhibitors, IL-18 inhibitors, IL-6 or IL-6R inhibitors, IL-17 inhibitors, tumor necrosis factor (TNF) or TNF receptor inhibitors (e.g., adalimumab) or inhibitors of TWEAK (TNF-related weak inducer of apoptosis)); growth factor inhibitors (e.g., nerve growth factor (NGF) inhibitors, such as small molecule NGF antagonists or anti-NGF antibodies); neurotrophic factor inhibitors (e.g., inhibitors of BDNF, TrkA, TrkB, or p75, or another inhibitor of GDNF family ligands or receptors); anti-epileptic / anti-convulsant drugs (e.g., gabapentin, pregabalin); opioids; morphine; low-dose colchicine; aspirin or other NSAIDs; steroids (e.g., prednisone, methotrexate, etc.); low-dose cyclosporine A; selective serotonin reuptake inhibitors (SSRI); serotonin-norepinephrine reuptake inhibitors (SNRI); tricyclic antidepressant drugs; inhibitors of acid-sensing ion channels (e.g., ASIC1 or ASIC3); uric acid synthesis inhibitors (e.g., allopurinol); uric acid excretion promoters (e.g., probenecid, sulfinpyrazone, benzbromarone, etc.); and / or corticosteroids.

[0190] The additional therapeutically active component can be administered before, simultaneously with, or shortly after administration of the antibody of the present disclosure. For the purposes of the present disclosure, such administration regimens are considered to be the antibody administered “in combination with” the additional therapeutically active component.

[0191] The present disclosure includes pharmaceutical compositions, wherein the antibodies of the present disclosure are co-formulated with additional therapeutically active components as described elsewhere herein.

[0192] Examples

[0193] The following examples are presented to provide a complete disclosure and description to those of ordinary skill in the art of how to make and use the methods and compositions of the present disclosure and are not intended to limit the scope which the inventors regard as their invention. Efforts have been made to ensure the accuracy of the numbers used (e.g., amounts, temperatures, etc.), but some experimental errors and deviations should be accounted for. Unless otherwise indicated, parts are parts by weight, molecular weights are average molecular weights, temperature is in degrees Celsius, and pressure is at or near atmospheric pressure.

[0194] Example 1: Generation of Human Antibodies Against Human Artemin

[0195] Antibodies against human artemin were obtained by immunizing VELOCIMMUNE mice (i.e., engineered mice containing DNA encoding the variable regions of the human immunoglobulin heavy and κ chains) with the human artemin antigen (hARTN(A108 - G220) with a C-terminal mmH tag; SEQ ID NO:82), and then boosting with the same immunogen.

[0196] After immunization, antibodies were isolated directly from antigen-positive mouse B cells, for example, as described in U.S. Patent No. 7,582,298, which is incorporated herein by reference. Using this method, fully human anti-hARTN antibodies (i.e., antibodies having human variable domains and human constant domains) were obtained. The anti-ARTN antibodies produced using this method were named H4H33331P, H4H33335P, H4H33336P, and H4H33349P. Certain biological properties of exemplary anti-ARTN antibodies produced according to the method of this example are described in detail in the examples shown below.

[0197] Table 1: Amino acid sequence identifiers

[0198]

[0199] Table 2: Nucleic acid sequence identifiers

[0200]

[0201] Example 2: Binding Characteristics of Artemin Antibodies

[0202] Octet binding of Artemin monoclonal antibodies

[0203] Binding of Artemin to each of the Artemin monoclonal antibodies (mAbs) H4H33331P, H4H33335P, H4H33336P, and H4H33349P was determined using a real-time, label-free biolayer interferometry (BLI) assay on the Octet HTX biosensor platform (Pall ForteBio Corp.). Labeled Artemin proteins were generated that comprised the mature region of the following: human (hArtemin, Uniprot Q5T4W7-1, amino acids A108-G220), cynomolgus monkey (Macaca fascicularis, MfArtemin, XP_015292608.1, amino acids A108-G220; X110G, X111P, X112G, X113S, X114R, X115P, X116R; X represents the human sequence at those specific positions) or mouse (mArtemin, Uniprot Q9Z0L2-1, amino acids A112-G224) proteins that were fused at the C-terminus to the epitope tag myc-myc-6xHis. The sequence of the labeled human Artemin, designated “hArtemin-MMH,” is shown in SEQ ID NO:82; the sequence of the labeled cynomolgus monkey Artemin, designated “MfArtemin-MMH,” is shown in SEQ ID NO:83; and the sequence of the labeled mouse Artemin, designated “mArtemin-MMH” or “msArtemin-MMH,” is shown in SEQ ID NO:84.

[0204] The entire experiment was conducted at 25° C. in 10 mM HEPES, 150 mM NaCl, 3 mM EDTA, 0.02% NaN3 and 0.05% v / v surfactant Tween-20, 1 mg / mL BSA, pH 7.4 (HBS-EBT) buffer, where the plate was shaken at 1000 rpm. Different Artemin mAbs were first captured onto anti-human antibody (AHC)-coated Octet biosensor tips (Fortebio Inc, #18-5064) by immersing the biosensor tips in wells containing 2 μg / mL Artemin mAb for 40 seconds. Subsequently, the Artemin mAb-captured biosensor tips were immersed in wells containing different concentrations (30-3.75 nM, 2-fold serial dilutions) of labeled human, cynomolgus monkey, or mouse Artemin proteins for 3 minutes, and their dissociation in HBS-EBT buffer was monitored for 20 minutes. All Artemin and Artemin mAb samples were prepared in HBS-EBT buffer. The real-time binding reactions were monitored throughout the experiment, and the binding reactions at the end of each step were recorded.

[0205] The binding of various Artemin reagents to different Artemin mAbs is reported in Table 3, and the concentration-dependent binding data are also provided in Figures 1 to 3 .

[0206] As shown in Table 3 and Figure 1 , all four Artemin mAbs bind hArtemin-MMH at 25 °C.

[0207] As shown in Table 3 and Figure 2 , all four Artemin mAbs bind MfArtemin-MMH at 25 °C.

[0208] As shown in Table 3 and Figure 3 , all four Artemin mAbs bind mArtemin-MMH at 25 °C.

[0209] Table 3: Various Artemin reagents and different Artemin Binding of mAb

[0210]

[0211] $ Indicates that at the indicated concentrations, the binding of Artemin to the surface captured by the isotype mAb was not tested.

[0212] Biacore binding kinetics of Artemin monoclonal antibodies

[0213] The equilibrium dissociation constants (K D)。All binding studies were performed at 25 °C in 10 mM HEPES, 150 mM NaCl, 3 mM EDTA, and 0.05% v / v surfactant Tween-20 (pH 7.4) (HBS-ET) running buffer. The surface of the Biacore CM4 sensor chip was first derivatized by amine coupling of anti-myc mAb to capture hArtemin-MMH, MfArtemin-MMH, or mArtemin-MMH. Different concentrations of Artemin mAb (60 - 2.2 nM, 3-fold serial dilution) prepared in HBS-EP running buffer were injected at a flow rate of 50 μL / min onto the Artemin-captured surface for 3 minutes, and their dissociation in HBS-ET running buffer was monitored for 5 minutes. At the end of each cycle, the Artemin-captured surface was regenerated using a 6-second injection of 20 mM phosphoric acid.

[0214] The real-time binding sensorgrams were fitted to a 1:1 binding model with mass transfer limitation using Scrubber 2.0c curve fitting software to determine the association rate (k a ) and dissociation rate (k d ). The binding dissociation equilibrium constant (K D ) and dissociation half-life (t1 / 2) were calculated from the kinetic rates as follows: K D (M) = k d / k a , and t 1 / 2 (min) = [ln2 / (60 * k d )].

[0215] The binding kinetic parameters of Artemin binding to different Artemin mAbs of the present invention at 25 °C are shown in Tables 4 - 6.

[0216] As shown in Table 4, all four Artemin mAbs bind hArtemin-MMH with K D values in the range of 119 pM to 2.91 nM at 25 °C.

[0217] As shown in Table 5, three of the four Artemin mAbs (H4H33331P, H4H33335P, and H4H33349P) bind MfArtemin-MMH with K D values in the range of 111 pM to 1.51 nM at 25 °C. Specific binding was also observed for mAb H4H33335P, but the binding kinetic parameters could not be determined under the current experimental conditions.

[0218] As shown in Table 6, two of the four Artemin mAbs (H4H33331P and H4H33349P) bound to mArtemin-MMH with K D values of 113 pM and 346 pM, respectively, at 25°C. Specific binding was also observed for mAb H4H33335P and H4H33336P, but the binding kinetic parameters could not be determined under the current experimental conditions.

[0219] Table 4: Binding kinetic parameters of different Artemin monoclonal antibodies to hArtemin-MMH at 25°C

[0220]

[0221] * Indicates unbound (NB) observed under the current experimental conditions.

[0222] Table 5: Binding kinetic parameters of different Artemin monoclonal antibodies to MfArtemin-MMH at 25°C

[0223]

[0224] * Indicates unbound (NB) observed under the current experimental conditions.

[0225] # Indicates that specific binding was observed, but the binding kinetic parameters could not be determined under the current experimental conditions and are therefore not available (NA).

[0226] Table 6: Binding kinetic parameters of different Artemin monoclonal antibodies to mArtemin-MMH at 25°C

[0227]

[0228] * Indicates unbound (NB) observed under the current experimental conditions.

[0229] # Indicates that specific binding was observed, but the binding kinetic parameters could not be determined under the current experimental conditions and are therefore not available (NA).

[0230] Example 3: Anti-ARTN Antibody Blocks the Binding of ARTN to GFRα3

[0231] Multiple artemin blocking assays were developed using an ELISA-based format to determine the ability of anti-artemin (ARTN) antibodies to block the binding of monomeric human, monkey (cynomolgus), and mouse ARTN proteins to human GFRα3 (glial cell line-derived neurotrophic factor (GDNF) family receptor α3).

[0232] The Artemin proteins used in the experiments were hArtemin-MMH, MfArtemin-MMH, or mArtemin-MMH as disclosed in Example 2 above. The human GFRα3 used in the experiments consisted of UniProtKB accession number O60609, amino acids D32-W382, with the Fc portion of human IgG1 at the C-terminus, and was designated "hGFRa3.hFc" (SEQ ID NO:85).

[0233] The experiments were conducted using the following procedure. For all three assays, the plates were coated overnight at 4 °C in PBS with hGFRa3.hFc at 2 μg / ml on 96-well microtiter plates. After overnight coating, the non-specific binding sites were then blocked at room temperature for one hour using PBS containing 0.5% (w / v) BSA. In other microtiter plates, a constant amount of 75 pM hARTN.mmh, 400 pM mfARTN.mmh, or 150 pM msARTN.mmh was respectively combined with anti-ARTN antibodies and an irrelevant IgG4 antibody control in PBS + 0.5% BSA in a serial dilution from 0.0017 nM to 100 nM. After incubation for one hour, the mixture solution was transferred to the microtiter plates coated with hGFRa3.hFc. After incubation for one hour at RT, the wells were washed with PBST, and the plate-bound ARTN was detected using an anti-penta-His antibody conjugated to horseradish peroxidase (HRP) (Qiagen). Then, the plates were developed using a TMB substrate solution (BD Biosciences, #555214) according to the manufacturer's recommendations, and the absorbance at 450 nm was measured on a Victor X5 plate reader.

[0234] Using Prism TM software (GraphPad), the binding data were analyzed using a sigmoidal dose-response model. The calculated IC50 value, defined as the antibody concentration required to block 50% of the binding of ARTN.mmh to the plate-coated hGFRa3.hFc, was used as an indicator of blocking potency. Based on the binding stoichiometry of two artemins to one antibody, the lower limit of quantification (LLOA) for the IC50 of hARTN.mmh, mfARTN.mmh, and mARTN.mmh was 18.8 pM, 100 pM, and 37.5 pM, respectively. If the calculated IC50 value was below the LLOA, less than the LLOA was reported. Based on the background-corrected binding signal observed at the highest antibody concentration tested, the percent blocking of the 100 nM antibody was calculated using the formula listed below, and all tested antibodies were reported:

[0235]

[0236] Results

[0237] The ability of anti-ARTN antibodies to block the binding of human, monkey, and mouse artemin to human GFRa3 was evaluated using multiple blocking ELISA formats. In these assays, various concentrations of anti-ARTN antibodies were titrated against 75 pM hARTN.mmh, 400 pM mfARTN.mmh, or 150 pM msARTN.mmh, and the binding of the three artemins to hGFRa3.hFc in the presence of the antibodies was evaluated. Plate-bound ARTN.mmh was detected using an HRP-conjugated anti-penta-His antibody.

[0238] Four test antibodies effectively blocked the binding of human, monkey, and mouse artemin to plate-bound human GFRa3, with IC50 values below 1 nM and blocking percentages greater than 50% at the highest antibody concentration tested. Table 7 summarizes the blocking IC50 and blocking percentages at the highest tested concentration of anti-ARTN antibodies. Unrelated isotype control antibodies did not exhibit blocking activity.

[0239] Table 7: Potency of antibodies blocking human, monkey and mouse Artemin binding to human GFRα3

[0240]

[0241] *Below the limit of detection

[0242] Example 4: Inhibition of Artemin-mediated cell signaling by Artemin monoclonal antibodies

[0243] To evaluate the inhibition of Artemin-mediated cell signaling by anti-Artemin antibodies, a biological assay was established in HEK293 cells (human embryonic kidney, ATCC, #CRL-1573). HEK293 cells were transduced with a serum response element luciferase reporter gene (SRE-Luc, SA Biosciences) and engineered to stably express full-length human GFRα3 (amino acids M1 to W400, GenBank accession number NP_001487.2) and full-length human RET isoform c (amino acids M1 to F1073, GenBank accession number NP_065681.1, with two amino acid substitutions at G691S and R982C). Optimal cell clones were isolated and the resulting cell line was named HEK293 / SRELuc / hRET / hGFRa3 cl.A3. The Artemin proteins hArtemin-MMH, MfArtemin-MMH, or mArtemin-MMH were disclosed in Example 2 above.

[0244] For the biological assay, cells were seeded at 2.5×10 in assay buffer (0.1% FBS in Opti-MEM medium)4 Cells were seeded at TM a density of 1 cell / well into a 96-well plate and cultured overnight (37 °C, 5% CO2). The next day, the antibodies of the present invention or isotype control antibodies were serially diluted (1:3) in assay medium to a final concentration of 100 nM to 1.7 pM (with additional wells for a separate assay medium without the test molecule), and added to the cells containing a fixed concentration of Artemin (500 pM hArtemin, 3 nM mfArtemin, or 1 nM mArtemin) at 37 °C in 5% CO2. To assess the degree of signaling activation, Artemin (h, mf, or m) was serially diluted (1:3) to a final concentration of 100 nM to 1.7 pM (with additional wells for a separate assay medium) and added to the cells. After a 5-hour incubation, luciferase activity was evaluated by adding OneGlo

[0245] Luciferase Assay System reagent (Promega E6130), and relative light units (RLU) were measured using an Envision multimode plate reader (PerkinElmer). The results were analyzed using nonlinear regression (4-parameter logistic) in Artemin GraphPad Prism 8 to obtain EC50 and IC50 values. The percent inhibition was calculated using the following equation: inhibition % Inhibition = 100 × [(RLU Artemin – RLU Baseline ) / (RLU

[0246] In this equation, “RLU Artemin ” refers to the RLU value of cells treated with constant Artemin without antibody. “RLU inhibition ” refers to the RLU value measured at the maximum concentration of antibody with constant Artemin. “RLU Baseline ” refers to the RLU value of cells measured in the absence of ligand or antibody.

[0247] As shown in Table 8, the anti-Artemin antibodies H4H33331P, H4H33335P, H4H33336P, and H4H33349P were tested on HEK293 / SRELuc / hRET / hGFRa3 cl.A3 cells with 500 pM hArtemin, 3 nM mfArtemin, or 1 nM mArtemin. Human, macaque, and mouse Artemin activate signaling through the human GFRα3 and Ret receptors, with EC50 values of 813 pM, 2.26 nM, and 620 pM, respectively.

[0248] Table 8: Inhibition of human, macaqueMonkey and mouse Artemin by anti-Artemin antibodies in HEK293 / SRELuc / hRET / hGFRa3 cI.A3 cells

[0249]

[0250] Example 5: Inhibition of plantar Artemin-induced thermal hyperalgesia by Artemin monoclonal antibody

[0251] Animal subjects

[0252] Adult C57BL / 6 male mice (Jackson Laboratories) at 10 - 16 weeks of age were used for the experiments. The animals were housed in groups of up to five in a temperature-controlled environment on a 12-hour light / dark cycle with free access to water and standard laboratory chow. All experimental procedures were approved by the Regeneron Animal Care and Use Committee and were conducted by an experimenter blinded to the animal groupings.

[0253] Adaptation and baseline

[0254] One week prior to plantar injection, the mice were placed in the Hargreaves behavioral test apparatus for at least two hours per day to acclimate them to the room and the apparatus. Prior to each test, the animals were additionally acclimated for at least one hour. Baseline nociceptive responses were performed on the weekend.

[0255] Induction of growth factor-induced plantar thermal hyperalgesia

[0256] Prior to subcutaneous (s.c.) injection of 10 mg / kg of artemin antibody, baseline thermal hyperalgesia was evaluated using the Hargreaves test (described below). Three days later, by placing the mice in a transparent plastic restraint and exposing their hindlimbs, PBS (one group) or 20 μl of saline containing recombinant murine artemin from R&D Systems (five groups) was injected into the hindpaw. The left hindlimb was extended and 0.5 μg artemin was injected onto the plantar surface of the paw. Thermal hyperalgesia was evaluated again 4 days later. Using the results of the artemin injection trials at a power level of.80 and an α of.05, it was calculated that n of 7 - 8 animals would be required to detect a return to baseline latency to stop antibody treatment. A sample size of 7 animals was chosen for the experiment because it was expected that the growth factor response would be completely blocked, and thus the animal usage could be minimized by selecting the smallest recommended sample size from the power analysis.

[0257] Thermal hyperalgesia: The thermal sensitivity of animals was tested using the Hargreaves test (Hargreaves et al., Pain, 1988, 32:77 - 88). Mice were placed on a 30 °C heated glass in each chamber (IITC model 390G with heating base model 400) and allowed to acclimate to the environment before readings. The device was set to a laser intensity of 15%, and testing was performed using repeated measures (three measurements per foot) on the plantar surface of the ipsilateral hind paw. The mean latency to withdrawal was recorded, allowing a maximum beam exposure of 20 seconds to avoid tissue damage. One-way independent group ANOVA was used to statistically analyze the withdrawal latency to compare treatment groups, with α set at 0.05. Tukey's post hoc test was used to explore significant main effects of the treatment.

[0258] Results

[0259] Four selected human anti-artemin antibodies (H4H33331P, H4H33335P, H4H33336P, and H4H33349P) were evaluated in a model of plantar nerve growth factor-induced hyperalgesia. As Figure 4 shown, in the Hargreaves test with subcutaneous injection at 10 mg / kg, all four drugs showed complete blockade of artemin-induced thermal hyperalgesia, such that the withdrawal latency was similar to that of the group treated with PBS in the plantar nerve. The withdrawal thresholds of animals receiving anti-artemin antibodies were significantly different from those of animals receiving isotype control antibodies, indicating that all four artemin antibodies effectively inhibited artemin-induced hyperalgesia.

[0260]

[0261]

[0262]

[0263]

[0264]

[0265]

[0266]

[0267]

[0268]

[0269]

[0270] The present invention is not limited in scope by the specific embodiments described herein. In fact, various modifications of the invention will be apparent to those skilled in the art in addition to those described herein, from the foregoing description and drawings. Such modifications are intended to fall within the scope of the appended claims. The disclosures of all patents and non-patent documents cited herein are hereby incorporated by reference in their entirety. Sequence Listing <110> Regeneron Pharmaceuticals, Inc. CROLL, Susan D. MACDONALD, Lynn MURPHY, Andrew J. <120> Human Antibodies Against Artemin and Methods of Use Thereof <130> 10907WO01 <140> TBD <141> 2022-04-19 <150> 63 / 177,369 <151> 2021-04-20 <160> 85 <170> PatentIn version 3.5 <210> 1 <211> 366 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 1 gaggtgcagc tggtggagag cggcggcgac ctggtgcagc ccggcggcag cctgaagctg 60 agctgcgccg ccagcggctt caccttcagc ggcagcgcca tgcactgggt gaggcaggcc 120 agcggcaagg gcctggagtg ggtgggcagg atcaggaaca aggtgcacag gtacgccacc 180 gagtacggcg ccagcgtgaa gggcaggttc accatcagca gggacgacag caagaacacc 240 gcctacctgc agatgaacag cctgaagatc gaggacaccg ccgtgtacta ctgcgtggtg 300 gtggtgcccg gcagcatcga ggccttcgac atctggggcc agggcaccat ggtgaccgtg 360 agcagc 366 <210> 2 <211> 122 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 2 Glu Val Gln Leu Val Glu Ser Gly Gly Asp Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Gly Ser 20 25 30 Ala Met His Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Arg Asn Lys Val His Arg Tyr Ala Thr Glu Tyr Gly Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Ala Tyr Leu Gln Met Asn Ser Leu Lys Ile Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Val Val Val Val Pro Gly Ser Ile Glu Ala Phe Asp Ile Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 <210> 3 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 3 ggcttcacct tcagcggcag cgcc 24 <210> 4 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 4 Gly Phe Thr Phe Ser Gly Ser Ala 1 5 <210> 5 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 5 atcaggaaca aggtgcacag gtacgccacc 30 <210> 6 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 6 Ile Arg Asn Lys Val His Arg Tyr Ala Thr 1 5 10 <210> 7 <211> 39 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 7 gtggtggtgg tgcccggcag catcgaggcc ttcgacatc 39 <210> 8 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 8 Val Val Val Val Pro Gly Ser Ile Glu Ala Phe Asp Ile 1 5 10 <210> 9 <211> 321 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 9 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagggtgacc 60 atcacctgca gggccagcca gagcatcagc acctacctga actggtacca gcagaagccc 120 ggcaaggccc ccaacctgct gatctacgcc gccagcaccc tggagagcgg cgtgcccagc 180 aggttcagcg gcagcggcag cggcaccgac ttcaccctga ccatcagcag cctgcagccc 240 gaggacttcg agatctacta ctgccagcag agctacaaca tcccctggac cttcggccag 300 ggcaccaagg tggagatcaa g 321 <210> 10 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 10 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Thr Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Glu Ile Tyr Tyr Cys Gln Gln Ser Tyr Asn Ile Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 11 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 11 cagagcatca gcacctac 18 <210> 12 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 12 Gln Ser Ile Ser Thr Tyr 1 5 <210> 13 <211> 9 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 13 gccgccagc 9 <210> 14 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 14 Ala Ala Ser 1 <210> 15 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> Synthesis <400> 15 cagcagagct acaacatccc ctggacc 27 <210> 16 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Synthesis <400> 16 Gln Gln Ser Tyr Asn Ile Pro Trp Thr 1 5 <210> 17 <211> 1347 <212> DNA <213> Artificial Sequence <220> <223> Synthesis <400> 17 gaggtgcagc tggtggagag cggcggcgac ctggtgcagc ccggcggcag cctgaagctg 60 agctgcgccg ccagcggctt caccttcagc ggcagcgcca tgcactgggt gaggcaggcc 120 agcggcaagg gcctggagtg ggtgggcagg atcaggaaca aggtgcacag gtacgccacc 180 gagtacggcg ccagcgtgaa gggcaggttc accatcagca gggacgacag caagaacacc 240 gcctacctgc agatgaacag cctgaagatc gaggacaccg ccgtgtacta ctgcgtggtg 300 gtggtgcccg gcagcatcga ggccttcgac atctggggcc agggcaccat ggtgaccgtg 360 agcagcgcca gcaccaaggg ccccagcgtg ttccccctgg ccccctgcag caggagcacc 420 agcgagagca ccgccgccct gggctgcctg gtgaaggact acttccccga gcccgtgacc 480 gtgagctgga acagcggcgc cctgaccagc ggcgtgcaca ccttccccgc cgtgctgcag 540 agcagcggcc tgtacagcct gagcagcgtg gtgaccgtgc ccagcagcag cctgggcacc 600 aagacctaca cctgcaacgt ggaccacaag cccagcaaca ccaaggtgga caagagggtg 660 gagagcaagt acggcccccc ctgccccccc tgccccgccc ccgagttcct gggcggcccc 720 agcgtgttcc tgttcccccc caagcccaag gacaccctga tgatcagcag gacccccgag 780 gtgacctgcg tggtggtgga cgtgagccag gaggaccccg aggtgcagtt caactggtac 840 gtggacggcg tggaggtgca caacgccaag accaagccca gggaggagca gttcaacagc 900 acctacaggg tggtgagcgt gctgaccgtg ctgcaccagg actggctgaa cggcaaggag 960 tacaagtgca aggtgagcaa caagggcctg cccagcagca tcgagaagac catcagcaag 1020 gccaagggcc agcccaggga gccccaggtg tacaccctgc cccccagcca ggaggagatg 1080 accaagaacc aggtgagcct gacctgcctg gtgaagggct tctaccccag cgacatcgcc 1140 gtggagtggg agagcaacgg ccagcccgag aacaactaca agaccacccc ccccgtgctg 1200 gacagcgacg gcagcttctt cctgtacagc aggctgaccg tggacaagag caggtggcag 1260 gagggcaacg tgttcagctg cagcgtgatg cacgaggccc tgcacaacca ctacacccag 1320 aagagcctga gcctgagcct gggcaag 1347 <210> 18 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 18 Glu Val Gln Leu Val Glu Ser Gly Gly Asp Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Gly Ser 20 25 30 Ala Met His Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Arg Asn Lys Val His Arg Tyr Ala Thr Glu Tyr Gly Ala 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Ala Tyr Leu Gln Met Asn Ser Leu Lys Ile Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Val Val Val Val Pro Gly Ser Ile Glu Ala Phe Asp Ile Trp 100 105 110 Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr 210 215 220 Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp 260 265 270 Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly 435 440 445 Lys <210> 19 <211> 642 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 19 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagggtgacc 60 atcacctgca gggccagcca gagcatcagc acctacctga actggtacca gcagaagccc 120 ggcaaggccc ccaacctgct gatctacgcc gccagcaccc tggagagcgg cgtgcccagc 180 aggttcagcg gcagcggcag cggcaccgac ttcaccctga ccatcagcag cctgcagccc 240 gaggacttcg agatctacta ctgccagcag agctacaaca tcccctggac cttcggccag 300 ggcaccaagg tggagatcaa gaggaccgtg gccgccccca gcgtgttcat cttccccccc 360 agcgacgagc agctgaagag cggcaccgcc agcgtggtgt gcctgctgaa caacttctac 420 cccagggagg ccaaggtgca gtggaaggtg gacaacgccc tgcagagcgg caacagccag 480 gagagcgtga ccgagcagga cagcaaggac agcacctaca gcctgagcag caccctgacc 540 ctgagcaagg ccgactacga gaagcacaag gtgtacgcct gcgaggtgac ccaccagggc 600 ctgagcagcc ccgtgaccaa gagcttcaac aggggcgagt gc 642 <210> 20 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 20 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Thr Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Glu Ile Tyr Tyr Cys Gln Gln Ser Tyr Asn Ile Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 21 <211> 360 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 21 gaggtgcagc tggtggagag cggcggcggc ctggtgcagc ccggcggcag cctgaggctg 60 agctgcgccg ccagcggctt caccttcagc agctacgaca tgcactgggt gaggcaggcc 120 accggcaagg gcctggagtg ggtgagcacc atcgacaccg ccggcgacac ctactacccc 180 ggcagcgtga agggcaggtt caccatcagc agggagaacg ccaagaacag cctgtacctg 240 cagatgaaca gcctgagggc cggcgacacc gccgtgtact actgcgccag ggacggcgag 300 ctggagctgc agggctactt cgacctgtgg ggcaggggca ccctggtgac cgtgagcagc 360 <210> 22 <211> 120 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 22 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Thr Ile Asp Thr Ala Gly Asp Thr Tyr Tyr Pro Gly Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Asp Gly Glu Leu Glu Leu Gln Gly Tyr Phe Asp Leu Trp Gly Arg 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 23 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 23 ggcttcacct tcagcagcta cgac 24 <210> 24 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 24 Gly Phe Thr Phe Ser Ser Tyr Asp 1 5 <210> 25 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 25 atcgacaccg ccggcgacac c 21 <210> 26 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 26 Ile Asp Thr Ala Gly Asp Thr 1 5 <210> 27 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 27 gccagggacg gcgagctgga gctgcagggc tacttcgacc tg 42 <210> 28 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 28 Ala Arg Asp Gly Glu Leu Glu Leu Gln Gly Tyr Phe Asp Leu 1 5 10 <210> 29 <211> 321 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 29 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagggtgacc 60 atcacctgca gggccagcca gatcatcagc atctacctga actggtacca gcagaagccc 120 ggcaaggccc ccaagctgct gatctacgcc gccagcagcc tgcagagcgg cgtgcccagc 180 aggttcagcg gcagcggcag cggcaccgac ttcaccctga ccatcagcag cctgcagccc 240 gaggacttcg ccacctacta ctgccagcag agctacacca cccccctgac cttcggcggc 300 ggcaccaagg tggagatcaa g 321 <210> 30 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 30 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ile Ile Ser Ile Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Thr Thr Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 31 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 31 cagatcatca gcatctac 18 <210> 32 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 32 Gln Ile Ile Ser Ile Tyr 1 5 <210> 33 <211> 9 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 33 gccgccagc 9 <210> 34 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 34 Ala Ala Ser 1 <210> 35 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 35 cagcagagct acaccacccc cctgacc 27 <210> 36 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 36 Gln Gln Ser Tyr Thr Thr Pro Leu Thr 1 5 <210> 37 <211> 1341 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 37 gaggtgcagc tggtggagag cggcggcggc ctggtgcagc ccggcggcag cctgaggctg 60 agctgcgccg ccagcggctt caccttcagc agctacgaca tgcactgggt gaggcaggcc 120 accggcaagg gcctggagtg ggtgagcacc atcgacaccg ccggcgacac ctactacccc 180 ggcagcgtga agggcaggtt caccatcagc agggagaacg ccaagaacag cctgtacctg 240 cagatgaaca gcctgagggc cggcgacacc gccgtgtact actgcgccag ggacggcgag 300 ctggagctgc agggctactt cgacctgtgg ggcaggggca ccctggtgac cgtgagcagc 360 gccagcacca agggccccag cgtgttcccc ctggccccct gcagcaggag caccagcgag 420 agcaccgccg ccctgggctg cctggtgaag gactacttcc ccgagcccgt gaccgtgagc 480 tggaacagcg gcgccctgac cagcggcgtg cacaccttcc ccgccgtgct gcagagcagc 540 ggcctgtaca gcctgagcag cgtggtgacc gtgcccagca gcagcctggg caccaagacc 600 tacacctgca acgtggacca caagcccagc aacaccaagg tggacaagag ggtggagagc 660 aagtacggcc ccccctgccc cccctgcccc gcccccgagt tcctgggcgg ccccagcgtg 720 ttcctgttcc cccccaagcc caaggacacc ctgatgatca gcaggacccc cgaggtgacc 780 tgcgtggtgg tggacgtgag ccaggaggac cccgaggtgc agttcaactg gtacgtggac 840 ggcgtggagg tgcacaacgc caagaccaag cccagggagg agcagttcaa cagcacctac 900 agggtggtga gcgtgctgac cgtgctgcac caggactggc tgaacggcaa ggagtacaag 960 tgcaaggtga gcaacaaggg cctgcccagc agcatcgaga agaccatcag caaggccaag 1020 ggccagccca gggagcccca ggtgtacacc ctgcccccca gccaggagga gatgaccaag 1080 aaccaggtga gcctgacctg cctggtgaag ggcttctacc ccagcgacat cgccgtggag 1140 tgggagagca acggccagcc cgagaacaac tacaagacca ccccccccgt gctggacagc 1200 gacggcagct tcttcctgta cagcaggctg accgtggaca agagcaggtg gcaggagggc 1260 aacgtgttca gctgcagcgt gatgcacgag gccctgcaca accactacac ccagaagagc 1320 ctgagcctga gcctgggcaa g 1341 <210> 38 <211> 447 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 38 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Thr Ile Asp Thr Ala Gly Asp Thr Tyr Tyr Pro Gly Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Asp Gly Glu Leu Glu Leu Gln Gly Tyr Phe Asp Leu Trp Gly Arg 100 105 110 Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro 210 215 220 Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val 225 230 235 240 Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255 Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu 260 265 270 Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275 280 285 Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser 290 295 300 Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys 305 310 315 320 Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile 325 330 335 Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 340 345 350 Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 355 360 365 Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 370 375 380 Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 385 390 395 400 Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg 405 410 415 Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 420 425 430 His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys 435 440 445 <210> 39 <211> 642 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 39 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagggtgacc 60 atcacctgca gggccagcca gatcatcagc atctacctga actggtacca gcagaagccc 120 ggcaaggccc ccaagctgct gatctacgcc gccagcagcc tgcagagcgg cgtgcccagc 180 aggttcagcg gcagcggcag cggcaccgac ttcaccctga ccatcagcag cctgcagccc 240 gaggacttcg ccacctacta ctgccagcag agctacacca cccccctgac cttcggcggc 300 ggcaccaagg tggagatcaa gaggaccgtg gccgccccca gcgtgttcat cttccccccc 360 agcgacgagc agctgaagag cggcaccgcc agcgtggtgt gcctgctgaa caacttctac 420 cccagggagg ccaaggtgca gtggaaggtg gacaacgccc tgcagagcgg caacagccag 480 gagagcgtga ccgagcagga cagcaaggac agcacctaca gcctgagcag caccctgacc 540 ctgagcaagg ccgactacga gaagcacaag gtgtacgcct gcgaggtgac ccaccagggc 600 ctgagcagcc ccgtgaccaa gagcttcaac aggggcgagt gc 642 <210> 40 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 40 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ile Ile Ser Ile Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Thr Thr Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 41 <211> 372 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 41 caggtgcagc tggtggagag cggcggcggc gtggtgcagc ccggcaggag cctgaggctg 60 agctgcgtgg ccagcggctt catcctgagc cactacggca tgcactgggt gaggcaggcc 120 cccggcaagg gcctggagtg ggtggccgtg atctggttcg acggcaccaa caagtactac 180 gccgacagcg tgaagggcag gttcaccgtg agcagggaca acagcaagaa caccctgtac 240 atgcagatga acagcctgag ggccgaggac accgccgtgt actactgcgc cagggacccc 300 ccccccgcca ggaggggcaa ctacaacggc atggacgtgt ggggccaggg caccaccgtg 360 accgtgagca gc 372 <210> 42 <211> 124 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 42 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Ile Leu Ser His Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Phe Asp Gly Thr Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Met Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Pro Pro Pro Ala Arg Arg Gly Asn Tyr Asn Gly Met Asp 100 105 110 Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 43 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 43 ggcttcatcc tgagccacta cggc 24 <210> 44 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 44 Gly Phe Ile Leu Ser His Tyr Gly 1 5 <210> 45 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 45 atctggttcg acggcaccaa caag 24 <210> 46 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 46 Ile Trp Phe Asp Gly Thr Asn Lys 1 5 <210> 47 <211> 51 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 47 gccagggacc ccccccccgc caggaggggc aactacaacg gcatggacgt g 51 <210> 48 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 48 Ala Arg Asp Pro Pro Pro Ala Arg Arg Gly Asn Tyr Asn Gly Met Asp 1 5 10 15 Val <210> 49 <211> 321 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 49 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagggtgacc 60 atcacctgca gggccagcca ggacatcagg aacgacctgg gctggtacca gcagaagccc 120 ggcaaggccc ccaagaggct gatcttcggc gccagcagcc tgcagagcgg cgtgcccctg 180 aggttcagcg gcagcggcag cggcaccgag ttcaccctga ccatcaacaa cctgcagccc 240 gaggacttcg ccaccttcta ctgcctgcag gacaacagct acccctggac cttcggccag 300 ggcaccaagg tggagatcaa g 321 <210> 50 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 50 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Arg Asn Asp 20 25 30 Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45 Phe Gly Ala Ser Ser Leu Gln Ser Gly Val Pro Leu Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Asn Asn Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Phe Tyr Cys Leu Gln Asp Asn Ser Tyr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 51 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 51 caggacatca ggaacgac 18 <210> 52 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 52 Gln Asp Ile Arg Asn Asp 1 5 <210> 53 <211> 9 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 53 ggcgccagc 9 <210> 54 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 54 Gly Ala Ser 1 <210> 55 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 55 ctgcaggaca acagctaccc ctggacc 27 <210> 56 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 56 Leu Gln Asp Asn Ser Tyr Pro Trp Thr 1 5 <210> 57 <211> 1353 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 57 caggtgcagc tggtggagag cggcggcggc gtggtgcagc ccggcaggag cctgaggctg 60 agctgcgtgg ccagcggctt catcctgagc cactacggca tgcactgggt gaggcaggcc 120 cccggcaagg gcctggagtg ggtggccgtg atctggttcg acggcaccaa caagtactac 180 gccgacagcg tgaagggcag gttcaccgtg agcagggaca acagcaagaa caccctgtac 240 atgcagatga acagcctgag ggccgaggac accgccgtgt actactgcgc cagggacccc 300 ccccccgcca ggaggggcaa ctacaacggc atggacgtgt ggggccaggg caccaccgtg 360 accgtgagca gcgccagcac caagggcccc agcgtgttcc ccctggcccc ctgcagcagg 420 agcaccagcg agagcaccgc cgccctgggc tgcctggtga aggactactt ccccgagccc 480 gtgaccgtga gctggaacag cggcgccctg accagcggcg tgcacacctt ccccgccgtg 540 ctgcagagca gcggcctgta cagcctgagc agcgtggtga ccgtgcccag cagcagcctg 600 ggcaccaaga cctacacctg caacgtggac cacaagccca gcaacaccaa ggtggacaag 660 agggtggaga gcaagtacgg ccccccctgc cccccctgcc ccgcccccga gttcctgggc 720 ggccccagcg tgttcctgtt cccccccaag cccaaggaca ccctgatgat cagcaggacc 780 cccgaggtga cctgcgtggt ggtggacgtg agccaggagg accccgaggt gcagttcaac 840 tggtacgtgg acggcgtgga ggtgcacaac gccaagacca agcccaggga ggagcagttc 900 aacagcacct acagggtggt gagcgtgctg accgtgctgc accaggactg gctgaacggc 960 aaggagtaca agtgcaaggt gagcaacaag ggcctgccca gcagcatcga gaagaccatc 1020 agcaaggcca agggccagcc cagggagccc caggtgtaca ccctgccccc cagccaggag 1080 gagatgacca agaaccaggt gagcctgacc tgcctggtga agggcttcta ccccagcgac 1140 atcgccgtgg agtgggagag caacggccag cccgagaaca actacaagac cacccccccc 1200 gtgctggaca gcgacggcag cttcttcctg tacagcaggc tgaccgtgga caagagcagg 1260 tggcaggagg gcaacgtgtt cagctgcagc gtgatgcacg aggccctgca caaccactac 1320 acccagaaga gcctgagcct gagcctgggc aag 1353 <210> 58 <211> 451 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 58 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Ile Leu Ser His Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Phe Asp Gly Thr Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Met Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Pro Pro Pro Ala Arg Arg Gly Asn Tyr Asn Gly Met Asp 100 105 110 Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125 Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu 130 135 140 Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro 145 150 155 160 Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr 165 170 175 Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val 180 185 190 Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn 195 200 205 Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser 210 215 220 Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly 225 230 235 240 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln 260 265 270 Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 305 310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445 Leu Gly Lys 450 <210> 59 <211> 642 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 59 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagggtgacc 60 atcacctgca gggccagcca ggacatcagg aacgacctgg gctggtacca gcagaagccc 120 ggcaaggccc ccaagaggct gatcttcggc gccagcagcc tgcagagcgg cgtgcccctg 180 aggttcagcg gcagcggcag cggcaccgag ttcaccctga ccatcaacaa cctgcagccc 240 gaggacttcg ccaccttcta ctgcctgcag gacaacagct acccctggac cttcggccag 300 ggcaccaagg tggagatcaa gaggaccgtg gccgccccca gcgtgttcat cttccccccc 360 agcgacgagc agctgaagag cggcaccgcc agcgtggtgt gcctgctgaa caacttctac 420 cccagggagg ccaaggtgca gtggaaggtg gacaacgccc tgcagagcgg caacagccag 480 gagagcgtga ccgagcagga cagcaaggac agcacctaca gcctgagcag caccctgacc 540 ctgagcaagg ccgactacga gaagcacaag gtgtacgcct gcgaggtgac ccaccagggc 600 ctgagcagcc ccgtgaccaa gagcttcaac aggggcgagt gc 642 <210> 60 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 60 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Arg Asn Asp 20 25 30 Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45 Phe Gly Ala Ser Ser Leu Gln Ser Gly Val Pro Leu Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Asn Asn Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Phe Tyr Cys Leu Gln Asp Asn Ser Tyr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 61 <211> 369 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 61 caggtgcagc tggtggagag cggcggcggc gtggtgcagc ccggcaggag cctgaggctg 60 agctgcgagg ccagcggctt caccttcagc gactacgacc tgcactgggt gaggcaggcc 120 cccggcaagg gcctggagtg ggtggccttc atcagcaacg acggcagcaa cgagtactac 180 cccaagagcg tgaagggcag gttcagcatc agcagggaca acagcaagaa caccctgtac 240 ctgcagatga acagcctgag ggccgaggac accgccgtgt accactgcac caaggagagg 300 gactactact acgacagcag cggcagcctg gactactggg gccagggcac cctggtgacc 360 gtgagcagc 369 <210> 62 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 62 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Asp Leu His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Phe Ile Ser Asn Asp Gly Ser Asn Glu Tyr Tyr Pro Lys Ser Val 50 55 60 Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr His Cys 85 90 95 Thr Lys Glu Arg Asp Tyr Tyr Tyr Asp Ser Ser Gly Ser Leu Asp Tyr 100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 63 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 63 ggcttcacct tcagcgacta cgac 24 <210> 64 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 64 Gly Phe Thr Phe Ser Asp Tyr Asp 1 5 <210> 65 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 65 atcagcaacg acggcagcaa cgag 24 <210> 66 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 66 Ile Ser Asn Asp Gly Ser Asn Glu 1 5 <210> 67 <211> 48 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 67 accaaggaga gggactacta ctacgacagc agcggcagcc tggactac 48 <210> 68 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 68 Thr Lys Glu Arg Asp Tyr Tyr Tyr Asp Ser Ser Gly Ser Leu Asp Tyr 1 5 10 15 <210> 69 <211> 321 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 69 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagggtgacc 60 gatcacctgc agggccagcc agaccatcaac aactacctga actggtacca gcagaagccc 120 ggcaaggccc ccaagctgct gatctacacc accagcggcc tgcagagcgg cgtgcccagc 180 aggttcagcg gcaggggcag cggcaccgac ttcaccctgg ccatcagcag cctgcagccc 240 gaggacttcg ccacctacta ctgccagcag agctaccaca gccccttcac cttcggcccc 300 ggcaccaagg tggacatcaa g 321<210> 70<211> 107<212> PRT<213> Artificial Sequence<220><223> Synthetic<400> 70Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Thr Ile Asn Asn Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Thr Thr Ser Gly Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 atcacctgca gggccagcca gaccatcaac aactacctga actggtacca gcagaagccc 120 ggcaaggccc ccaagctgct gatctacacc accagcggcc tgcagagcgg cgtgcccagc 180 aggttcagcg gcaggggcag cggcaccgac ttcaccctgg ccatcagcag cctgcagccc 240 gaggacttcg ccacctacta ctgccagcag agctaccaca gccccttcac cttcggcccc 300 ggcaccaagg tggacatcaa g 321 <210> 70 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 70 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Thr Ile Asn Asn Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Thr Thr Ser Gly Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Arg Gly Ser Gly Thr Asp Phe Thr Leu Ala Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr His Ser Pro Phe 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 105 <210> 71 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 71 cagaccatca acaactac 18 <210> 72 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 72 Gln Thr Ile Asn Asn Tyr 1 5 <210> 73 <211> 9 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 73 accaccagc 9 <210> 74 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> Synthesis <400> 74 Thr Thr Ser 1 <210> 75 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> Synthesis <400> 75 cagcagagct accacagccc cttcacc 27 <210> 76 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Synthesis <400> 76 Gln Gln Ser Tyr His Ser Pro Phe Thr 1 5 <210> 77 <211> 1350 <212> DNA <213> Artificial Sequence <220> <223> Synthesis <400> 77 caggtgcagc tggtggagag cggcggcggc gtggtgcagc ccggcaggag cctgaggctg 60 agctgcgagg ccagcggctt caccttcagc gactacgacc tgcactgggt gaggcaggcc 120 cccggcaagg gcctggagtg ggtggccttc atcagcaacg acggcagcaa cgagtactac 180 cccaagagcg tgaagggcag gttcagcatc agcagggaca acagcaagaa caccctgtac 240 ctgcagatga acagcctgag ggccgaggac accgccgtgt accactgcac caaggagagg 300 gactactact acgacagcag cggcagcctg gactactggg gccagggcac cctggtgacc 360 gtgagcagcg ccagcaccaa gggccccagc gtgttccccc tggccccctg cagcaggagc 420 accagcgaga gcaccgccgc cctgggctgc ctggtgaagg actacttccc cgagcccgtg 480 accgtgagct ggaacagcgg cgccctgacc agcggcgtgc acaccttccc cgccgtgctg 540 cagagcagcg gcctgtacag cctgagcagc gtggtgaccg tgcccagcag cagcctgggc 600 accaagacct acacctgcaa cgtggaccac aagcccagca acaccaaggt ggacaagagg 660 gtggagagca agtacggccc cccctgcccc ccctgccccg cccccgagtt cctgggcggc 720 cccagcgtgt tcctgttccc ccccaagccc aaggacaccc tgatgatcag caggaccccc 780 gaggtgacct gcgtggtggt ggacgtgagc caggaggacc ccgaggtgca gttcaactgg 840 tacgtggacg gcgtggaggt gcacaacgcc aagaccaagc ccagggagga gcagttcaac 900 agcacctaca gggtggtgag cgtgctgacc gtgctgcacc aggactggct gaacggcaag 960 gagtacaagt gcaaggtgag caacaagggc ctgcccagca gcatcgagaa gaccatcagc 1020 aaggccaagg gccagcccag ggagccccag gtgtacaccc tgccccccag ccaggaggag 1080 atgaccaaga accaggtgag cctgacctgc ctggtgaagg gcttctaccc cagcgacatc 1140 gccgtggagt gggagagcaa cggccagccc gagaacaact acaagaccac cccccccgtg 1200 ctggacagcg acggcagctt cttcctgtac agcaggctga ccgtggacaa gagcaggtgg 1260 caggagggca acgtgttcag ctgcagcgtg atgcacgagg ccctgcacaa ccactacacc 1320 cagaagagcc tgagcctgag cctgggcaag 1350 <210> 78 <211> 450 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 78 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Asp Leu His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Phe Ile Ser Asn Asp Gly Ser Asn Glu Tyr Tyr Pro Lys Ser Val 50 55 60 Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr His Cys 85 90 95 Thr Lys Glu Arg Asp Tyr Tyr Tyr Asp Ser Ser Gly Ser Leu Asp Tyr 100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser 130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val 195 200 205 Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys 210 215 220 Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 260 265 270 Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 435 440 445 Gly Lys 450 <210> 79 <211> 642 <212> DNA <213> Artificial Sequence <220> <223> Synthetic <400> 79 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagggtgacc 60 atcacctgca gggccagcca gaccatcaac aactacctga actggtacca gcagaagccc 120 ggcaaggccc ccaagctgct gatctacacc accagcggcc tgcagagcgg cgtgcccagc 180 aggttcagcg gcaggggcag cggcaccgac ttcaccctgg ccatcagcag cctgcagccc 240 gaggacttcg ccacctacta ctgccagcag agctaccaca gccccttcac cttcggcccc 300 ggcaccaagg tggacatcaa gaggaccgtg gccgccccca gcgtgttcat cttccccccc 360 agcgacgagc agctgaagag cggcaccgcc agcgtggtgt gcctgctgaa caacttctac 420 cccagggagg ccaaggtgca gtggaaggtg gacaacgccc tgcagagcgg caacagccag 480 gagagcgtga ccgagcagga cagcaaggac agcacctaca gcctgagcag caccctgacc 540 ctgagcaagg ccgactacga gaagcacaag gtgtacgcct gcgaggtgac ccaccagggc 600 ctgagcagcc ccgtgaccaa gagcttcaac aggggcgagt gc 642 <210> 80 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 80 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Thr Ile Asn Asn Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Thr Thr Ser Gly Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Arg Gly Ser Gly Thr Asp Phe Thr Leu Ala Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr His Ser Pro Phe 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 81 <211> 220 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 81 Met Glu Leu Gly Leu Gly Gly Leu Ser Thr Leu Ser His Cys Pro Trp 1 5 10 15 Pro Arg Gln Gln Pro Ala Leu Trp Pro Thr Leu Ala Ala Leu Ala Leu 20 25 30 Leu Ser Ser Val Ala Glu Ala Ser Leu Gly Ser Ala Pro Arg Ser Pro 35 40 45 Ala Pro Arg Glu Gly Pro Pro Pro Val Leu Ala Ser Pro Ala Gly His 50 55 60 Leu Pro Gly Gly Arg Thr Ala Arg Trp Cys Ser Gly Arg Ala Arg Arg 65 70 75 80 Pro Pro Pro Gln Pro Ser Arg Pro Ala Pro Pro Pro Pro Ala Pro Pro 85 90 95 Ser Ala Leu Pro Arg Gly Gly Arg Ala Ala Arg Ala Gly Gly Pro Gly 100 105 110 Ser Arg Ala Arg Ala Ala Gly Ala Arg Gly Cys Arg Leu Arg Ser Gln 115 120 125 Leu Val Pro Val Arg Ala Leu Gly Leu Gly His Arg Ser Asp Glu Leu 130 135 140 Val Arg Phe Arg Phe Cys Ser Gly Ser Cys Arg Arg Ala Arg Ser Pro 145 150 155 160 His Asp Leu Ser Leu Ala Ser Leu Leu Gly Ala Gly Ala Leu Arg Pro 165 170 175 Pro Pro Gly Ser Arg Pro Val Ser Gln Pro Cys Cys Arg Pro Thr Arg 180 185 190 Tyr Glu Ala Val Ser Phe Met Asp Val Asn Ser Thr Trp Arg Thr Val 195 200 205 Asp Arg Leu Ser Ala Thr Ala Cys Gly Cys Leu Gly 210 215 220 <210> 82 <211> 141 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 82 Ala Gly Gly Pro Gly Ser Arg Ala Arg Ala Ala Gly Ala Arg Gly Cys 1 5 10 15 Arg Leu Arg Ser Gln Leu Val Pro Val Arg Ala Leu Gly Leu Gly His 20 25 30 Arg Ser Asp Glu Leu Val Arg Phe Arg Phe Cys Ser Gly Ser Cys Arg 35 40 45 Arg Ala Arg Ser Pro His Asp Leu Ser Leu Ala Ser Leu Leu Gly Ala 50 55 60 Gly Ala Leu Arg Pro Pro Pro Gly Ser Arg Pro Val Ser Gln Pro Cys 65 70 75 80 Cys Arg Pro Thr Arg Tyr Glu Ala Val Ser Phe Met Asp Val Asn Ser 85 90 95 Thr Trp Arg Thr Val Asp Arg Leu Ser Ala Thr Ala Cys Gly Cys Leu 100 105 110 Gly Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Gly Gly Glu Gln Lys 115 120 125 Leu Ile Ser Glu Glu Asp Leu His His His His His His 130 135 140 <210> 83 <211> 141 <212> PRT <213> Artificial Sequence <220> <223> Synthesis <400> 83 Ala Gly Gly Pro Gly Ser Arg Pro Arg Ala Ala Gly Ala Arg Gly Cys 1 5 10 15 Arg Leu Arg Ser Gln Leu Val Pro Val Arg Ala Leu Gly Leu Gly His 20 25 30 Arg Ser Asp Glu Leu Val Arg Phe Arg Phe Cys Ser Gly Ser Cys Arg 35 40 45 Arg Ala Arg Ser Pro His Asp Leu Ser Leu Ala Ser Leu Leu Gly Ala 50 55 60 Gly Ala Leu Arg Pro Pro Pro Gly Ser Arg Pro Ile Ser Gln Pro Cys 65 70 75 80 Cys Arg Pro Thr Arg Tyr Glu Ala Val Ser Phe Met Asp Val Asn Ser 85 90 95 Thr Trp Arg Thr Val Asp Arg Leu Ser Ala Thr Ala Cys Gly Cys Leu 100 105 110 Gly Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Gly Gly Glu Gln Lys 115 120 125 Leu Ile Ser Glu Glu Asp Leu His His His His His His 130 135 140 <210> 84 <211> 141 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 84 Ala Gly Thr Arg Ser Ser Arg Ala Arg Thr Thr Asp Ala Arg Gly Cys 1 5 10 15 Arg Leu Arg Ser Gln Leu Val Pro Val Ser Ala Leu Gly Leu Gly His 20 25 30 Ser Ser Asp Glu Leu Ile Arg Phe Arg Phe Cys Ser Gly Ser Cys Arg 35 40 45 Arg Ala Arg Ser Gln His Asp Leu Ser Leu Ala Ser Leu Leu Gly Ala 50 55 60 Gly Ala Leu Arg Ser Pro Pro Gly Ser Arg Pro Ile Ser Gln Pro Cys 65 70 75 80 Cys Arg Pro Thr Arg Tyr Glu Ala Val Ser Phe Met Asp Val Asn Ser 85 90 95 Thr Trp Arg Thr Val Asp His Leu Ser Ala Thr Ala Cys Gly Cys Leu 100 105 110 Gly Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Gly Gly Glu Gln Lys 115 120 125 Leu Ile Ser Glu Glu Asp Leu His His His His His His 130 135 140 <210> 85 <211> 578 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 85 Asp Pro Leu Pro Thr Glu Ser Arg Leu Met Asn Ser Cys Leu Gln Ala 1 5 10 15 Arg Arg Lys Cys Gln Ala Asp Pro Thr Cys Ser Ala Ala Tyr His His 20 25 30 Leu Asp Ser Cys Thr Ser Ser Ile Ser Thr Pro Leu Pro Ser Glu Glu 35 40 45 Pro Ser Val Pro Ala Asp Cys Leu Glu Ala Ala Gln Gln Leu Arg Asn 50 55 60 Ser Ser Leu Ile Gly Cys Met Cys His Arg Arg Met Lys Asn Gln Val 65 70 75 80 Ala Cys Leu Asp Ile Tyr Trp Thr Val His Arg Ala Arg Ser Leu Gly 85 90 95 Asn Tyr Glu Leu Asp Val Ser Pro Tyr Glu Asp Thr Val Thr Ser Lys 100 105 110 Pro Trp Lys Met Asn Leu Ser Lys Leu Asn Met Leu Lys Pro Asp Ser 115 120 125 Asp Leu Cys Leu Lys Phe Ala Met Leu Cys Thr Leu Asn Asp Lys Cys 130 135 140 Asp Arg Leu Arg Lys Ala Tyr Gly Glu Ala Cys Ser Gly Pro His Cys 145 150 155 160 Gln Arg His Val Cys Leu Arg Gln Leu Leu Thr Phe Phe Glu Lys Ala 165 170 175 Ala Glu Pro His Ala Gln Gly Leu Leu Leu Cys Pro Cys Ala Pro Asn 180 185 190 Asp Arg Gly Cys Gly Glu Arg Arg Arg Asn Thr Ile Ala Pro Asn Cys 195 200 205 Ala Leu Pro Pro Val Ala Pro Asn Cys Leu Glu Leu Arg Arg Leu Cys 210 215 220 Phe Ser Asp Pro Leu Cys Arg Ser Arg Leu Val Asp Phe Gln Thr His 225 230 235 240 Cys His Pro Met Asp Ile Leu Gly Thr Cys Ala Thr Glu Gln Ser Arg 245 250 255 Cys Leu Arg Ala Tyr Leu Gly Leu Ile Gly Thr Ala Met Thr Pro Asn 260 265 270 Phe Val Ser Asn Val Asn Thr Ser Val Ala Leu Ser Cys Thr Cys Arg 275 280 285 Gly Ser Gly Asn Leu Gln Glu Glu Cys Glu Met Leu Glu Gly Phe Phe 290 295 300 Ser His Asn Pro Cys Leu Thr Glu Ala Ile Ala Ala Lys Met Arg Phe 305 310 315 320 His Ser Gln Leu Phe Ser Gln Asp Trp Pro His Pro Thr Phe Ala Val 325 330 335 Met Ala His Gln Asn Glu Asn Pro Ala Val Arg Pro Gln Pro Trp Asp 340 345 350 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 355 360 365 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 370 375 380 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 385 390 395 400 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 405 410 415 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 420 425 430 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 435 440 445 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 450 455 460 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 465 470 475 480 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 485 490 495 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 500 505 510 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 515 520 525 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 530 535 540 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 545 550 555 560 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 565 570 575 Gly Lys

Claims

1. An isolated monoclonal antibody or antigen-binding fragment thereof that specifically binds to human artemin, wherein said antibody or antigen-binding fragment comprises: (a) An HCDR1 consisting of the amino acid sequence of SEQ ID NO:4, an HCDR2 consisting of the amino acid sequence of SEQ ID NO:6, an HCDR3 consisting of the amino acid sequence of SEQ ID NO:8, an LCDR1 consisting of the amino acid sequence of SEQ ID NO:12, an LCDR2 consisting of the amino acid sequence of SEQ ID NO:14, and an LCDR3 consisting of the amino acid sequence of SEQ ID NO:16; or (b) An HCDR1 consisting of the amino acid sequence of SEQ ID NO:24, an HCDR2 consisting of the amino acid sequence of SEQ ID NO:26, an HCDR3 consisting of the amino acid sequence of SEQ ID NO:28, an LCDR1 consisting of the amino acid sequence of SEQ ID NO:32, an LCDR2 consisting of the amino acid sequence of SEQ ID NO:34, and an LCDR3 consisting of the amino acid sequence of SEQ ID NO:36; or (c) An HCDR1 consisting of the amino acid sequence of SEQ ID NO:44, an HCDR2 consisting of the amino acid sequence of SEQ ID NO:46, an HCDR3 consisting of the amino acid sequence of SEQ ID NO:48, an LCDR1 consisting of the amino acid sequence of SEQ ID NO:52, an LCDR2 consisting of the amino acid sequence of SEQ ID NO:54, and an LCDR3 consisting of the amino acid sequence of SEQ ID NO:56; or (d) An HCDR1 consisting of the amino acid sequence of SEQ ID NO:64, an HCDR2 consisting of the amino acid sequence of SEQ ID NO:66, an HCDR3 consisting of the amino acid sequence of SEQ ID NO:68, an LCDR1 consisting of the amino acid sequence of SEQ ID NO:72, an LCDR2 consisting of the amino acid sequence of SEQ ID NO:74, and an LCDR3 consisting of the amino acid sequence of SEQ ID NO:

76.

2. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein said antibody or antigen-binding fragment comprises: An HCDR1 consisting of the amino acid sequence of SEQ ID NO:64, an HCDR2 consisting of the amino acid sequence of SEQ ID NO:66, an HCDR3 consisting of the amino acid sequence of SEQ ID NO:68, an LCDR1 consisting of the amino acid sequence of SEQ ID NO:72, an LCDR2 consisting of the amino acid sequence of SEQ ID NO:74, and an LCDR3 consisting of the amino acid sequence of SEQ ID NO:

76.

3. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 2, wherein the antibody or antigen-binding fragment thereof comprises: an HCVR consisting of the amino acid sequence of SEQ ID NO:62 and an LCVR consisting of the amino acid sequence of SEQ ID NO:

70.

4. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 2, wherein the antibody or antigen-binding fragment thereof comprises: a heavy chain consisting of the amino acid sequence of SEQ ID NO:78 and a light chain consisting of the amino acid sequence of SEQ ID NO:

80.

5. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof comprises: An HCDR1 consisting of the amino acid sequence of SEQ ID NO:4, an HCDR2 consisting of the amino acid sequence of SEQ ID NO:6, an HCDR3 consisting of the amino acid sequence of SEQ ID NO:8, an LCDR1 consisting of the amino acid sequence of SEQ ID NO:12, an LCDR2 consisting of the amino acid sequence of SEQ ID NO:14, and an LCDR3 consisting of the amino acid sequence of SEQ ID NO:

16.

6. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 5, wherein the antibody or antigen-binding fragment thereof comprises: an HCVR consisting of the amino acid sequence of SEQ ID NO:2 and an LCVR consisting of the amino acid sequence of SEQ ID NO:

10.

7. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 5, wherein the antibody or antigen-binding fragment thereof comprises: a heavy chain consisting of the amino acid sequence of SEQ ID NO:18 and a light chain consisting of the amino acid sequence of SEQ ID NO:

20.

8. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof comprises: An HCDR1 consisting of the amino acid sequence of SEQ ID NO:24, an HCDR2 consisting of the amino acid sequence of SEQ ID NO:26, an HCDR3 consisting of the amino acid sequence of SEQ ID NO:28, an LCDR1 consisting of the amino acid sequence of SEQ ID NO:32, an LCDR2 consisting of the amino acid sequence of SEQ ID NO:34, and an LCDR3 consisting of the amino acid sequence of SEQ ID NO:

36.

9. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 8, wherein the antibody or antigen-binding fragment thereof comprises: an HCVR consisting of the amino acid sequence of SEQ ID NO:22 and an LCVR consisting of the amino acid sequence of SEQ ID NO:

30.

10. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 8, wherein the antibody or antigen-binding fragment thereof comprises: a heavy chain consisting of the amino acid sequence of SEQ ID NO:38 and a light chain consisting of the amino acid sequence of SEQ ID NO:

40.

11. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof comprises: An HCDR1 consisting of the amino acid sequence of SEQ ID NO:44, an HCDR2 consisting of the amino acid sequence of SEQ ID NO:46, an HCDR3 consisting of the amino acid sequence of SEQ ID NO:48, an LCDR1 consisting of the amino acid sequence of SEQ ID NO:52, an LCDR2 consisting of the amino acid sequence of SEQ ID NO:54, and an LCDR3 consisting of the amino acid sequence of SEQ ID NO:

56.

12. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 11, wherein the antibody or antigen-binding fragment thereof comprises: an HCVR consisting of the amino acid sequence of SEQ ID NO:42 and an LCVR consisting of the amino acid sequence of SEQ ID NO:

50.

13. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 11, wherein the antibody or antigen-binding fragment thereof comprises: a heavy chain consisting of the amino acid sequence of SEQ ID NO:58 and a light chain consisting of the amino acid sequence of SEQ ID NO:

60.

14. The isolated monoclonal antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody or antigen-binding fragment thereof is fully human.

15. A pharmaceutical composition comprising the monoclonal antibody or antigen-binding fragment thereof according to any one of claims 1 to 14 and a pharmaceutically acceptable carrier.

16. A nucleic acid molecule comprising a nucleotide sequence encoding the monoclonal antibody or antigen-binding fragment thereof as claimed in claim 1.

17. An expression vector comprising the nucleic acid molecule as claimed in claim 16.

18. A host cell comprising the expression vector as claimed in claim 17.

19. A method for producing an anti-human artemin antibody or antigen-binding fragment thereof, the method comprising introducing the expression vector as claimed in claim 17 into a host cell, growing the host cell under conditions permitting production of the antibody or antigen-binding fragment thereof, and recovering the produced antibody or antigen-binding fragment.

20. Use of the monoclonal antibody or antigen-binding fragment thereof as claimed in claim 1 in the preparation of a medicament for reducing thermal hyperalgesia in a subject in need thereof.

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