Materials and methods for treating stress-related disorders and cancer

Abstract

Disclosed herein are materials and methods for the treatment of stress-related disorders and cancer. Methods of inhibiting the hypothalamic pituitary adrenal (HPA) axis in a subject and methods of reducing corticotropin releasing hormone, adrenocorticotropin releasing hormone, and / or cortisol in a subject in need thereof are also provided.

Description

MATERIALS AND METHODS FOR TREATING STRESS-RELATED DISORDERS AND CANCERSTATEMENT OF GOVERNMENT SUPPORT

[0001] This invention was made with government support under Grant No. AG064942, awarded by the National Institutes of Health. The government has certain rights in the inventionFIELD OF THE INVENTION

[0002] The present disclosure is directed to humanized anti-CRH antibodies and the use of such antibodies for the treatment of disorders associated with hypothalamic-pituitary adrenal (HPA) axis activation (e.g. stress- related disorders, obesity, diabetes and Alzheimer's Disease).INCORPORATION BY REFERENCE OF MATERIALS SUBMITTED ELECTRONICALLY

[0003] This application contains, as a separate part of the disclosure, a Sequence Listing in computer readable form ( Filename: 70049_SeqListing.xml; Size: 239,367 bytes; Created: August 29, 2025), which is incorporated by reference in its entirety.BACKGROUND

[0004] Corticotropin-releasing hormone (CRH) is a central coordinator of the neuroendocrine and behavioral response to stressful stimuli. CRH has been evolutionarily conserved in the vertebrate lineage from a time preceding the teleosts and tetrapods, over 550 million years. This conservation across species highlights the importance of CRH in the vertebrate response to stressful stimuli.

[0005] CRH signals through its receptor the CRHR1 to control hypothalamic-pituitary-adrenal (HPA) axis activation; CRH release from the paraventricular nucleus (PVN) of the hypothalamus acts on CRHR1 receptors in the anterior pituitary, causing Adrenocorticotropic Hormone (ACTH) release that stimulates the adrenal glands to produce and release glucocorticoids (GCs), namely cortisol in humans and corticosterone in rodents. Glucocorticoids freely diffuse throughout the body and act on high affinity mineralocorticoid (MR) and low affinity glucocorticoid (GR) receptors that are expressed in almost every vertebrate cell type. MR and GR signaling cause both rapid functional responses in many cell types in addition to long-standing effects through regulation of transcription via glucocorticoid-response elements (GREs). The response to GR and MR receptor activation leads to mobilization of resources and cellular adaptations that prepare the body to overcome a stressor.

[0006] In the absence of a stressful stimulus, the PVN receives input from the suprachiasmatic nucleus (SON) of the hypothalamus and responds to melatonin levels, altering CRH release throughout the day based on these inputs. These daily fluctuations in CRH release in turn produce the circadian rhythm of GC release, with peak levels of GCs being present at awakening and trough levels before returning to sleep.

[0007] In addition to its effects on the HPA Axis, CRH and the CRHR1 are widely expressed in many regions of the brain implicated in cognition and anxiety, including the neocortex, hippocampus, amygdala, and locus coeruleus. CRH has been shown to have region specific effects on anxiety, and is best described as aneuroregulator: augmenting the activity of CRHR1 expressing neurons but not serving as a classical neurotransmitter.SUMMARY

[0008] In one aspect, the disclosure provides an isolated antibody or antigen binding fragment thereof that binds to corticotropin releasing hormone (CRH) comprising three heavy chain complementarity determining regions (CDRs) (CDR-H1, CDR-H2, and CDR-H3) and three light chain complementarity determining regions (CDRs) (CDR-L1, CDR-L2, and CDR-L3), wherein the CDR-H1, CDR-H2, and CDR-H3 are present in a heavy chain variable domain (VH) comprising the amino acid sequence set forth in any one of SEQ ID NOs:163-193 and wherein the CDR-L1, CDR-L2, and CDR-L3 are present in a light chain variable domain (VL) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 194-238. In some embodiments, the CDR-H1, CDR- H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to Kabat. In some embodiments, the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to Chothia. In some embodiments, the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to IMGT. In some embodiments, the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to AbM. In some embodiments, the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to Contact. In some embodiments, the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to Honneger (AHo).

[0009] In another aspect, the disclosure provides antibody or antigen-binding fragment thereof comprising a set of 6 CDRs, wherein the CDRs comprise: (a) a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 1 (DSTMX), SEQ ID NO: 2 (DSTIX) or DSTXX; (b) a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 10 (XIHPDXGGTXYXQKFXG) or SEQ ID NO: 239 (XIHPDSGGTIYNQKFQG); (c) a CDR- H3 comprising the amino acid sequence set forth in SEQ ID NO: 29 (XGFAY), SEQ ID NO: 30 (XPFAY), SEQ ID NO: 31 (NGXAY), SEQ ID NO: 32 (NGFXY), or SEQ ID NO: 33 (TXFAY); (d) a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 46 (RSSXSLLGSXXXTXLX); (e) a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 62 (XVSNKXX); and (f) a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 70 (SQXXXHXPX).

[0010] Methods of treating a disorder associated with HPA axis activation (e.g., a stress-related disorder or cancer) are contemplated. For example, in some embodiments, described herein is a method of treating a stress-related disorder. In this regard, the method comprises administering to a subject in need thereof an antibody or antigen binding fragment thereof described herein in an amount effective to treat the disorder. Exemplary stress-related disorders include, but are not limited to, obesity, diabetes, anxiety, depression, Alzheimer's Disease, post traumatic stress disorder, generalized anxiety disorder, major depression, anorexia nervosa, adrenal disorder, metabolic syndrome, type 1 diabetes, sarcopenia and multiple sclerosis.DETAILED DESCRIPTION

[0011] In one aspect, the disclosure provides an antibody (or antigen binding fragment thereof) that binds corticotropin releasing hormone (CRH) comprises three heavy chain complementarity determining regions (CDRs) (CDR-H1, CDR-H2, and CDR-H3) and three light chain complementarity determining regions (CDRs) (CDR-L1, CDR-L2, and CDR-L3), wherein the CDR-H1, CDR-H2, and CDR-H3 are present in a heavy chain variable domain (VH) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 164-193 and wherein the CDR-L1, CDR-L2, and CDR-L3 are present in a light chain variable domain (VL) comprising the amino acid sequence set forth in any one of SEQ ID NO: 194-238.

[0012] Corticotropin-releasing hormone (CRH) is a 41 -amino acid peptide (SEEPPISLDLTFHLLREVLEMARAEQLAQQAHSNRKLMEII (SEQ ID NO: 88)) derived from a 196-amino acid preprohormone (Genbank Accession No. EAW86897.1). CRH is secreted by the paraventricular nucleus (PVN) of the hypothalamus in response to stress. Increased CRH production has been observed to be associated with Alzheimer's disease and major depression (Raadsheer et al., Am J Psychiatry.152 (9): 1372-6, 1995), and autosomal recessive hypothalamic corticotropin deficiency has multiple (and potentially fatal) metabolic consequences, including hypoglycemia.

[0013] The term "antibody” refers to an intact immunoglobulin molecule (including polyclonal, monoclonal, chimeric, humanized, and / or human versions having full length heavy and / or light chains). The antibody may be any type of antibody, i.e., immunoglobulin, known in the art. In exemplary embodiments, the antibody is an antibody of class or isotype IgA, IgD, IgE, IgG, or IgM. In exemplary embodiments, the antibody described herein comprises one or more alpha, delta, epsilon, gamma, and / or mu heavy chains. In exemplary embodiments, the antibody described herein comprises one or more kappa or light chains. In exemplary aspects, the antibody is an IgG antibody and optionally is one of the four human subclasses: lgG1, lgG2, lgG3 and lgG4. In some embodiments, the antibody is an lgG1.

[0014] The term "humanized" refers to antibodies having at least CDR regions from a non-human source and which are engineered to have a structure and immunological function more similar to true human antibodies than the original source antibodies. For example, humanizing can involve grafting CDRs from a non-human antibody, such as a mouse antibody, into a human antibody framework. Humanizing also can involve select amino acid substitutions to make a non-human sequence look more like a human sequence.

[0015] "Specifically binds” as used herein means that the antibody (or antigen binding fragment) preferentially binds an antigen (CRH peptide) over other proteins. In some embodiments, "specifically binds” means the antibody has a higher affinity for the antigen than for other proteins. Antibodies that specifically bind an antigen may have a binding affinity for the antigen of less than or equal to 1 x 107M, less than or equal to 2 x 107M, less than or equal to 3 x 107M, less than or equal to 4 x 107M, less than or equal to 5 x 107M, less than or equal to 6 x 107M, less than or equal to 7 x 107M, less than or equal to 8 x 107M, less than or equal to 9 x 107M, less than or equal to 1 x 108M, less than or equal to 2 x KT8M, less than or equal to 3 x 10-8M, lessthan or equal to 4 x 10’8M, less than or equal to 5 x 10’8M, less than or equal to 6 x 10’8M, less than or equal to 7 x 10’8M, less than or equal to 8 x 10’8M, less than or equal to 9 x 10’8M, less than or equal to 1 x 109M, less than or equal to 2 x 10’9M, less than or equal to 3 x 10’9M, less than or equal to 4 x 10’9M, less than or equal to 5 x 10'9M, less than or equal to 6 x 10'9M, less than or equal to 7 x 10'9M, less than or equal to 8 x 10'9M, less than or equal to 9 x 10'9M, less than or equal to 1 x 10'10M, less than or equal to 2 x 10'10M, less than or equal to 3 x 10'10M, less than or equal to 4 x 10'10M, less than or equal to 5 x 10'10M, less than or equal to 6 x 10'10M, less than or equal to 7 x 10'10M, less than or equal to 8 x 10'10M, less than or equal to 9 x 10'10M, less than or equal to 1 x 10’11M, less than or equal to 2 x 10’11M, less than or equal to 3 x 10’11M, less than or equal to 4 x 10’11M, less than or equal to 5 x 10’11M, less than or equal to 6 x 10’11M, less than or equal to 7 x 10’11M, less than or equal to 8 x 10’11M, less than or equal to 9 x 10’11M, less than or equal to 1 x 10’12M, less than or equal to 2 x 10’12M, less than or equal to 3 x 10’12M, less than or equal to 4 x 10’12M, less than or equal to 5 x 10’12M, less than or equal to 6 x 10’12M, less than or equal to 7 x 10’12M, less than or equal to 8 x 10’12M, or less than or equal to 9 x 1012M. It will be appreciated that ranges having the values above as end points is contemplated in the context of the disclosure. For example, the antibody or antigen binding fragment thereof may bind CRH of SEQ ID NO: XX with an affinity of about 1 x 107M to about 9 x 10 '12M or an affinity of 1 x 109to about 9 x 1012.

[0016] In some or any embodiments, the antibody (or antigen binding fragment) binds to CRH of SEQ ID NO: 88, or a naturally occurring variant thereof, with an affinity (Kd) of less than or equal to 1 x 107M, less than or equal to 1 x 108M, less than or equal to 1 x 109M, less than or equal to 1 x 10’10M, less than or equal to 1 x 10 -11M, or less than or equal to 1 x 1012M, or ranging from 1 x 109to 1 xIO10, or ranging from 1 x 1012to about 1 x 1013. Affinity is determined using a variety of techniques, examples of which include an affinity ELISA assay and a surface plasmon resonance (BIAcore) assay.

[0017] "CDR" refers to the complementarity determining region within antibody variable sequences. There are three CDRs in each of the variable regions of the heavy chain and the light chain, which are designated CDR1 , CDR2 and CDR3, for each of the variable regions. The term "set of six CDRs " as used herein refers to a group of three CDRs that occur in the light chain variable region and heavy chain variable region, which are capable of binding the antigen. The exact boundaries of CDRs have been defined differently according to different systems. The system described by Kabat (Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987) and (1991)) not only provides an unambiguous residue numbering system applicable to any variable region of an antibody, but also provides precise residue boundaries defining the three CDRs. These CDRs may be referred to as Kabat CDRs. Chothia and coworkers (Chothia & Lesk, J. Mol. Biol. 196:901-917 (1987) and Chothia et al., Nature 342:877-883 (1989)) found that certain sub-portions within Kabat CDRs adopt nearly identical peptide backbone conformations, despite having great diversity at the level of amino acid sequence. These sub-portions were designated as L1 , L2 and L3 or H1 , H2 and H3 where the "L" and the "H" designates the light chain and the heavy chains regions, respectively. These regions may be referred to as Chothia CDRs, which have boundaries that overlap with Kabat CDRs. Other boundaries defining CDRs overlapping with the Kabat CDRs have been described by Padlan (FASEB J.9:133-139 (1995)) and MacCallum (J Mol Biol 262(5)73245 (1996)). Still other CDR boundary definitions may not strictly follow one of the above systems, but will nonetheless overlap with the Kabat CDRs, although they may be shortened or lengthened in light of prediction or experimental findings that particular residues or groups of residues or even entire CDRs do not significantly impact antigen binding. The methods used herein may utilize CDRs defined according to any of these systems, although preferred embodiments use Kabat or Chothia defined CDRs.

[0018] CDRs are obtained by, e.g., constructing polynucleotides that encode the CDR of interest and expression in a suitable host cell. Such polynucleotides are prepared, for example, by using the polymerase chain reaction to synthesize the variable region using mRNA of antibody-producing cells as a template (see, for example, Larrick et al., Methods: A Companion to Methods in Enzymology, 2:106 (1991); Courtenay-Luck, "Genetic Manipulation of Monoclonal Antibodies," in Monoclonal Antibodies Production, Engineering and Clinical Application, Ritter et al. (eds.), page 166, Cambridge University Press (1995); and Ward et al., "Genetic Manipulation and Expression of Antibodies," in Monoclonal Antibodies: Principles and Applications, Birch et al., (eds.), page 137, Wiley-Liss, Inc. (1995)).

[0019] The exact definitional CDR boundaries and lengths are subject to different classification and numbering systems. CDRs may therefore be referred to by Kabat, Chothia, contact or any other boundary definitions, including the numbering system described herein. Despite differing boundaries, each of these systems has some degree of overlap in what constitutes the so-called "hypervariable regions” within the variable sequences. CDR definitions according to these systems may therefore differ in length and boundary areas with respect to the adjacent framework region. See for example Kabat (an approach based on cross-species sequence variability), Chothia (an approach based on crystallographic studies of antigen-antibody complexes), MacCallum, Honegger, and IMGT (Wu and Kabat, J. Exp. Med. 1970; 132:211-250; Chothia et al., J. Mol. Biol, 1987, 196: 901-917; MacCallum et al., J. Mol. Biol, 1996, 262: 732; Lefranc et al., Dev. Comp. Immunol. 2003; 27:55-77; Honegger et al., J. Mol. Bol. (2001); 309:657-670). Still another standard for characterizing the antigen binding side is the AbM definition used by Oxford Molecular's AbM antibody modeling software. See, e.g., Protein Sequence and Structure Analysis of Antibody Variable Domains. In: Antibody Engineering Lab Manual (Ed.: Duebel, S. and Kontermann, R., Springer-Verlag, Heidelberg). To the extent that two residue identification techniques define regions of overlapping, but not identical regions, they can be combined to define a hybrid CDR. In some embodiments, the CDRs disclosed herein are identified according to the Kabat numbering system. In some embodiments, the CDRs disclosed herein are identified according to the Chothia numbering system. In some embodiments, the CDRs disclosed herein are identified according to the IMGT approach. In some embodiments, the CDRs disclosed herein are identified according to the Honegger approach.

[0020] In some embodiments, the CDRs of the antibodies described herein comprise(a) a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 1 (DSTMX), SEQ ID NO: 2 (DSTIX) or DSTXX;(b) a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 10 (XIHPDXGGTXYXQKFXG)or SEQ ID NO: 239 (XIHPDSGGTIYNQKFQG);(c) a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 29 (XGFAY), SEQ ID NO: 30 (XPFAY), SEQ ID NO: 31 (NGXAY), SEQ ID NO: 32 (NGFXY), or SEQ ID NO: 33 (TXFAY);(d) a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 46 (RSSXSLLGSXXXTXLX);(e) a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 62 (XVSNKXX); and(f) a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 70 (SQXXXHXPX).

[0021] In some embodiments, the antibody (or antigen binding fragment thereof) comprises a CDR-H1 comprising the amino acid sequence set forth in any one of SEQ ID NOs: 3-9. In some embodiments, the antibody (or antigen binding fragment thereof) comprises a CDR-H2 comprising the amino acid sequence set forth in any one of SEQ ID NOs: 11-28 or 239. In some embodiments, the antibody (or antigen binding fragment thereof) comprises a CDR-H3 comprising the amino acid sequence set forth in any one of SEQ ID NOs: 34-45. In some embodiments, the antibody (or antigen binding fragment thereof) comprises a CDR-L1 comprising the amino acid sequence set forth in any one of SEQ ID NOs: 47-61 . In some embodiments, the antibody (or antigen binding fragment thereof) comprises a CDR-L2 comprising the amino acid sequence set forth in any one of SEQ ID NOs: 63-69. In some embodiments, the antibody (or antigen binding fragment thereof) comprises a CDR-L3 comprising the amino acid sequence set forth in any one of SEQ ID NOs: 71-85.

[0022] In some embodiments, the antibody (or antigen binding fragment thereof) comprises a set of six CDRs as follows: CDR-H1 of SEQ ID NO: 4, CDR-H2 of SEQ ID NO: 12, CDR-H3 of SEQ ID NO: 34, CDR-L1 of SEQ ID NO: 47, CDR-L2 of SEQ ID NO: 63 and CDR-L3 of SEQ ID NO: 71 .

[0023] In some embodiments, the antibody (or antigen binding fragment thereof) comprises a set of six CDRs as follows: CDR-H1 of SEQ ID NO: 5, CDR-H2 of SEQ ID NO: 13, CDR-H3 of SEQ ID NO: 34, CDR-L1 of SEQ ID NO: 47, CDR-L2 of SEQ ID NO: 63 and CDR-L3 of SEQ ID NO: 71 .

[0024] In some embodiments, the antibody (or antigen binding fragment thereof) comprises a set of six CDRs as follows: CDR-H1 of SEQ ID NO: 5, CDR-H2 of SEQ ID NO: 14, CDR-H3 of SEQ ID NO: 34, CDR-L1 of SEQ ID NO: 47, CDR-L2 of SEQ ID NO: 63 and CDR-L3 of SEQ ID NO: 71 .

[0025] In some embodiments, the antibody (or antigen binding fragment thereof) comprises a set of six CDRs as follows: CDR-H1 of SEQ ID NO: 5, CDR-H2 of SEQ ID NO: 239, CDR-H3 of SEQ ID NO: 34, CDR-L1 of SEQ ID NO: 47, CDR-L2 of SEQ ID NO: 63 and CDR-L3 of SEQ ID NO: 71 .

[0026] In some embodiments, the antibody (or antigen binding fragment thereof) comprises a heavy chain variable region comprising an amino acid sequence having at least 75% identity (e.g., at least 75%, 80%, 85%, 90%, 95% or 100% identity) to an amino acid sequence set forth in any one of SEQ ID NO: 163-193. In some embodiments, the antibody (or antigen binding fragment thereof) comprises a heavy chain variable region comprising an amino acid sequence set forth in any one of SEQ ID NO: 163-193. In some embodiments, the antibody (or antigen binding fragment thereof) comprises a light chain variable region comprising an amino acid sequence having at least 75% identity (e.g., at least 75%, 80%, 85%, 90%, 95% or 100% identity) to an aminoacid sequence set forth in any one of SEQ ID NO: 194-238. In some embodiments, the antibody (or antigen binding fragment thereof) comprises a light chain variable region comprising an amino acid sequence set forth in any one of SEQ ID NO: 194-238.

[0027] In some embodiments, the antibody (or antigen binding fragment thereof) comprises a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 164 and a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 234 or SEQ ID NO: 236.

[0028] In some embodiments, the antibody (or antigen binding fragment thereof) comprises a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 175 and a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 234, or SEQ ID NO: 236.

[0029] In some embodiments, the antibody (or antigen binding fragment thereof) comprises a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 186 and a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 233, SEQ ID NO: 234, SEQ ID NO: 235, or SEQ ID NO: 236.

[0030] In some embodiments, the antibody (or antigen binding fragment thereof) comprises a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 193 and a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 234 or SEQ ID NO: 236.

[0031] In some embodiments, the antibody (or antigen binding fragment thereof) comprises a heavy chain amino acid sequence having at least 75% identity (e.g., at least 75%, 80%, 85%, 90%, 95% or 100% identity) to an amino acid sequence set forth in any one of SEQ ID NO: 86-116. In some embodiments, the antibody (or antigen binding fragment thereof) comprises a heavy chain amino acid sequence set forth in any one of SEQ ID NOs: 86-116. In some embodiments, the antibody (or antigen binding fragment thereof) comprises a light chain amino acid sequence having at least 75% identity (e.g., at least 75%, 80%, 85%, 90%, 95% or 100% identity) to an amino acid sequence set forth in any one of SEQ ID NO: 117-162. In some embodiments, the antibody (or antigen binding fragment thereof) comprises a heavy chain amino acid sequence set forth in any one of SEQ ID NOs: 117-162.

[0032] In some embodiments, the antibody comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 89 and a light chain amino acid sequence set forth in SEQ ID NO: 119. In some embodiments, the antibody comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 89 and a light chain amino acid sequence set forth in SEQ ID NO: 120. In some embodiments, the antibody comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 89 and a light chain amino acid sequence set forth in SEQ ID NO: 121.

[0033] In some embodiments, the antibody comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 90 and a light chain amino acid sequence set forth in SEQ ID NO: 119. In some embodiments, theantibody comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 90 and a light chain amino acid sequence set forth in SEQ ID NO: 120. In some embodiments, the antibody comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 90 and a light chain amino acid sequence set forth in SEQ ID NO: 121.

[0034] In some embodiments, the antibody comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 91 and a light chain amino acid sequence set forth in SEQ ID NO: 119. In some embodiments, the antibody comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 91 and a light chain amino acid sequence set forth in SEQ ID NO: 120. In some embodiments, the antibody comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 91 and a light chain amino acid sequence set forth in SEQ ID NO: 121.

[0035] In some embodiments, the antibody (or antigen binding fragment thereof) comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 100 and a light chain amino acid sequence set forth in SEQ ID NO: 161. In some embodiments, the antibody (or antigen binding fragment thereof) comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 100 and a light chain amino acid sequence set forth in SEQ ID NO: 158. In some embodiments, the antibody (or antigen binding fragment thereof) comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 118 and a light chain amino acid sequence set forth in SEQ ID NO: 157.

[0036] Antigen Binding Fragments

[0037] Antigen binding fragments of the antibodies described herein are also contemplated. Examples of antigen binding fragments, include, but are not limited to, Fab, F(ab')2, a monospecific or bispecific Fab2, a trispecific Fab3, scFv, dsFv, scFv-Fc, bispecific diabodies, trispecific triabodies, minibodies, a fragment of IgNAR (e.g., V-NAR), a fragment of hcigG (e.g., VhH), bis-scFvs, fragments expressed by a Fab expression library, and the like. In exemplary aspects, the antigen binding fragment is a domain antibody, VhH domain, V-NAR domain, VH domain, VL domain, or the like. Antibody fragments of the disclosure, however, are not limited to these exemplary types of antibody fragments. In exemplary aspects, antigen binding fragment is a Fab fragment. In exemplary aspects, the antigen binding fragment comprises two Fab fragments. In exemplary aspects, the antigen binding fragment comprises two Fab fragments connected via a linker. In exemplary aspects, the antigen binding fragment comprises or is a minibody comprising two Fab fragments. In exemplary aspects, the antigen binding fragment comprises, or is, a minibody comprising two Fab fragments joined via a linker.Minibodies are known in the art. See, e.g., Hu et al., Cancer Res 56: 3055-3061 (1996). In exemplary aspects, the antigen binding fragment comprises or is a minibody comprising two Fab fragments joined via a linker, optionally, comprising an alkaline phosphatase domain.

[0038] A domain antibody comprises a functional binding unit of an antibody, and can correspond to the variable regions of either the heavy (VH) or light (VL) chains of antibodies. A domain antibody can have a molecular weight of approximately 13 kDa, or approximately one-tenth of a full antibody. Domain antibodies may be derived from full antibodies such as those described herein.

[0039] In some embodiments, the antigen binding fragment is a nanobody. Nanobodies, also known as single-domain antibodies, are antibody fragments that can bind to a specific antigen, similar to a whole antibody.Nanobodies are made up of a single monomeric variable antibody domain or single-domain antibody (sdAb).Nanobodies have several advantages over conventional antibodies, including their small size, high stability and solubility, ease of engineering and manufacturing, low immunogenicity in humans, and fast tissue penetration.

[0040] Nanobodies lack light chains, as well as one constant domain of the heavy chain (CH1 domain) present in conventional antibodies. They are commonly called heavy chain antibodies (HCAbs). The antigen specificity of HCAbs is confined to the variable domain of the heavy chain (VHH), and this domain is often referred to as a single-domain antibody (sdAb). In HCAbs, the VHH is considered as the structural and functional equivalent of the Fv fragment of conventional antibodies, with the main difference that it consists only of one variable fragment composed of a single polypeptide chain. As such, it has only approximately half the size (about 15 kDa) of the Fv fragment.

[0041] Methods of Antibody or Antigen Binding Fragment Production

[0042] Suitable methods of making antibodies are known in the art. For instance, standard hybridoma methods are described in, e.g., Harlow and Lane (eds.), Antibodies: A Laboratory Manual, CSH Press (1988), and CA. Janeway et al. (eds.), Immunobiology, 5th Ed., Garland Publishing, New York, NY (2001)). Monoclonal antibodies for use in the methods of the disclosure may be prepared using any technique which provides for the production of antibody molecules by continuous cell lines in culture. These include but are not limited to the hybridoma technique originally described by Koehler and Milstein (Nature 256: 495-497, 1975), the human B-cell hybridoma technique (Kosbor et al., Immunol Today 4:72, 1983; Cote et al., Proc Natl Acad Sci 80: 2026-2030, 1983) and the EBV-hybridoma technique (Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R Liss Inc, New York N.Y., pp 77-96, (1985). Alternatively, other methods, such as EBV-hybridoma methods (Haskard and Archer, J. Immunol. Methods, 74(2), 361-67 (1984), and Roder et al., Methods Enzymol., 121, 140-67 (1986)), and bacteriophage vector expression systems (see, e.g., Huse et al., Science, 246, 1275-81 (1989)) are known in the art. Further, methods of producing antibodies in non-human animals are described in, e.g., U.S. Patents 5,545,806, 5,569,825, and 5,714,352, and U.S. Patent Application Publication No. 2002 / 0197266 Al). Antibodies may also be produced by inducing in vivo production in the lymphocyte population or by screening recombinant immunoglobulin libraries or panels of highly specific binding reagents as disclosed in Orland! et al (Proc Natl Acad Sci 86: 3833-3837; 1989), and Winter G and Milstein C (Nature 349: 293-299, 1991). If the full sequence of the antibody or antigen-binding fragment is known, then methods of producing recombinant proteins may be employed. See, e.g., "Protein production and purification” Nat Methods 5(2): 135-146 (2008). In some embodiments, the antibodies (or antigen binding fragments) are isolated from cell culture or a biological sample if generated in vivo.

[0043] Phage display also can be used to generate the antibodies described herein. In this regard, phage libraries encoding antigen-binding variable (V) domains of antibodies can be generated using standard molecular biology and recombinant DNA techniques (see, e.g., Sambrook et al. (eds.), Molecular Cloning, A Laboratory Manual, 3rd Edition, Cold Spring Harbor Laboratory Press, New York (2001)). Phage encoding a variable regionwith the desired specificity are selected for specific binding to the desired antigen, and a complete or partial antibody is reconstituted comprising the selected variable domain. Nucleic acid sequences encoding the reconstituted antibody are introduced into a suitable cell line, such as a myeloma cell used for hybridoma production, such that antibodies having the characteristics of monoclonal antibodies are secreted by the cell (see, e.g., Janeway et al., supra, Huse et al., supra, and U.S. Patent 6,265,150). Related methods also are described in U.S. Patent No. 5,403,484; U.S. Patent No. 5,571,698; U.S. Patent No. 5,837,500; U.S. Patent No. 5,702,892. The techniques described in U.S. Patent No. 5,780,279; U.S. Patent No. 5,821,047; U.S. Patent No. 5,824,520; U.S. Patent No. 5,855,885; U.S. Patent No. 5,858,657; U.S. Patent No. 5,871,907; U.S. Patent No. 5,969,108; U.S. Patent No. 6,057,098; and U.S. Patent No. 6,225,447.

[0044] Antibodies can be produced by transgenic mice that are transgenic for specific heavy and light chain immunoglobulin genes. Such methods are known in the art and described in, for example U.S. Patent Nos. 5,545,806 and 5,569,825, and Janeway et al., supra.

[0045] Chemically constructed bispecific antibodies may be prepared by chemically cross-linking heterologous Fab or F(ab')2 fragments by means of chemicals such as heterobifunctional reagent succinimidy I-3- (2-pyridyldithiol)-propionate (SPDP, Pierce Chemicals, Rockford, III.). The Fab and F(ab')2 fragments can be obtained from intact antibody by digesting it with papain or pepsin, respectively (Karpovsky et al., J. Exp. Med. 160:1686-701 (1984); Titus et ai., J. Immunol., 138:4018-22 (1987)).

[0046] Methods of testing antibodies for the ability to bind to an epitope of CRH, regardless of how the antibodies are produced, are known in the art and include, e.g., radioimmunoassay (RIA), ELISA, Western blot, immunoprecipitation, surface plasmon resonance (e.g., Biacore), and competitive inhibition assays (see, e.g., Janeway et al., infra, and U.S. Patent Application Publication No. 2002 / 0197266).

[0047] Antibody fragments that contain the antigen binding, or idiotype, of the antibody molecule may be generated by techniques known in the art. For example, a F(ab')2 fragment may be produced by pepsin digestion of the antibody molecule; Fab' fragments may be generated by reducing the disulfide bridges of the F(ab')2 fragment; and two Fab' fragments which may be generated by treating the antibody molecule with papain and a reducing agent. The disclosure is not limited to enzymatic methods of generating antigen binding fragments; the antigen binding fragment may be a recombinant antigen binding fragment produced by expressing a polynucleotide encoding the fragment in a suitable host cell.

[0048] A single-chain variable region fragment (scFv), which consists of a truncated Fab fragment comprising the variable (V) domain of an antibody heavy chain linked to a V domain of an antibody light chain via a synthetic peptide, can be generated using routine recombinant DNA technology techniques (see, e.g., Janeway et al., supra). Similarly, disulfide-stabilized variable region fragments (dsFv) can be prepared by recombinant DNA technology (see, e.g., Reiter et al., Protein Engineering, 7, 697-704 (1994)).

[0049] Recombinant antibody fragments, e.g., scFvs, can also be engineered to assemble into stable multimeric oligomers of high binding avidity and specificity to different target antigens. Such diabodies (dimers),triabodies (trimers) or tetrabodies (tetramers) are well known in the art, see e.g., Kortt et al., Biomol Eng. 2001 18:95-108, (2001) and Todorovska et al., J Immunol Methods. 248:47-66, (2001).

[0050] The heavy chains of the antibodies described herein may further comprise one or more mutations that affect binding of the antibody containing the heavy chains to one or more Fc receptors. One of the functions of the Fc portion of an antibody is to communicate to the immune system when the antibody binds its target. This is commonly referenced as "effector function.” Communication leads to antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and / or complement dependent cytotoxicity (CDC). ADCC and ADCP are mediated through the binding of the Fc to Fc receptors on the surface of cells of the immune system. CDC is mediated through the binding of the Fc with proteins of the complement system, e.g., C1q.

[0051] The effector function of an antibody can be increased, or decreased, by introducing one or more mutations into the Fc. In some embodiments, Fc molecules having increased effector function include those having a mutation (e.g., substitution) at one or more of the following residues [numbering based on the EU numbering scheme]: 228, 234, 235, 236, 237, 238, 239, 248, 249, 252, 254, 255, 256, 258, 265, 267, 268, 269, 270, 272, 279, 280, 282, 283, 285, 298, 289, 290, 292, 293, 294, 295, 296, 297, 298, 301, 303, 305, 307, 312,315, 318, 320, 322, 324, 326, 327, 329, 330, 331, 332, 333, 334, 335, 337, 338, 339, 340, 345, 356, 359, 360,361, 373, 376, 379, 382, 383, 388, 389, 398, 400, 409, 414, 416, 419, 422, 430, 434, 435, 437, 438, 439, 440, or442. In some embodiments, the antibodies described herein comprise an N297A mutation in the Fc region.

[0052] Another method of increasing effector function of IgG Fc-containing proteins is by reducing the fucosylation of the Fc. Removal of the core fucose from the biantennary complex-type oligosachharides attached to the Fc greatly increased ADCC effector function without altering antigen binding or CDC effector function. Several methods are known for reducing or abolishing fucosylation of Fc-containing molecules, e.g., antibodies. These include recombinant expression in certain mammalian cell lines including a FUT8 knockout cell line, variant CHO line Led 3, rat hybridoma cell line YB2 / 0, a cell line comprising a small interfering RNA specifically against the FUT8 gene, and a cell line coexpressing |3-1,4-N-acetylglucosaminyltransferase III and Golgi |3- mannosidase II. Alternatively, the Fc-containing molecule may be expressed in a non-mammalian cell such as a plant cell, yeast, or prokaryotic cell, e.g., E. coll. Thus, in certain embodiments, a composition comprises an antibody having reduced fucosylation or lacking fucosylation altogether.

[0053] In some embodiments, an Fc region or domain can exhibit reduced binding affinity to one or more Fc receptors. In some embodiments, an Fc region or domain can exhibit reduced binding affinity to one or more Fey receptors. In some embodiments, an Fc region or domain can exhibit reduced binding affinity to FcRn receptors. In some embodiments, an Fc region or domain can exhibit reduced binding affinity to Fey and FcRn receptors. In some embodiments, an Fc domain is an Fc null domain or region. As used herein, an "Fc null” refers to a domain that exhibits weak to no binding to any of the Fcgamma receptors. In some embodiments, an Fc null domain or region exhibits a reduction in binding affinity (e.g., increase in Kd) to Fey receptors of at least about 1000-fold.

[0054] The Fc region or domain may have one or more, two or more, three or more, or four or more, or up to five amino acid substitutions that decrease binding of the Fc region or domain to an Fc receptor. In some embodiments, an Fc region or domain exhibits decreased binding to FcyRI (CD64), FcyRIIA (CD32), FcyRIIIA (CD16a), FcyRII I B (CD16b), or any combination thereof. In order to decrease binding affinity of an Fc region or domain to an Fc receptor, an Fc region or domain may comprise one or more amino acid substitutions that has the effect of reducing the affinity of the Fc domain or region to an Fc receptor. In certain embodiments, the Fc region or domain is an lgG1 and the one or more substitutions in the Fc region or domain comprise any one or more of lgG1 heavy chain mutations corresponding to E233P, L234V, L234A, L235A, L235E, AG236, G237A, E318A, K320A, K322A, A327G, A330S, or P331S according to the EU index of Kabat numbering.Detection Methods

[0055] It is sometimes desirable to detect the presence or measure the amount of CRH in a sample. In this regard, the disclosure provides a method of using the antibody or fragment thereof described herein to measure the amount of CRH in a sample. To determine a measurement of CRH, a biological sample from a mammalian subject is contacted with an anti-CRH antibody (or antigen binding fragment thereof) described herein for a time sufficient to allow immunocomplexes to form. Immunocomplexes formed between the antibody and CRH in the sample are then detected. The amount of CRH in the biological sample is optionally quantitated by measuring the amount of the immunocomplex formed between the antibody and the CRH. For example, the antibody can be quantitatively measured if it has a detectable label, or a secondary antibody can be used to quantify the immunocomplex.

[0056] In some embodiments, the biological sample comprises a tissue sample, a cell sample, or a biological fluid sample, such as blood, saliva, serum, or plasma.

[0057] Conditions for incubating an antibody with a test sample vary. Incubation conditions depend on the format employed in the assay, the detection methods employed, and the type and nature of the antibody used in the assay. One skilled in the art will recognize that any one of the commonly available immunological assay formats can readily be adapted to employ the antibodies (or fragments thereof) of the present disclosure. Examples of such assays can be found in Chard, T., An Introduction to Radioimmunoassay and Related Techniques, Elsevier Science Publishers, Amsterdam, The Netherlands (1986); Bullock, G.R. et al., Techniques in Immunocytochemistry, Academic Press, Orlando, FL Vol. 1 (1982), Vol. 2 (1983), Vol. 3 (1985); Tijssen, P., Practice and Theory of immunoassays: Laboratory Techniques in Biochemistry and Molecular Biology, Elsevier Science Publishers, Amsterdam, The Netherlands (1985). The test sample used in the above-described method will vary based on the assay format, nature of the detection method and the tissues, cells or fluids used as the sample to be assayed.

[0058] The assay described herein may be useful in, e.g., evaluating the efficacy of a particular therapeutic treatment regime in animal studies, in clinical trials, or in monitoring the treatment of an individual patient.

[0059] In some embodiments, the anti-CRH antibody (or antigen binding fragment thereof) is attached to a solid support, and binding is detected by detecting a complex between the CRH and the antibody (or antigen binding fragment thereof) on the solid support. The antibody (or antigen binding fragment thereof) optionally comprises a detectable label and binding is detected by detecting the label in the CRH-antibody complex.

[0060] Detection of the presence or absence of a CRH-antibody complex be achieved using any method known in the art. For example, the transcript resulting from a reporter gene transcription assay of a CRH peptide interacting with a target molecule (e.g., antibody) typically encodes a directly or indirectly detectable product (e.g., p-galactosidase activity and luciferase activity). For cell free binding assays, one of the components usually includes, or is coupled to, a detectable label. A wide variety of labels can be used, such as those that provide direct detection (such as radioactivity, luminescence, optical or electron density) or indirect detection (such as epitope tag such as the FLAG epitope, enzyme tag such as horseradish peroxidase). The label can be bound to the antibody, or incorporated into the structure of the antibody.

[0061] A variety of methods can be used to detect the label, depending on the nature of the label and other assay components. For example, the label can be detected while bound to the solid substrate or subsequent to separation from the solid substrate. Labels can be directly detected through optical or electron density, radioactive emissions, nonradiative energy transfers or indirectly detected with antibody conjugates, or streptavidin-biotin conjugates. Methods for detecting the labels are well known in the art.Pharmaceutical Compositions

[0062] Pharmaceutical compositions comprising an anti-CRH antibody (or antigen binding fragment thereof) described herein are also contemplated. In some embodiments, the pharmaceutical composition contains formulation materials for modifying, maintaining or preserving, for example, the pH, osmolarity, viscosity, clarity, color, isotonicity, odor, sterility, stability, rate of dissolution or release, adsorption or penetration of the composition. In such embodiments, suitable formulation materials include, but are not limited to, amino acids (such as glycine, glutamine, asparagine, arginine, proline, methionine or lysine); antimicrobials; antioxidants (such as reducing agents, oxygen / free-radical scavengers, and chelating agents (e.g., ascorbic acid, EDTA, sodium sulfite or sodium hydrogen-sulfite)); buffers (such as borate, bicarbonate, Tris-HCI, citrates, phosphates or other organic acids); bulking agents (such as mannitol or glycine); chelating agents (such as ethylenediamine tetraacetic acid (EDTA)); complexing agents (such as caffeine, polyvinylpyrrolidone, beta-cyclodextrin or hydroxypropyl-beta-cyclodextrin); fillers; monosaccharides; disaccharides; and other carbohydrates (such as glucose, mannose or dextrins); proteins (such as serum albumin, gelatin or immunoglobulins); coloring, flavoring and diluting agents; emulsifying agents; hydrophilic polymers (such as polyvinylpyrrolidone); low molecular weight polypeptides; salt-forming counter-ions (such as sodium); preservatives (such as benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid or hydrogen peroxide); solvents (such as glycerin, propylene glycol or polyethylene glycol); sugar alcohols (such as mannitol or sorbitol); suspending agents; surfactants or wetting agents (such as pluronics, PEG,sorbitan esters, polysorbates such as polysorbate 20, polysorbate, triton, tromethamine, lecithin, cholesterol, tyloxapal); stability enhancing agents (such as sucrose or sorbitol); tonicity enhancing agents (such as alkali metal halides, preferably sodium or potassium chloride, mannitol sorbitol); delivery vehicles; diluents; excipients and / or pharmaceutical adjuvants. See, REMINGTON'S PHARMACEUTICAL SCIENCES, 18" Edition, (A. R. Genrmo, ed.), 1990, Mack Publishing Company.

[0063] Selection of the particular formulation materials described herein may be driven by, for example, the intended route of administration, delivery format and desired dosage. See, for example, REMINGTON'S PHARMACEUTICAL SCIENCES, supra. The primary vehicle or carrier in a pharmaceutical composition may be either aqueous or non-aqueous in nature. For example, a suitable vehicle or carrier may be water for injection, physiological saline solution or artificial cerebrospinal fluid, possibly supplemented with other materials common in compositions for parenteral administration. Neutral buffered saline or saline mixed with serum albumin are further exemplary vehicles. In specific embodiments, pharmaceutical compositions comprise Tris buffer of about pH 7.0-8.5, or acetate buffer of about pH 4.0-5.5, and may further include sorbitol or a suitable substitute therefore. In certain embodiments, the composition may be prepared for storage by mixing the selected composition having the desired degree of purity with optional formulation agents (REMINGTON'S PHARMACEUTICAL SCIENCES, supra) in the form of a lyophilized cake or an aqueous solution. Further, in some embodiments, the antibody or (antigen binding fragment thereof) may be formulated as a lyophilizate using appropriate excipients such as sucrose.

[0064] The pharmaceutical compositions described herein can be selected for parenteral delivery. Alternatively, the compositions may be selected for inhalation or for delivery through the digestive tract, such as orally. Preparation of such pharmaceutically acceptable compositions is within the skill of the art. The formulation components are present preferably in concentrations that are acceptable to the site of administration. In certain embodiments, buffers are used to maintain the composition at physiological pH or at a slightly lower pH, typically within a pH range of from about 5 to about 8.

[0065] When parenteral administration is contemplated, the composition may be provided in the form of a pyrogen-free, parenterally acceptable aqueous solution comprising the desired antibody or fragment in a pharmaceutically acceptable vehicle. A particularly suitable vehicle for parenteral injection is sterile distilled water in which the antibody or fragment is formulated as a sterile, isotonic solution, properly preserved. In certain embodiments, implantable drug delivery devices may be used to introduce the desired antibody (or antigen binding fragment thereof).

[0066] Additional pharmaceutical compositions will be evident to those skilled in the art, including formulations involving antigen binding proteins in sustained- or control led-delivery formulations. Techniques for formulating a variety of other sustained- or controlled-delivery means, such as liposome carriers, bio-erodible microparticles or porous beads and depot injections, are also known to those skilled in the art. See, for example, International Patent Application No. PCT / US93 / 00829, which is incorporated by reference and describescontrolled release of porous polymeric microparticles for delivery of pharmaceutical compositions. Sustained- release preparations may include semipermeable polymer matrices in the form of shaped articles, e.g., films, or microcapsules. Sustained release matrices may include polyesters, hydrogels, polylactides (as disclosed in U.S. Pat. No. 3773919 and European Patent Application Publication No. EP058481, each of which is incorporated by reference), copolymers of L-glutamic acid and gamma ethyl-L-glutamate (Sidman et al., 1983, Biopolymers 2:547-556), poly (2-hydroxyethyl-methacrylate) (Langer et al., 1981, J. Biomed. Mater. Res. 15:167-277 and Langer, 1982, Chem. Tech. 12:98-105), ethylene vinyl acetate (Langer et al., 1981, supra) or poly-D(-)-3- hydroxybutyric acid (European Patent Application Publication No. EP133988). Sustained release compositions may also include liposomes that can be prepared by any of several methods known in the art. See, e.g., Eppstein et al., 1985, Proc. Natl. Acad. Sci. U.S.A. 82:3688-3692; European Patent Application Publication Nos. EP036676; EP088046 and EP143949, incorporated by reference.

[0067] Embodiments of the antibody formulations can further comprise one or more preservatives.

[0068] Administration of the compositions described herein will be via any common route so long as the target tissue is available via that route. The pharmaceutical compositions may be introduced into the subject by any conventional method, e.g., by intravenous, intradermal, intramuscular, intramammary, intraperitoneal, intrathecal, intraocular, retrobulbar, intrapulmonary (e.g., term release); by oral, sublingual, nasal, anal, vaginal, or transdermal delivery, or by surgical implantation at a particular site, e.g., embedded under the splenic capsule, brain, or in the cornea. The treatment may consist of a single dose or a plurality of doses over a period of time. In some embodiments, the composition is delivered via local administration to the brain.

[0069] Therapeutic Methods

[0070] The antibodies (or antigen binding fragments thereof) and pharmaceutical compositions described herein are useful for treating or preventing disorders associated with CRH dysregulation or hypothalamic- pituitary-adrenal (HPA) axis activation. For example, described here are methods of inhibiting the hypothalamic pituitary adrenal (HPA) axis in a subject comprising administering an antibody (or antigen binding fragment thereof) to the subject. Exemplary disorders associated with CRH dysregulation or HPA axis activation include, but are not limited to, anxiety, depression, Alzheimer's Disease, post-traumatic stress disorder, generalized anxiety disorder, major depression, anorexia nervosa, adrenal disorder, metabolic syndrome, type 1 diabetes, sarcopenia, and multiple sclerosis

[0071] The HPA axis includes positive and negative feedback interactions among three endocrine glands: the hypothalamus, the pituitary gland, and the adrenal glands that form the neuroendocrine system. Hormones released by the endocrine glands control reactions to stress, regulation of body processes like digestion, the immune system, mood and emotions, sexuality and energy storage and expenditure. The HPA axis is dysregulated in several psychiatric and neuropsychiatric diseases, as well as in alcoholism and stroke. Examples of HPA axis biomarkers include adrenocorticotropic hormone (ACTH) and cortisol. Cortisol inhibits secretion of corticotropin-releasing hormone (CRH), resulting in feedback inhibition of ACTH secretion. This normal feedbackloop may break down when humans are exposed to chronic stress, and may be an underlying cause of depression.

[0072] Chronic psychological stress and perturbation of the HPA-axis have been associated with many diseases and conditions including, but not limited to, Alzheimer's disease, anxiety disorders, major depression, post-traumatic stress disorder, addiction, metabolic syndrome, osteoporosis and sarcopenia. Further, chronic stress, HPA-axis dysfunction, and elevated cortisol have been implicated as an accelerant with regard to the negative physiological consequences of aging. Treatment of one or more of these disorders / conditions by administering an anti-CRH antibody described herein to a subject in need thereof is specifically contemplated.

[0073] In another aspect, described herein are methods of treating a stress-related disorder in a subject in need thereof, comprising administering to the subject the antibody or antigen binding fragment described herein in an amount effective to treat the disorder. In some embodiments, the stress-related disorder is selected from the group consisting of anxiety, depression, Alzheimer's Disease, post-traumatic stress disorder, generalized anxiety disorder, major depression, anorexia nervosa, adrenal disorder, metabolic syndrome, type 1 diabetes, sarcopenia, and multiple sclerosis.

[0074] In another aspect, described herein are methods of reducing corticotropin releasing hormone, adrenocorticotropin releasing hormone, and / or cortisol in a subject in need thereof comprising administering to the subject an antibody (or antigen binding fragment) described herein in an amount effective to reduce corticotropin releasing hormone, adrenocorticotropin releasing hormone, and / or cortisol in the subject.

[0075] CRH dysregulation has also been associated with the development of obesity in subjects (Vasconcelos et al., Obesity Rev., 25:e13763, 2024). Thus, the use of the antibodies described herein in the context of obesity is specifically contemplated. For example, the disclosure provides methods of treating obesity in a subject in need thereof comprising administering to the subject an antibody (or antigen binding fragment) described herein to the subject. In some embodiments, treatment with an antibody (or antigen binding fragment) described herein results in at least 1%, reduction in total body fat of the subject. In some embodiments, treatment with an antibody (or antigen binding fragment) described herein results in at least 1%, at least 5%, at least 10%, at least 15%, at least 20% reduction or more in total body fat of the subject.

[0076] By "treatment" or "treating” it is meant that at least an amelioration of one or more symptoms associated with CRH dysregulation or hypothalamic-pituitary-adrenal (HPA) axis activation (e.g., stress-related disorders and / or cancer) afflicting the patient, where amelioration is used in a broad sense to refer to at least a reduction in the magnitude of a parameter, e.g., a symptom associated with the disease being treated. As such, treatment also includes situations where a pathological condition, or at least symptoms associated therewith, is completely inhibited, e.g., prevented from happening, or stopped, e.g., terminated, such that patient no longer suffers from the impairment, or at least the symptoms that characterize the impairment. In some instances, "treatment", "treating" and the like refer to obtaining a desired pharmacologic and / or physiologic effect. The effect may be prophylactic in terms of completely or partially preventing a disease or symptom thereof and / ormay be therapeutic in terms of a partial or complete cure for a disease and / or adverse effect attributable to the disease. "Treatment" may be any treatment of a disease in a subject, and includes: (a) preventing the disease from occurring in a subject which may be predisposed to the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., arresting its development; or (c) relieving the disease, i.e., causing regression of the disease. Treatment may result in a variety of different physical manifestations, e.g., modulation in gene expression, increased neurogenesis, rejuvenation of tissue or organs, etc. Treatment of ongoing disease, where the treatment stabilizes or reduces the undesirable clinical symptoms of the patient, occurs in some embodiments. Such treatment may be performed prior to complete loss of function in the affected tissues. The subject therapy may be administered during the symptomatic stage of the disease, and in some cases after the symptomatic stage of the disease. The term "preventing” does not mean that there is 100% inhibition of the disease.

[0077] Chronic stress has been associated with a number of behavioral and physiologic morbidities. Psychological stress contributes to numerous behavioral or psychiatric conditions including, but not limited to, generalized anxiety disorder, panic disorder, major depression, Alzheimer's disease, and post-traumatic stress disorder (PTSD). Chronic stress also has also been shown to have adverse cardiovascular and immunologic effects. CRH is the main coordinator of the HPA stress response and is strongly implicated in animal models of anxiety, addiction, major depression and PTSD. Further, Alzheimer's disease stress and CRH are powerful accelerants of both amyloid and tau pathologies, and CRH drives amyloid pathology in preclinical models of Alzheimer's disease. Thus, the disclosure contemplates a method of treating or preventing anxiety, depression, Alzheimer's Disease, post traumatic stress disorder, generalized anxiety disorder, major depression, anorexia nervosa, post-traumatic stress disorder, adrenal disorder, metabolic syndrome, type 1 diabetes, sarcopenia and multiple sclerosis in a subject in need thereof comprising administering an antibody (or antigen binding fragment) described herein to the subject.

[0078] CRH has been detected in cancer tissues and cell lines and may stimulate cell motility and invasiveness of MCF7 breast cancer cells (Androulidaki et al., Mol. Cancer, 8:30, 2009). Thus, the disclosure contemplates a method of treating cancer in a subject in need thereof comprising administering an antibody (or antigen binding fragment) described herein to the subject. Exemplary cancers include, but are not limited to, esophageal cancer, pancreatic cancer, metastatic pancreatic cancer, metastatic adenocarcinoma of the pancreas, bladder cancer, stomach cancer, fibrotic cancer, glioma, malignant glioma, diffuse intrinsic pontine glioma, recurrent childhood brain neoplasm renal cell carcinoma, clear-cell metastatic renal cell carcinoma, kidney cancer, prostate cancer, metastatic castration resistant prostate cancer, stage IV prostate cancer, metastatic melanoma, melanoma, malignant melanoma, recurrent melanoma of the skin, melanoma brain metastases, stage IIIA skin melanoma; stage IIIB skin melanoma, stage IIIC skin melanoma; stage IV skin melanoma, malignant melanoma of head and neck, lung cancer, non-small cell lung cancer (NSCLC), squamous cell non-small cell lung cancer, breast cancer, recurrent metastatic breast cancer, hepatocellular carcinoma, Hodgkin's lymphoma, follicular lymphoma, non-Hodgkin's lymphoma, advanced B-cell NHL, HL including diffuselarge B-cell lymphoma (DLBCL), multiple myeloma, chronic myeloid leukemia, adult acute myeloid leukemia in remission; adult acute myeloid leukemia with Inv(16)(p13.1q22); CBFB-MYH11; adult acute myeloid leukemia with t(16; 16)(p13.1 ;q22); CBFB-MYH11; adult acute myeloid leukemia with t(8;21)(q22;q22); RUNX1-RUNX1T1; adult acute myeloid leukemia with t(9; 11 )(p22;q23); MLLT3-MLL; adult acute promyelocytic leukemia with t(15; 17)(q22;q 12); PML-RARA; alkylating agent-related acute myeloid leukemia, chronic lymphocytic leukemia, richter's syndrome; Waldenstrom macroglobulinemia, adult glioblastoma; adult gliosarcoma, recurrent glioblastoma, recurrent childhood rhabdomyosarcoma, recurrent Ewing sarcoma / peripheral primitive neuroectodermal tumor, recurrent neuroblastoma; recurrent osteosarcoma, colorectal cancer, MSI positive colorectal cancer; MSI negative colorectal cancer, nasopharyngeal nonkeratinizing carcinoma; recurrent nasopharyngeal undifferentiated carcinoma, cervical adenocarcinoma; cervical adenosquamous carcinoma; cervical squamous cell carcinoma; recurrent cervical carcinoma; stage IVA cervical cancer; stage IVB cervical cancer, anal canal squamous cell carcinoma; metastatic anal canal carcinoma; recurrent anal canal carcinoma, recurrent head and neck cancer; carcinoma, squamous cell of head and neck, head and neck squamous cell carcinoma (HNSCC), ovarian carcinoma, colon cancer, gastric cancer, advanced Gl cancer, gastric adenocarcinoma; gastroesophageal junction adenocarcinoma, bone neoplasms, soft tissue sarcoma; bone sarcoma, thymic carcinoma, urothelial carcinoma, recurrent merkel cell carcinoma; stage III merkel cell carcinoma; stage IV merkel cell carcinoma, myelodysplastic syndrome and recurrent mycosis fungoides and Sezary syndrome.

[0079] In some embodiments, one or more doses of an antibody or antigen binding fragment described herein are administered in an amount and for a time effective to reduce or inhibit corticosterone response to stress, treat or prevent a stress-related disorder, or to treat obesity. For example, one or more administrations of an antibody or antigen binding fragment thereof described herein are optionally carried out over a therapeutic period of, for example, about 1 week to about 24 months (e.g., about 1 month to about 12 months, about 1 month to about 18 months, about 1 month to about 9 months or about 1 month to about 6 months or about 1 month to about 3 months). In some embodiments, a subject is administered one or more doses of an antibody or fragment thereof described herein over a therapeutic period of, for example about 1 month to about 12 months (52 weeks) (e.g., about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, or about 11 months).

[0080] It may be advantageous to administer multiple doses of an antibody or antigen binding fragment described herein at a regular interval, depending on the therapeutic regimen selected for a particular subject. In some embodiments, the antibody or fragment thereof is administered periodically over a time period of one year (12 months, 52 weeks) or less (e.g., 9 months or less, 6 months or less, or 3 months or less). In this regard, the antibody or fragment thereof is administered to the human once every about 3 days, or about 7 days, or 2 weeks, or 3 weeks, or 4 weeks, or 5 weeks, or 6 weeks, or 7 weeks, or 8 weeks, or 9 weeks, or 10 weeks, or 11 weeks, or 12 weeks, or 13 weeks, or 14 weeks, or 15 weeks, or 16 weeks, or 17 weeks, or 18 weeks, or 19 weeks, or 20 weeks, or 21 weeks, or 22 weeks, or 23 weeks, or 6 months, or 12 months.

[0081] In some embodiments, the one or more doses can comprise between about 0.1 to about 50 milligrams (e.g., between about 5 and about 50 milligrams), or about 1 to about 100 milligrams, of antibody (or antigen binding fragment thereof) per kilogram of subject body weight (mg / kg) . For example, the dose may comprise at least about 0.1 mg / kg, at least about 0.5 mg / kg, at least about 1 mg / kg, at least about 2 mg / kg, at least about 3 mg / kg, at least about 4 mg / kg, at least about 5 mg / kg, at least about 6 mg / kg, at least about 7 mg / kg, at least about 8 mg / kg, at least about 9 mg / kg, at least about 10 mg / kg, at least about 20 mg / kg, at least about 25 mg / kg, at least about 26 mg / kg, at least about 27 mg / kg, at least about 28 mg / kg, at least about 29 mg / kg, at least about 30 mg / kg, at least about 31 mg / kg, at least about 32 mg / kg, at least about 33 mg / kg, at least about 34 mg / kg, at least about 35 mg / kg, at least about 36 mg / kg, at least about 37 mg / kg, at least about 38 mg / kg, at least about 39 mg / kg, at least about 40 mg / kg, at least about 41 mg / kg, at least about 42 mg / kg, at least about 43 mg / kg, at least about 44 mg / kg, at least about 45 mg / kg, at least about 46 mg / kg, at least about 47 mg / kg, at least about 48 mg / kg, at least about 49 mg / kg, at least about 50 mg / kg, at least about 55 mg / kg, at least about 60 mg / kg, at least about 65 mg / kg, at least about 70 mg / kg, at least about 75 mg / kg, at least about 80 mg / kg, at least about 85 mg / kg, at least about 90 mg / kg, at least about 95 mg / kg, or up to about 100 mg / kg. Ranges between any and all of these endpoints are also contemplated, e.g., about 1 mg / kg to about 3 mg / kg, about 1 mg / kg to about 5 mg / kg, about 1 mg / kg to about 8 mg / kb, about 3 mg / kg to about 8 mg. kg, about 1 mg / kg to about 10 mg / kg, about 1 mg / kg to about 20 mg / kg, about 1 mg / kg to about 40 mg / kg, about 5 mg / kg to about 30 mg / kg, or about 5 mg / kg to about 20 mg / kg.

[0082] Combination therapy

[0083] In various embodiments, the antibodies (or antigen binding fragments thereof) described herein are administered in combination with an additional therapeutic useful for treating a condition or disorder associated with HPA axis activation described herein. In some embodiments, the additional therapeutic is Hydrocortisone rescue therapy, Glucocorticoid supplementation and / or a GLP-1 agonist (e.g., a glutide). In some embodiments, the additional therapeutic is a GLP-1 agonist. In some embodiments, the GLP-1 agonist is dulaglutide, exenatide, exenatide extended release, liraglutide, lixisenatide, insulin degludec / liraglutide, insulin glargine / lixisenatide, semaglutide, and tirzepatide.

[0084] Concurrent administration of two therapeutic agents does not require that the agents be administered at the same time or by the same route, as long as there is an overlap in the time period during which the agents are exerting their therapeutic effect. Simultaneous or sequential administration is contemplated, as is administration on different days or weeks.

[0085] It is contemplated that the anti-CRH antibody (or antigen binding fragment) and the additional therapeutic may be given simultaneously, in the same formulation. It is further contemplated that the anti-CRH antibody (or antigen binding fragment) and the additional therapeutic are administered in a separate formulation and administered concurrently, with concurrently referring to agents given within 30 minutes of each other.

[0086] In another aspect, the anti-CRH antibody (or antigen binding fragment thereof) is administered prior to administration of the additional therapeutic. Prior administration refers to administration of the anti-CRH antibody within the range of one week prior to treatment with the additional therapeutic up to 30 minutes before administration of the additional therapeutic. It is further contemplated that the anti-CRH antibody is administered subsequent to administration of the additional therapeutic. Subsequent administration is meant to describe administration from 30 minutes after administration of the additional therapeutic to up to one week after administration of the additional therapeutic.

[0087] Kits

[0088] Once a pharmaceutical composition has been formulated, it may be stored in sterile vials as a solution, suspension, gel, emulsion, solid, crystal, or as a dehydrated or lyophilized powder. Such formulations may be stored either in a ready-to-use form or in a form (e.g., lyophilized) that is reconstituted prior to administration. The invention also provides kits for producing a single-dose administration unit. The kits of the disclosure may each contain both a first container having a dried protein and a second container having an aqueous formulation. In certain embodiments, kits containing single and multi-chambered pre-filled syringes (e.g., liquid syringes and lyosyringes) are provided.

[0089] The following Examples are provided to further illustrate aspects of the disclosure, and are not meant to constrain the disclosure to any particular application or theory of operation.EXAMPLESExample 1 - Generation of Humanized CRH Antibodies

[0090] Three humanized variable heavy and three humanized variable light regions were designed using a 3D modeling algorithm. A T20 Analyzer score (Gao et al., BMC Biotechnology, 13:55, 2013) was used to evaluate the humanness of the framework regions for all humanized sequences. The T20 Analyzer score of framework humanness ranged from 86 - 97, which improved upon the parental VH and VL framework score.

[0091] Humanized CRH antibodies were formatted as hlgG1 N297A. Expression constructs for nine (9) different humanized CRH antibodies (each combination of three humanized variable heavy chains and three humanized variable light chains) were cloned into a high expression mammalian vector, and the DNA sequences of the gene insert were confirmed. Each DNA construct was scaled up for transfection, and the DNA sequence was confirmed after DNA scale-up. A 0.03 liter transient production was completed in CHO cells for each antibody. The proteins were purified by Protein A chromatography, yielding 2.31 mg to 3.33 mg antibody.

[0092] CE-SDS and mass spectrometry analysis was performed to confirm structural stability. The observed molecular weights for the IgGs were within the expected range.Example 2 - Affinity of Humanized CRH Antibodies

[0093] The antibodies generated in Example 1 have a high affinity for CRH. Binding assessment by ELISA was performed. First conditions were scouted and confirmed using a subset of IgG (data not shown). Testing all (9) IgG, ELISA EC50 values were calculated, ranging from 39 nM to greater than 2000 nM.

[0094] Binding kinetics of the humanized CRH antibodies were also quantified using an Octet® Bio-layer interferometry system assay. Briefly, biotinylated CRF peptide was immobilized as a Ligand and all nine (9) humanized CRH antibodies were tested as the Analyte. Binding experiments were performed on Octet HTX at 25C. Biotin-CRF peptide (10 pg / mL) was loaded onto Streptavidin biosensors (SAX). Loaded sensors were dipped into IgG samples, prepared as serial dilutions (Top concentration of 1000 nM with three-fold dilutions and seven concentrations in total). Kinetic constants were calculated using a monovalent (1 :1) binding model. Calculated KD values of the humanized CRH antibodies range from 732 pM to 58 nM.

[0095] Humanized CRH antibodies were further optimized by introducing point mutations to disrupt or eliminate amino acid sequence motif liabilities known to undergo chemical degradation during large scale manufacturing processes. A first batch of 40 optimized humanized CRH antibodies were formatted as hlgG1 N297A, cloned and expressed as described above.

[0096] Binding kinetics of the optimized humanized CRH antibodies were quantified using an OctetD Biolayer interferometry system assay. Briefly, biotinylated CRF peptide was immobilized as a Ligand and all forty (40) optimized humanized CRH antibodies were tested as the Analyte. Binding experiments were performed on Octet HTX at 25C. Biotin-CRF peptide (10 pg / mL) was loaded onto Streptavidin biosensors (SAX). Loaded sensors were dipped into IgG samples, prepared as serial dilutions (Top concentration of 333 nM with three-fold dilutions and seven concentrations in total). Kinetic constants were calculated using a monovalent (1 :1) binding model. Calculated KD values of the optimized humanized CRH antibodies range from 330 pM to 3.4 nM.

[0097] The CRH binding properties of humanized CRH antibodies were further optimized by introducing point mutations within the antibody combining site. Expression constructs of Humanized CRH antibodies carrying new point mutations were subcloned into the mammalian expression vector pCDNA3.1 with either zeocin or hygromycin selection marker. Each DNA construct was scaled up for transfection, and the DNA sequence was confirmed after DNA scale-up. A small-scale transient production was completed in 293T cells for each antibody using the PEI transfection method. Antibody yield and stability were assayed by western blotting with or without reducing reagent. Initial binding assessments were performed using the ELISA binding assay. Humanized CRH antibody mutants that achieved a predetermined binding threshold activity were additionally transiently produced in a large culture format and purified using Protein A chromatography for kinetic analysis using an OctetD Biolayer interferometry system assay.

[0098] ELISA binding data was obtained by coating ELISA plates with 10 nM CRH to capture the humanized CRH antibody mutants (Futch et al., JEM, 216(11), 2019). Optical density (O.D.) ratios were recorded and used to calculate a fold change in binding activity for all newly generated humanized CRH antibody mutant variants.Multiple humanized CRH antibodies mutant variants displayed greater than a ten (10)-fold increase in CRH binding activity over the initial nine (9), non-optimized humanized CRH antibodies.n.d.: not determined (insufficient binding was observed)Table 2.Sequence Table

Claims

What is claimed is:1 . An antibody or antigen-binding fragment thereof comprising a set of 6 CDRs, wherein the CDRs comprise:(a) a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 1 (DSTMX), SEQ ID NO: 2 (DSTIX) or DSTXX;(b) a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 10 (XIHPDXGGTXYXQKFXG) or SEQ ID NO: 239 (XIHPDSGGTIYNQKFQG);(c) a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 29 (XGFAY), SEQ ID NO: 30 (XPFAY), SEQ ID NO: 31 (NGXAY), SEQ ID NO: 32 (NGFXY), or SEQ ID NO: 33 (TXFAY);(d) a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 46 (RSSXSLLGSXXXTXLX);(e) a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 62 (XVSNKXX); and(f) aCDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 70 (SQXXXHXPX).

2. The antibody of claim 1, wherein the CDR-H1 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 3-9.

3. The antibody of claim 1 or claim 2, wherein the CDR-H2 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 11-28.

4. The antibody of any one of claims 1-3, wherein the CDR-H3 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 34-45.

5. The antibody of any one of claims 1-4, wherein the CDR-L1 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 47-61 .

6. The antibody of any one of claims 1-5, wherein the CDR-L2 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 63-69.

7. The antibody of any one of claims 1-6, wherein the CDR-L3 comprises the amino acid sequence set forth in any one of SEQ ID NOs: 71-85.

8. The antibody of any one of claims 1-7, comprising a heavy chain variable amino acid sequence that is at least 80% identical to an amino acid sequence set forth in any one of SEQ ID NOs: 163-193.

9. The antibody of any one of claims 1-8, comprising a light chain variable amino acid sequence that is at least 80% identical to an amino acid sequence set forth in any one of SEQ ID NOs: 194-238.

10. The antibody of any one of claims 1-9, comprising a heavy chain variable region amino acid sequence set forth in any one of SEQ ID NOs: 163-193.

11. The antibody of any one of claims 1-10, comprising a light chain variable region amino acid sequence set forth in any one of SEQ ID NOs: 194-238.

12. The antibody of any one of claims 1-11, comprising a heavy chain amino acid sequence that is at least 80% identical to an amino acid sequence set forth in any one of SEQ ID NOs: 89-118.

13. The antibody of any one of claims 1-12 comprising a light chain amino acid sequence that is at least 80% identical to an amino acid sequence set forth in any one of SEQ ID NOs: 119-163.

14. The antibody of any one of claims 1-13, comprising a heavy chain amino acid sequence set forth in any one of SEQ ID NOs: 89-118.

15. The antibody of any one of claims 1-14, comprising a light chain amino acid sequence set forth in any one of SEQ ID NOs: 119-163.

16. The antibody of any one of claims 1-15, comprising a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 164 and a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 234 or SEQ ID NO: 236.

17. The antibody of any one of claims 1-15, comprising a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 175 and a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 234, or SEQ ID NO: 236.

18. The antibody of any one of claims 1-15, comprising a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 186 and a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 233, SEQ ID NO: 234, SEQ ID NO: 235, or SEQ ID NO: 236.

19. The antibody of any one of claims 1-15, comprising a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 193 and a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 234 or SEQ ID NO: 236.

20. The antibody of any one of claims 1-19, that is a humanized antibody.

21. The antibody of any one of claims 1-20, that is an IgG.

22. The antibody of any one of claims 1-21, comprising an Fc region.

23. The antibody of claim 22, wherein the Fc region comprises a substitution that reduces effector function as compared to effector function of the antibody with a wild-type Fc.

24. An isolated antibody or antigen binding fragment thereof that binds to corticotropin releasing hormone (CRH) comprising three heavy chain complementarity determining regions (CDRs) (CDR-H1, CDR-H2,and CDR-H3) and three light chain complementarity determining regions (CDRs) (CDR-L1, CDR-L2, and CDR- L3), wherein the CDR-H1, CDR-H2, and CDR-H3 are present in a heavy chain variable domain (VH) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 163-193 and wherein the CDR-L1, CDR-L2, and CDR-L3 are present in a light chain variable domain (VL) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 194-238.

25. The isolated antibody or antigen binding fragment of claim 24, wherein: the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to Kabat; the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to Chothia; the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to IMGT; the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to AbM; the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to Contact; or the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to Honneger (AHo).

26. A method of treating a stress-related disorder in a subject in need thereof, comprising administering to the subject the antibody or antigen binding fragment of any one of claims 1-25 in an amount effective to treat the disorder.

27. The method of claim 26, wherein the stress-related disorder is selected from the group consisting of anxiety, depression, Alzheimer's Disease, post-traumatic stress disorder, generalized anxiety disorder, major depression, anorexia nervosa, adrenal disorder, metabolic syndrome, type 1 diabetes, sarcopenia, and multiple sclerosis.

28. A method of inhibiting the hypothalamic pituitary adrenal (HPA) axis in a subject in need thereof comprising administering to the subject the antibody of any one of claims 1-25.

29. A method of reducing corticotropin releasing hormone, adrenocorticotropin releasing hormone, and / or cortisol in a subject in need thereof, the method comprising administering to the subject the antibody of any one of claims 1-25.

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