Compositions and methods for treatment of thyroid eye disease

Abstract

To provide antibodies and compositions against IGF-1R and uses thereof.SOLUTION: Provided is an antibody comprising a light chain, a heavy chain, and an Fc region, wherein the light chain comprises a light chain variable region including LCDR1, LCDR2, and LCDR3 having specific amino acid sequences; the heavy chain comprises a heavy chain variable region including HCDR1, HCDR2, and HCDR3 having specific amino acid sequences; and the Fc region comprises substitutions of M252Y, S254T, and T256E numbered according to the Kabat numbering system EU index.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application Nos. 63 / 091,839, filed October 14, 2020, 63 / 201,978, filed May 21, 2021, 63 / 260,130, filed August 10, 2021, and 63 / 261,742, filed September 28, 2021, each of which is incorporated by reference in its entirety. [Background technology]

[0002] Thyroid-associated eye disease (TAO), also known as thyroid eye disease (TED), Graves' eye disease or ophthalmopathy (GO), thyrotoxic exophthalmos, hypothyroid eye disease, and several other terms, is an eye disease associated with thyroid dysfunction. TAO is divided into two types. Active TAO, which usually lasts 1 to 3 years, is characterized by a progressive autoimmune / inflammatory response in the soft tissues of the orbit. Active TAO is responsible for swelling and remodeling of the ocular soft tissues. The autoimmune / inflammatory response in active TAO resolves spontaneously, and the condition transitions to inactive TAO. Inactive TAO is a term used to describe the long-term / persistent sequelae of active TAO. The cause of TAO is unknown. TAO is usually associated with Grave's hyperthyroidism, but it can also occur as part of other autoimmune conditions that affect the thyroid gland and produce pathology in the orbit and periorbital tissues, and, rarely, in the pretibial skin (pretibial myxedema) or digits (thyroid clubbed acromyopathy). TAO is an autoimmune ophthalmopathy that primarily affects the orbit and periocular soft tissues, with the eye and vision secondarily affected. In TAO, the eyeball is pushed forward (bulged) from the orbit (a phenomenon called proptosis or exophthalmos) as a result of inflammation and swelling of the orbital soft tissues, primarily the ocular muscles and fat. While most cases of TAO do not result in blindness, the condition can cause sight-threatening lagophthalmos, troublesome diplopia (double vision), and compressive hypothyroid optic neuropathy. TAO may precede, occur concurrently with, or follow the systemic complications of thyroid insufficiency. Ocular manifestations of TAO include upper eyelid retraction, eyelid lag, swelling, redness (erythema), conjunctivitis, and ocular bulging (proptosis or exophthalmos), chemosis, periorbital edema, and altered ocular motility, with significant functional, social, and cosmetic impacts. Many signs and symptoms of TAO, including exophthalmos and ocular congestion, result from expansion of orbital adipose tissue and periocular muscles. Adipose tissue volume is due, in part, to adipocyte growth (adipogenesis) within the orbital fat.Accumulation of hydrophilic glycosaminoglycans (mainly hyaluronic acid) within the orbital adipose tissue and within the perimuscular connective tissue between extraocular muscle fibers further expands the adipose compartment and the extraocular muscle body. Hyaluronic acid is produced by fibroblasts within the orbital fat and extraocular muscles, and in vitro hyaluronic acid synthesis is stimulated by several cytokines and growth factors, including IL-1β, interferon-γ, platelet-derived growth factor, thyroid-stimulating hormone (TSH), and insulin-like growth factor (IGF-1).

[0003] Antibodies that activate the insulin-like growth factor I receptor (IGF-IR) have also been detected in, and suggested to be involved in, active TAO. Without being bound by any theory, it is believed that TSHR and IGF-IR form a physical and functional complex in orbital fibroblasts, and blocking IGF-IR appears to attenuate both IGF-I and TSH-dependent signaling. It has been suggested that blocking IGF-IR with antibody antagonists may reduce both TSHR and IGF-I-dependent signaling, thus preventing the pathological activity of autoantibodies that act as agonists at either receptor. There are.

[0004] IGF-IR is a ubiquitously expressed heterotetrameric protein involved in the regulation of growth and metabolic functions in many cell types. IGF-IR is a tyrosine kinase receptor containing two subunits. IGF-IRα contains the ligand-binding domain, while IGF-IRβ is involved in signal transduction and contains a tyrosine phosphorylation site.

[0005] Current treatments for hyperthyroidism due to Graves' disease are incomplete due to the lack of therapies targeting the specific pathogenic autoimmune mechanisms underlying the disease. Treatment of moderate to severe active TAO is even more complex. Although the pathogenesis of TAO has become better understood in recent years, TAO remains a therapeutic challenge. No medications are approved for the treatment of active TAO. Patients with moderate to severe active TAO have been treated with intravenous glucocorticoids (ivGCs) and oral glucocorticoids, but satisfactory results have rarely been achieved. Partial responses are frequent, and relapse (rebound) after discontinuation of medication is not uncommon. Adverse events are real, and many patients ultimately require rehabilitative surgery when their condition progresses to inactive TAO. Therefore, there remains a need to provide alternative treatments for TAO and its associated symptoms. Summary of the Invention [Means for solving the problem]

[0006] Embodiments generally relate to IGF-1R antibodies and antigen-binding fragments thereof. Certain IGF-1R antibodies and antigen-binding fragments inhibit IGF-1R function or block the biological function of IGF-1-mediated IGF-1R signaling. Furthermore, the present invention generally relates to methods for treating thyroid-associated eye disease (TAO), also known as thyroid eye disease (TED), Graves' eye disease or ophthalmopathy (GO), thyrotoxic exophthalmopathy, hypothyroid eye disease, and other thyroid eye disorders associated with IGF-1R signaling.

[0007] In some embodiments, an antibody or antigen-binding fragment thereof is provided, comprising a sequence set forth herein. In some embodiments, the antibody comprises a VL sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 79, or 86; and a VH sequence set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 80, or 83. In some embodiments, the antibody comprises an LCDR sequence set forth in SEQ ID NO: 17, 18, 19, 23, 24, 25, 29, 30, 31, 35, 36, 37, 41, 42, 43, 47, 48, 49, 53, 54, 55, 59, 60, 61, or 81 and an HCDR sequence set forth in SEQ ID NO: 20, 21, 22, 26, 27, 28, 32, 33, 34, 38, 39, 40, 44, 45, 46, 50, 51, 52, 56, 57, 58, 62, 63, or 64; or any combination or variant thereof.

[0008] In some embodiments, the antibody or antigen-binding fragment thereof comprises a V sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 79, or 86. L In some embodiments, the antibody or antigen-binding fragment thereof comprises a V peptide set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 80, or 83, or a variant of any of these. H peptide, or a variant of any of these.

[0009] In some embodiments, the antibody or antibody fragment comprises (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence is the amino acid sequence of SEQ ID NO: 20, 26, 32, 38, 44, 50, or 56; the heavy chain CDR2 sequence is the amino acid sequence of SEQ ID NO: 21, 27, 33, 39, 45, 51, or 57; and the heavy chain CDR3 sequence is the amino acid sequence of SEQ ID NO: 22, 28, 34, 40, 46, 52, or 58. and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 17, 23, 29, 35, 41, 47, or 53; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 18, 24, 30, 36, 42, 48, or 54; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 19, 25, 31, 37, 43, 49, 55, or 81; or a variant of any of the foregoing.

[0010] In some embodiments, the antibody or antibody fragment comprises: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 20; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 21; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 22; or any variant of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 17; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 18; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 19; or any variant of the foregoing.

[0011] In some embodiments, the antibody or antibody fragment comprises: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 26; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 27; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 28; or any variant of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 23; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 24; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 25; or any variant of the foregoing.

[0012] In some embodiments, the antibody or antibody fragment comprises: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 32; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 33; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 34; or any variant of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 29; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 30; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 31; or any variant of the foregoing.

[0013] In some embodiments, the antibody or antibody fragment comprises: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 38; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 39; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 40; or any variant of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 35; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 36; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 37; or any variant of the foregoing.

[0014] In some embodiments, the antibody or antibody fragment comprises: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 44; the heavy chain CDR2 has the amino acid sequence of SEQ ID NO: 45; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 46; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 41; The light chain variable region has the sequence of SEQ ID NO: 42; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 43; or a variant of any of the foregoing.

[0015] In some embodiments, the antibody or antibody fragment comprises: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 50; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 51; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 52; or any variant of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 47; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 48; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 49; or any variant of the foregoing.

[0016] In some embodiments, the antibody or antibody fragment comprises: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 56; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 57; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 58; or any variant of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 53; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 54; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 55; or any variant of the foregoing.

[0017] In some embodiments, the antibody or antibody fragment comprises: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 62; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 63; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 64; or any variant of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 59; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 60; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 61; or any variant of the foregoing.

[0018] In some embodiments, the antibody or antibody fragment comprises: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 38; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 39; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 40; or any variant of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 35; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 36; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 81; or any variant of the foregoing.

[0019] In some embodiments, the antibody comprises a V sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 79, or 86. L In some embodiments, the antibody comprises a V sequence set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 80, or 83, or a variant thereof. H sequences, or variants thereof.

[0020] In some embodiments, the antibody comprises the sequence of SEQ ID NO: 65-72, 78, 82, or 85, or a variant thereof.

[0021] In some embodiments, the antibody comprises a light chain having the amino acid sequence of SEQ ID NO: 3 and a heavy chain having the amino acid sequence of SEQ ID NO: 83. In some embodiments, the antibody comprises a light chain variable region having the amino acid sequence of SEQ ID NO: 13 and a heavy chain having the amino acid sequence of SEQ ID NO: 14. The heavy chain variable region comprises:

[0022] In some embodiments, the antibody comprises a light chain having the amino acid sequence of SEQ ID NO:93 and a heavy chain amino acid sequence of SEQ ID NO:92.

[0023] In some embodiments, the antibody comprises a light chain having the amino acid sequence of SEQ ID NO:93 and a heavy chain amino acid sequence of SEQ ID NO:94.

[0024] In some embodiments, the antibody comprises a light chain having the amino acid sequence of SEQ ID NO:93 and a heavy chain amino acid sequence of SEQ ID NO:95.

[0025] In some embodiments, variants of any of the antibodies provided herein are provided, provided that the CDRs are constant compared to the parent (non-variant) sequences provided herein.

[0026] In some embodiments, the antibody comprises an Fc region. In some embodiments, the Fc region is as set forth in SEQ ID NOs: 75-77, 84, 87, 88, 89, or 90. In some embodiments, the Fc region is as set forth in SEQ ID NO: 75. In some embodiments, the Fc region is as set forth in SEQ ID NO: 76. In some embodiments, the Fc region is as set forth in SEQ ID NO: 77. In some embodiments, the Fc region is as set forth in SEQ ID NO: 84. In some embodiments, the Fc region is as set forth in SEQ ID NO: 87. In some embodiments, the Fc region is as set forth in SEQ ID NO: 88. In some embodiments, the Fc region is as set forth in SEQ ID NO: 89. In some embodiments, the Fc region is as set forth in SEQ ID NO: 90.

[0027] In some embodiments, a pharmaceutical composition comprising an antibody provided herein is provided.

[0028] In some embodiments, provided are methods for treating or reducing the severity of thyroid-associated eye disease (TAO) or a symptom thereof, the method comprising administering to a subject an antibody provided herein, or a pharmaceutical composition comprising the antibody.

[0029] In some embodiments, provided are methods for treating thyroid eye disease in a subject, the method comprising administering to the subject an antibody provided herein or a pharmaceutical composition comprising the antibody.

[0030] In some embodiments, provided is a method for reducing the Clinical Activity Score (CAS) of Thyroid Associated Eye Disease (TAO) in a subject, the method comprising administering to the subject an antibody provided herein or a pharmaceutical composition comprising the antibody.

[0031] In some embodiments, provided are methods of a) reducing exophthalmos by at least 2 mm and b) lowering Clinical Activity Score (CAS) in a subject with thyroid-associated eye disease (TAO), the method comprising administering to the subject an antibody provided herein or a pharmaceutical composition comprising the antibody.

[0032] In some embodiments, there is provided a method for treating thyroid-associated eye disease (TAO) or reducing the severity of TAO in a subject, the method comprising administering to the subject an antibody provided herein or a pharmaceutical composition comprising the antibody, wherein treatment with the antibody (i) reduces exophthalmos in the eye by at least 2 mm; (ii) is not accompanied by a worsening of the other (i.e., fellow) eye by 2 mm or more; and (iii) reduces CAS in the subject by (1) or The method provides the above, wherein the concentration of the ion channel is reduced to zero (0).

[0033] In some embodiments, provided are methods for improving quality of life in a subject with thyroid-associated eye disease (TAO, also known as Graves' eye disease / Graves' ophthalmopathy), the method comprising administering to the subject an antibody provided herein or a pharmaceutical composition comprising the antibody.

[0034] In some embodiments, provided are methods for treating or reducing the severity of diplopia in a subject with thyroid-associated eye disease (TAO), the method comprising administering to the subject an antibody provided herein or a pharmaceutical composition comprising the antibody.

[0035] In some embodiments, provided is a method for increasing internalization of IGF-1R in a cell, the method comprising contacting the cell with an antibody provided herein or a pharmaceutical composition comprising the antibody.

[0036] In some embodiments, provided are methods for inhibiting IGF-1 stimulated receptor phosphorylation in a cell, the method comprising contacting the cell with an antibody provided herein or a pharmaceutical composition comprising the antibody.

[0037] In some embodiments, methods are provided for treating thyroid eye disease in a subject, the methods comprising treating the subject with an antibody provided herein or a pharmaceutical composition comprising the antibody, wherein the antibody has a serum concentration in the subject of at least or about 70 μg / mL, 75 μg / mL, 80 μg / mL, 85 μg / mL, 90 μg / mL, 95 μg / mL, 100 μg / mL, or 105 μg / mL at least 1, 2, or 3 weeks after administration.

[0038] In some embodiments, provided are methods for inhibiting IGF-1-induced receptor autophosphorylation in a cell by at least 95%, 96%, 97%, 98%, or 99%, or 100%, the method comprising contacting the cell with an antibody provided herein or a pharmaceutical composition comprising the antibody.

[0039] In some embodiments, any of the methods provided herein is provided in which the antibody, or antigen-binding fragment thereof, is administered in a pharmaceutical composition further comprising a pharmaceutically acceptable diluent, excipient, or carrier. In some embodiments, the pharmaceutical composition further comprises one or more pharmaceutically active compounds for the treatment of TAO. In some embodiments, the pharmaceutical composition further comprises a corticosteroid; rituximab or other anti-CD20 antibody; tocilizumab or other anti-IL-6 antibody; or selenium, infliximab, or other anti-TNF alpha antibody, or a thyroid-stimulating hormone receptor (TSHR) inhibitor. [Brief explanation of the drawings]

[0040] [Figure 1] 1 shows NHP (non-human primate) serum concentrations of various antibodies and embodiments provided herein.

[0041] [Figure 2] 1 illustrates various properties of the antibodies provided herein.

[0042] [Figure 3] 1 illustrates various properties of the antibodies provided herein.

[0043] [Figure 4] 1 illustrates various properties of the antibodies provided herein.

[0044] [Figure 5] 1 illustrates various properties of the antibodies provided herein.

[0045] [Figure 6] 1 illustrates various properties of the antibodies provided herein.

[0046] [Figure 7] 1 illustrates various properties of the antibodies provided herein.

[0047] [Figure 8] 1 illustrates various properties of the antibodies provided herein. DETAILED DESCRIPTION OF THE INVENTION

[0048] Provided herein are antibodies that bind to and regulate the activity of IGF-1R. The antibodies can be used, for example, to treat thyroid eye disease.

[0049] As used herein, "thyroid-associated eye disease" (TAO), "thyroid eye disease" (TED), "Graves' eye disease," or "Graves' ophthalmopathy" (GO) refer to the same disorder or condition and are used interchangeably. They all refer to an inflammatory orbital pathology associated with several autoimmune thyroid disorders, most commonly "Graves' disease" (GD), but occasionally other conditions, such as Hashimoto's thyroiditis.

[0050] The terms "proptosis" and "exophthalmos" (also known as exophthalmos, exophthalmia, or exorbitism) refer to the forward protrusion, displacement, bulging, or protrusion of an organ. As used herein, the term refers to the forward protrusion, displacement, bulging, or protrusion of the eye from the orbit. Proptosis and exophthalmos are considered by those skilled in the art to have the same meaning and are often used interchangeably, although some believe there are subtle differences in their meaning. Exophthalmos is used by some to mean severe bulging of the eye; or to mean endocrine-related bulging of the eye. Still others use the term exophthalmos to describe bulging associated with the eye, for example, in subjects with TAO (TED or GO).

[0051] As used herein, "proptosis" and "exophthalmos" are used interchangeably and refer to the forward projection, displacement, bulging, or protrusion of the eye from the orbit. Because the orbit is a rigid bony structure with only an anterior opening for expansion, any increase in orbital soft tissue content arising from the lateral or posterior regions will cause the eye to be expelled forward. Proptosis or exophthalmos can be the result of several disease processes, including infection, inflammation, tumor, trauma, metastasis, endocrine lesions, vascular disease, and extraorbital pathologies. TAO (TED or GO) is now recognized as the most common cause of exophthalmos in adults. Exophthalmos can be either bilateral, as is often seen with TAO (TED or GO), or unilateral (as is often seen with orbital tumors).

[0052] Measurement of the degree of exophthalmos can be performed, for example, using an exophthalmometer, an instrument used to measure the forward mobility of the eye. This device allows measurement of the distance from the lateral orbital rim to the anterior cornea. Computed tomography (CT) scanning and magnetic resonance imaging (MRI) can also be used to measure the degree of exophthalmos or proptosis. CT scanning is an excellent imaging technique for diagnosing TAO. It allows visualization of enlarged extraocular muscles. In addition, CT scans provide the surgeon or clinician with a depiction of the bony anatomy of the orbit when orbital decompression is necessary. MRI, with its multiplanar and inherent contrast capabilities, provides excellent imaging of the orbital volume without the radiation exposure associated with CT scan studies. While MRI provides better imaging of the optic nerve, orbital fat, and extraocular muscles, CT scans provide a better view of the bony structures of the orbit. Orbital ultrasound can also be used to diagnose and evaluate TAO because it can be performed quickly and reliably. The high reflectivity and hypertrophy of the extraocular muscles are easily measured, and serial ultrasound imaging can also be used to monitor the progression or safety of eye disease. Based on currently available technology or technology that will become available in the future, those skilled in the art can determine the best means for diagnosing and investigating proptosis or exophthalmos.

[0053] As used herein, the term "antibody" refers to any form of antibody that exhibits the desired biological activity. Thus, the term is used in the broadest sense and specifically covers, but is not limited to, monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), humanized, fully human antibodies, chimeric antibodies, and camelized single domain antibodies. A "parent antibody" is an antibody obtained by exposing the immune system to an antigen before modifying the antibody for its intended use, e.g., humanizing the antibody for use as a human therapeutic antibody.

[0054] As used herein, unless otherwise specified, "antibody fragment" or "antigen-binding fragment" refers to an antigen-binding fragment of an antibody, i.e., an antibody fragment that retains the ability to specifically bind to the antigen bound by the full-length antibody, e.g., a fragment that retains one or more CDR regions. Examples of antibody-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules, e.g., sc-Fv; nanobodies formed from antibody fragments, and multispecific antibodies.

[0055] A "Fab fragment" is composed of one light chain and the CH1 and variable regions of one heavy chain. The heavy chain of a Fab molecule cannot form disulfide bonds with another heavy chain molecule.

[0056] The "Fc" region is the C H 1 and C H It contains two heavy chain fragments containing two domains, held together by two or more disulfide bonds and by hydrophobic interactions of the CH3 domains.

[0057] In some embodiments, the antibodies or antigen-binding fragments herein comprise an Fc region. In some embodiments, the Fc region comprises a mutation that extends the half-life of the antibody when bound to the Fc region. In some embodiments, the Fc region comprises S228P, L235E, M252Y, S254T, T256E, M428L, N434S, L234F, P331S mutations, or any combination thereof. In some embodiments, the Fc region comprises M252Y, S254T, and T256E mutations. A non-limiting example of an Fc region comprising M252Y, S254T, and T256E mutations (collectively, "YTE mutations") can be found in the sequence of SEQ ID NO: 89. In some embodiments, an Fc region comprising a YTE mutation comprises the sequence of SEQ ID NO: 90, which differs from SEQ ID NO: 89 in the presence of a C-terminal lysine (K) residue. The numbering of the Fc region can follow the Kabat numbering system for Fc regions.

[0058] In some embodiments, the Fc region comprises S228P and L235E mutations. In some embodiments, the antibody comprises L234F, L235E, and P331S mutations. In some embodiments, the Fc region comprises M252Y, S254T, T256E, S228P, and L235E mutations. In some embodiments, the Fc region comprises S228P, L23 In some embodiments, the Fc region comprises the L234F, L235E, P331S, M252Y, S254T, and T256E mutations. In some embodiments, the Fc region comprises the M428L and N434S mutations. In some embodiments, the Fc region comprises the L234F, L235E, P331S, M252Y, S254T, and T256E mutations. Mutations in the Fc region are also described in US2007041972A1, EP2235059B1, U.S. Patent No. 8,394,925, and Mueller et al., Mol Immunol 1997 Apr;34(6):441-52, each of which is incorporated by reference in its entirety. Numbering referenced herein refers to the Kabat numbering system for the Fc region.

[0059] In some embodiments, the Fc region comprises a sequence selected from the following: APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 75); APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 76); or APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 77); or ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 84) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 87) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 88) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 89) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 90)

[0060] A "Fab' fragment" consists of one light chain and one V H Domain and C H 1 domain, and C H 1 and C H and a portion or fragment of one heavy chain, which also contains the region between the two domains, such that an interchain disulfide bond can form between the two heavy chains of two Fab' fragments to form an F(ab')2 molecule.

[0061] "F(ab')2 fragment" refers to a fragment consisting of two light chains and a CH 1 and C H 2 A F(ab')2 fragment contains two heavy chains containing a portion of the constant region between the heavy chains, such that interchain disulfide bonds are formed between the two heavy chains. Thus, an F(ab')2 fragment is composed of two Fab' fragments held together by disulfide bonds between the two heavy chains.

[0062] The "Fv region" comprises the variable regions from both the heavy and light chains, but lacks the constant regions.

[0063] The term "single-chain Fv" or "scFv" antibody refers to the V of an antibody. H and V L Fv refers to an antibody fragment comprising V domains, wherein these domains are present in a single polypeptide chain. Generally, an Fv polypeptide comprises a V domain that enables the scFv to form the desired structure for antigen binding. H and V L The scFv domains further comprise a polypeptide linker between them. For a review of scFvs, see, e.g., Pluckthun (1994) The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315. See also International Patent Application Publication No. 88 / 01649 and U.S. Patent Nos. 4,946,778 and 5,260,203.

[0064] "Domain antibodies" refer to antibodies that contain only the variable region of a heavy chain or the variable region of a light chain. It is an immunoglobulin fragment that is functional in nature. In some cases, it may contain two or more V H The two V domains of a bivalent domain antibody are covalently linked with a peptide linker to create a bivalent domain antibody. H The regions can target the same or different antigens.

[0065] A "bivalent antibody" contains two antigen-binding sites. In some cases, the two binding sites have the same antigen specificity. However, a bivalent antibody may be bispecific (see below).

[0066] In certain embodiments, the monoclonal antibodies herein also include camelized single domain antibodies. See, e.g., Muyldermans et al. (2001) Trends See Biochem. Sci. 26:230; Reichmann et al. (1999) J. Immunol. Methods 231:25; WO 94 / 04678; WO 94 / 25591; U.S. Patent No. 6,005,079. In one embodiment, the present invention provides a method for the preparation of single domain antibodies comprising two V H A single domain antibody comprising the domain is provided.

[0067] As used herein, the term "diabody" refers to small antibody fragments with two antigen-binding sites, which are composed of a light chain variable domain (V) within the same polypeptide chain. L ) connected to the heavy chain variable domain (V H )(V H -V L , or V L -V H ). By using a linker that is too short to allow pairing between the two domains on the same chain, the domains can be paired with the complementary domains of another chain and create two antigen-binding sites. Diabodies are described more fully in, for example, EP 404,097; WO 93 / 11161; and Holliger et al. (1993) Proc. Natl. Acad. Sci. USA 90: 6444-6448. For a review of recombinant antibody variants, see Holliger and Hudson (2005) Nat. Biotechnol. 23:1126-1136.

[0068] Typically, a variant antibody or antigen-binding fragment of an antibody provided herein retains at least 10% of its IGF-1R binding activity (compared to the modified parent antibody), when activity is expressed on a molar basis. In some embodiments, a variant antibody (or antigen-binding fragment thereof) or antigen-binding fragment of an antibody provided herein retains at least 20%, 50%, 70%, 80%, 90%, 95%, or 100% or more of the IGF-1R binding affinity of the parent antibody. As described herein, it is also intended that the antibodies or antigen-binding fragments of the present invention can include conservative or non-conservative amino acid substitutions, which may also be referred to as "conservative variants" or "function-conservative variants" of the antibody, that do not substantially alter its biological activity.

[0069] "Isolated antibody" refers to a binding compound in a purified state, and in this context means that the molecule is substantially free of other biological molecules, such as nucleic acids, proteins, lipids, carbohydrates, or other materials (such as cellular debris and growth medium). In general, the term "isolated" is not intended to imply the complete absence of such materials, or the absence of water, buffers, or salts, unless such materials are present in amounts that would substantially interfere with the experimental or therapeutic uses of the binding compounds described herein.

[0070] As used herein, the term "monoclonal antibody" refers to a population of substantially homogeneous antibodies, i.e., the antibody molecules comprising the population are identical in amino acid sequence except for possible naturally occurring mutations that may be present in minor amounts. In contrast, conventional (polyclonal) antibody preparations typically include a large number of different antibodies, often with different amino acid sequences in the variable domains (especially the CDRs), specific for different epitopes. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies. However, the term "monoclonal antibody" should not be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies for use in accordance with the present invention may be made by the hybridoma method first described by Kohler et al. (1975) Nature 256:495, or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). The term "monoclonal antibody" also refers to any antibody that is the product of a method described, for example, in Clackson et al., Nature 352:624-628 (1991) and Marks (1991) J. Mol. Biol. 222:581-597. See, e.g., Presta (2005) J. Allergy Clin. Immunol. 116:731.

[0071] As used herein, a "chimeric antibody" is an antibody having a variable domain derived from a first antibody and a constant domain derived from a second antibody, wherein the first and second antibodies are from different species (U.S. Pat. No. 4,816,567 and Morrison et al., (1984) Proc. Natl. Acad. Sci. USA 81: 6851-6855). Typically, the variable domains are obtained from an antibody derived from a laboratory animal such as a rodent (the "parent antibody"), and the constant domain sequences are obtained from a human antibody; therefore, the resulting chimeric antibody is less likely to provoke an adverse immune response in a human subject than the parent (e.g., rodent) antibody.

[0072] As used herein, the term "humanized antibody" refers to forms of antibodies that contain sequences derived from both human and non-human (e.g., mouse, rat) antibodies. Generally, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, with all or substantially all of the hypervariable loops corresponding to those of a non-human immunoglobulin and all or substantially all of the framework (FR) regions being those of a human immunoglobulin sequence. Optionally, the humanized antibody may also comprise at least a portion of a human immunoglobulin constant region (Fc).

[0073] The term "fully human antibody" refers to an antibody that comprises only human immunoglobulin protein sequences. A fully human antibody may contain mouse glycosylation if made in a mouse, a mouse cell, or a hybridoma derived from a mouse cell. Similarly, a "mouse antibody" refers to an antibody that comprises only mouse immunoglobulin sequences. Alternatively, a fully human antibody may contain rat glycosylation if made in a rat, a rat cell, or a hybridoma derived from a rat cell. Similarly, a "rat antibody" refers to an antibody that comprises only rat immunoglobulin sequences.

[0074] Generally, the basic antibody structural unit comprises a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair containing one "light chain" (approximately 25 kDa) and one "heavy chain" (approximately 50-70 kDa). The amino-terminal portion of each chain contains a variable region of approximately 100-110 amino acids in length, primarily responsible for antigen recognition. The carboxy-terminal portion of the heavy chain may define a constant region primarily responsible for effector function. Human light chains are typically classified as kappa and lambda light chains. Human heavy chains are further typically classified as mu, delta, gamma, alpha, or epsilon, defining the antibody isotype as IgM, IgD, IgG, IgA, and IgE, respectively. Within the light and heavy chains, the variable and constant regions are connected by a "J" region of approximately 12 or more amino acids, and the heavy chain also contains a "D" region of approximately 10 or more amino acids. See generally Fundamental Immunology Ch. 7 (Paul, W., ed., 2nd ed. Raven Press, NY (1989)).

[0075] The variable regions of each light / heavy chain pair form the antibody binding site. Thus, an intact antibody generally has two binding sites. Except for bifunctional or bispecific antibodies, two binding sites are The binding sites are generally the same.

[0076] Typically, both heavy and light chain variable domains contain three hypervariable regions, also called complementarity-determining regions (CDRs), located within relatively conserved framework regions (FRs). The CDRs are usually aligned by the framework regions, enabling binding to a specific epitope. Generally, from the N-terminus to the C-terminus, both light and heavy chain variable domains contain FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The amino acid assignments for each domain are generally found in Sequences of Proteins of Immunological Interest, Kabat, et al.; National Institutes of Health, Bethesda, Md.; 5 th ed.;NIH Publ.No.91-3242(1991);Kabat(1978) Adv.Prot.Chem.32:1-75;Kabat, et al.,(1977) J.Biol.Chem.252:6609-6616;Chothia,et al.,(1987) J Mol.Biol.196:901-917 or As defined by Chothia, et al., (1989) Nature 342:878-883.

[0077] As used herein, the term "hypervariable region" refers to the amino acid residues of an antibody that are involved in antigen binding. Hypervariable regions include amino acid residues from the "complementarity determining regions," or "CDRs," (i.e., residues 24-34 (CDRL1), 50-56 (CDRL2), and 89-97 (CDRL3) in the light chain variable domain and residues 31-35 (CDRH1), 50-65 (CDRH2), and 95-102 (CDRH3) in the heavy chain variable domain; Kabat et al. (1991) Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md.) and / or those residues from the "hypervariable loops." (i.e., residues 26-32 (CDRL1), 50-52 (CDRL2), and 91-96 (CDRL3) in the light chain variable domain, and 26-32 (CDRH1), 53-55 (CDRH2), and 96-101 (CDRH3) in the heavy chain variable domain; Chothia and Lesk (1987) J. Mol. Biol. 196: 901-917). As used herein, the term "framework" or "FR" refers to variable domain residues other than the hypervariable region residues defined herein as CDR residues. CDRs provide the majority of contact residues for binding of an antibody to an antigen or epitope. CDRs of interest can be derived from donor antibody variable heavy and light chain sequences and also include analogs of naturally occurring CDRs that share or retain the same antigen-binding specificity and / or neutralizing capacity as the donor antibody from which they are derived.

[0078] The antibodies may be any of the antibodies disclosed in any of the following U.S. patents or patent publications: US7,417,130, US2004 / 132094, US5,831,012, US2004 / 023334, US7,250,297, US6,818,418, US2004 / 209243, US7,838,629, US7,186,524, US6,004,746, US5,475,096, US2004 / 146938, US2004 / 157209, US6,994,982, US6,794,144, US2010 / 239633, US7,803,907, US2010 / 1 19446, and / or US 7,166,697 (the entire contents of each of which are incorporated herein by reference), full-length antibodies, single domain antibodies, recombinant heavy chain only antibodies (VHH), single chain antibodies (scFv), shark heavy chain only antibodies (VNAR), microproteins (cysteine ​​knot proteins, knottins), DARPins; tetranectins; affibodies; transbodies; anticalins; adnectins; affilins; microbodies; peptide aptamers; alterases; plastic antibodies; The antibody may take the form of a phylomer, stradobody, maxibody, shrimpbody, fynomer, armadillo repeat protein, Kunitz domain, avimer, atrimer, probody, immunobody, triomab, troibody, pepbody, vaccibody, unibody, affimer, duobody, Fv, Fab, Fab', F(ab')2, peptidomimetic molecule, or synthetic molecule. See also Storz MAbs. 2011 May-Jun;3(3):310-317, incorporated herein by reference.

[0079] The term "antigen," as used herein, refers to any molecule capable of generating an antibody, or binding to an antibody, either directly or indirectly. The definition of "antigen" includes protein-encoding nucleic acids. "Antigen" can also refer to the binding partner of an antibody. In some embodiments, the antigen is an IGF-1R protein expressed on the cell surface. In some embodiments, the cell is an intact cell. An intact cell is a cell that has not been lysed or disrupted open with a detergent or other reagent. A cell that has been treated with a detergent or other reagent that disrupts or creates holes in the cell membrane is not an intact cell. For example, provided herein is a method for producing an antibody that binds to an IGF-1R protein, the method comprising culturing cells containing a nucleic acid molecule encoding the IGF-1R antibody.

[0080] As used herein, "specific binding," or "immunospecific binding," or "immunospecifically binds" refers to an antibody that binds to a predetermined antigen (e.g., IGF-1R) or epitope present in an antigen. In some embodiments, the antibody binds to a predetermined antigen (e.g., IGF-1R) or epitope present in an antigen. -7 The following dissociation constants (K D ) and binds to nonspecific antigens other than the designated antigen (e.g., BSA, casein, or other nonspecific polypeptides). D At least two times smaller than Dand binds to a predetermined antigen. The phrases "antibody recognizing IGF-1R" and "antibody specific for IGF-1R" are used interchangeably herein with the term "antibody that immunospecifically binds to IGF-1R." In the present disclosure, reference may be made to IGF-1R. The degree of specificity required for an anti-IGF-1R antibody may depend on the intended use of the antibody and, in any event, is defined by its suitability for use for the intended use. In some embodiments, an antibody or binding compound derived from the antigen-binding site of an antibody of the contemplated methods binds to its antigen (IGF-1R) with an affinity that is at least 2-fold greater, at least 10-fold greater, at least 20-fold greater, or at least 100-fold greater than its affinity for any other antigen.

[0081] Methods for measuring mAb specificity and affinity by competitive inhibition are described in Harlow, et al., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1988), Colligan et al., eds., Current Protocols in Immunology, Greene Publishing Assoc. and Wiley Interscience, NY, (1992, 1993), and Muller, Meth. Enzymol. 92:589 601 (1983), which references are incorporated herein by reference in their entireties.

[0082] The term "homolog" refers to a protein sequence having 40% to 100% sequence homology or identity to a reference sequence. The percent identity between two peptide chains can be measured by pairwise alignment using the AlignX module of Vector NTI v.9.0.0 (Invitrogen Corp., Carlsbad, Calif.) with default settings. In some embodiments, the antibody, or antigen-binding fragment thereof, has at least 50, 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence homology or identity to a sequence described herein. or have identity. In some embodiments, the antibodies have conservative substitutions compared to the sequences described herein. Exemplary conservative substitutions are shown in Table 1 and are within the scope of the disclosed subject matter. Conservative substitutions can be made in the framework regions or antigen-binding site, as long as they do not adversely affect the properties of the antibody. Substitutions can be made to improve antibody properties, such as stability or affinity. Conservative substitutions produce molecules with similar functional and chemical properties to the molecule to which such modifications are made. Exemplary amino acid substitutions are shown in the table below. [Table 1]

[0083] In some embodiments, variants of the proteins and peptides provided herein are provided. In some embodiments, the variants comprise substitutions, deletions, or insertions. In some embodiments, the variants comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 (e.g., 1-10) substitutions. The substitutions described herein can be conservative substitutions. In some embodiments, the substitutions are non-conservative. In some embodiments, the variants comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 (e.g., 1-10) deletions. In some embodiments, the variants comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 (e.g., 1-10) insertions. In some embodiments, the substitutions, deletions, or insertions are present in the CDRs provided herein. In some embodiments, the substitutions, deletions, or insertions are not present in the CDRs provided herein.

[0084] The term "in combination" as used herein refers to the use of a mixture of the described agents. This means that they can be administered to an animal or subject simultaneously as a single agent, or sequentially in any order as a single agent.

[0085] Techniques for enlarging antibodies into small peptide sequences that recognize and bind to these sequences when presented in free or conjugated form, or as native sequences in the context of larger proteins, are well known in the art. Such antibodies include mouse, mouse-human, and human-human antibodies produced by hybridoma or recombinant techniques known in the art. Antibodies can also be produced in humans, mice, sheep, rats, rabbits, sharks, llamas, or chickens. In some embodiments, antibodies are produced in chickens. Antibodies can also be produced in other small animals.

[0086] The term "epitope" is meant to refer to a portion of any molecule that can be recognized and bound by an antibody at one or more of the antigen-binding regions of the Ab. Epitopes usually consist of chemically active surface groupings of molecules, such as amino acids or sugar side chains, and have specific three-dimensional structural characteristics as well as specific charge characteristics. Examples of epitopes include, but are not limited to, the residues described herein that form the IGF-1R epitope. In some embodiments, the epitope is present only in the non-denatured protein. In some embodiments, the epitope is present only in the denatured protein.

[0087] In some embodiments, sources of DNA encoding non-human antibodies include cell lines that produce antibodies, such as hybrid cell lines commonly known as hybridomas.

[0088] Hybrid cells are formed by fusing non-human antibody-producing cells, usually spleen cells of animals immunized against either natural or recombinant antigens, or peptide fragments of the antigen protein sequence. Alternatively, the non-human antibody-producing cells can be B lymphocytes obtained from the blood, spleen, lymph nodes, or other tissues of animals immunized with the antigen.

[0089] The second fusion partner, which confers immortalization function, can be a lymphoblastoid cell, a plasmacytoma, or a myeloma cell that is not itself an antibody-producing cell but is a malignant tumor. Fusion partner cells include, but are not limited to, the hybridoma SP2 / 0-Ag14, abbreviated as SP2 / 0 (ATCC CRL1581), and the myeloma P3X63Ag8 (ATCC TIB9), or derivatives thereof. See, e.g., Ausubel, supra; Harlow, supra; and Colligan, supra, the entire contents of which are incorporated herein by reference.

[0090] Antibodies can be generated according to the examples provided herein. Once the sequence is known, the antibody can be generated according to known methods. The antibody can also be converted to a different type, such as a human IgG. By converting the antibody to a human antibody, a human subject should not recognize the antibody as foreign. Converting non-human IgG antibodies to human IgG antibodies is well known and can be done routinely once the native sequence is known. As discussed herein, antibodies can be modified according to known methods. Such methods are described, for example, in Riechmann L, Clark M, Waldmann H, Winter G (1988). "Reshaping human antibodies for therapy." Nature 332(6162): 332-323; Tsurushita N, Park M, Pakabunto K, Ong K, Avdalovic A, Fu H, Jia A, Vasquez M, Kumar S. (2004). Antibody-producing cells involved in the nucleotide sequence encoding the antigen-binding region of the chimeric antibody can be in the form of non-human, e.g., primates, or human cells. Immortal antibody-producing cells can also be produced by transformation. For example, antibody-producing B lymphocytes can be infected with and transformed by a virus such as Epstein-Barr virus, resulting in immortal antibody-producing cells (Kozbor et al., Immunol. Today 4:72-79 (1983)). Alternatively, B lymphocytes can be transformed by providing a transforming gene or a transforming gene product, as is well known. See, e.g., Ausubel, supra, Harlow, supra, and Colligan, supra, the entire contents of which are incorporated herein by reference. Cell fusion is achieved by standard procedures well known to those skilled in the art of immunology. Fusion partner cell lines, as well as methods for fusing and selecting hybridomas and screening for mAbs, are well known in the art. See, e.g., Ausubel, supra, Harlow, supra, and Colligan, supra, the entire contents of which are incorporated herein by reference.

[0091] In some embodiments, the antibody is a MAb that binds to IGF-1R. In some embodiments, the antibody binds to amino acids of an epitope of IGF-1R.

[0092] In some embodiments, the antibody comprises a sequence provided herein.

[0093] The antibody sequences can be modified to obtain human IgG antibodies. The sequences provided herein can be modified to obtain other types of antibodies. The CDRs can also be attached to other antibodies, proteins, or molecules to generate antibody fragments that bind to IGF-1R. This can be present in the form of antibody drug conjugates ("ADCs"), multispecific molecules, or chimeric antigen receptors. The CDRs and antibody sequences provided herein can be humanized or fully human according to known methods. The sequences can also be made into chimeric antibodies as described herein.

[0094] In some embodiments, the antibody comprises an amino acid sequence comprising a sequence provided herein, or a fragment thereof. In some embodiments, the antibody comprises one or more amino acid sequences provided herein, an antigen-binding fragment thereof, or a human IgG variant thereof. By "human IgG variant thereof" is meant an antibody that has been modified to become a human IgG, if the starting antibody is not a human IgG antibody.

[0095] As described herein, production of antibodies using known sequences is routine and can be done by any method. Thus, in some embodiments, nucleic acids encoding antibodies or fragments thereof are provided. In some embodiments, the nucleic acids encode the sequences provided herein. Antibodies can also be modified to become chimeric or human antibodies. Antibodies can also be used in injectable pharmaceutical compositions. Also, as described herein, antibodies can be isolated or recombinant antibodies.

[0096] In some embodiments, "derivatives" of antibodies, fragments, regions, or derivatives thereof are provided (this term includes proteins encoded by truncated or modified genes resulting in molecular species that functionally resemble immunoglobulin fragments). Modifications include, but are not limited to, the addition of genetic sequences encoding cytotoxic proteins such as plant and bacterial toxins. Modifications can also include reporter proteins such as fluorescent or chemiluminescent tags. Fragments and derivatives can be produced by any method.

[0097] Identification of these antigen-binding regions and / or epitopes recognized by the Abs described herein provides the information necessary to generate additional monoclonal antibodies with similar binding characteristics and therapeutic or diagnostic utility comparable to the embodiments of the present application.

[0098] Nucleic acid sequences encoding the antibodies described herein can be genomic DNA or cDNA, or RNA (e.g., mRNA) encoding at least one of the variable regions described herein. A convenient alternative to using chromosomal gene fragments as a source of DNA encoding V region antigen-binding segments is the use of cDNA to construct chimeric immunoglobulin genes, as reported, for example, by Liu et al. (Proc. Natl. Acad. Sci., USA 84:3439 (1987) and J. Immunology 139:3521 (1987)), the entire contents of which are incorporated herein by reference. The use of cDNA requires that the gene be combined with gene expression elements appropriate for the host cell to achieve synthesis of the desired protein. The use of cDNA sequences has an advantage over genomic sequences (containing introns) in that cDNA sequences can be expressed in bacteria or other hosts lacking an appropriate RNA splicing system.

[0099] For example, cDNAs encoding V-region antigen-binding segments capable of detecting, binding to, or neutralizing IGF-1R antigens can be provided using known methods based on the amino acid sequences provided herein. Because the genetic code is degenerate, more than one codon can be used to encode a particular amino acid (Watson, et al., supra). Using the genetic code, two or more different oligonucleotides can be identified, each capable of encoding that amino acid. The probability that a particular oligonucleotide will in fact constitute an actual XXX coding sequence can be estimated by considering unusual base-pairing relationships and the frequency with which particular codons are actually used (and thus encode a particular amino acid) in eukaryotic or prokaryotic cells expressing the antibody or fragment. Such "codon usage rules" are described in Lathe, et al., J. Molec. Biol. 183:1. 12 (1985). The "codon usage rules" of Lathe are used to identify a single oligonucleotide, or a series of oligonucleotides, that contains the theoretically "most probable" nucleotide sequence capable of encoding an antibody variable or constant region sequence.

[0100] The variable regions described herein can be combined with any type of constant region, including human or murine constant regions. Human genes encoding the constant (C) regions of antibodies, fragments, and regions can be obtained from human fetal liver libraries by known methods. Human C region genes can be derived from any human cell, including those that express and produce human immunoglobulins. Human C H The regions can be derived from any of the known classes or isotypes of human heavy chains, including gamma, mu, alpha, gamma, or epsilon, and their subtypes, e.g., G1, G2, G3, and G4. The heavy chain isotypes are responsible for the various effector functions of the antibody and therefore vary in C. HThe choice of region is guided by the desired effector function, such as complement fixation or activity in antibody-dependent cellular cytotoxicity (ADCC). H The regions are derived from γ1 (IgG1), γ3 (IgG3), γ4 (IgG4), or μ (IgM). L The region can be derived from either the kappa or lambda human light chain isotype. In some embodiments, the antibody comprises an Fc region domain. In some embodiments, the Fc domain comprises a mutation that extends the half-life of the antibody. In some embodiments, the Fc domain comprises a mutation such as those described in U.S. Pat. No. 7,670,600, which is incorporated herein by reference in its entirety. In some embodiments, the constant region comprises a mutation at amino acid residue 428, which corresponds to a wild-type human IgG constant domain, numbered according to the EU numbering index of Kabat. Without being bound by any particular theory, antibodies comprising a mutation corresponding to residue 428 may have an increased half-life compared to the half-life of an IgG having a wild-type human IgG constant domain. In some embodiments, the mutation is at a threonine, leucine, The substitution of the native residue with phenylalanine, or serine. In some embodiments, the antibody further comprises one or more amino acid substitutions compared to the corresponding wild-type human IgG constant domain at one or more of amino acid residues 251-256, 285-290, 308-314, 385-389, and 429-436, numbered according to the EU numbering index of Kabat. Specific mutations or substitutions at these positions are described in U.S. Patent No. 7,670,600, which is incorporated by reference in its entirety.

[0101] Genes encoding human immunoglobulin C regions can be obtained from human cells using standard cloning techniques (Sambrook, et al. (Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Press, Cold Spring Harbor, NY (1989) and Ausubel et al., eds. Current Protocols in Molecular Biology (1987-1993)). Human C region genes can be readily obtained from known clones representing two classes of L chains, five classes of H chains, and their subclasses. Chimeric antibody fragments, such as F(ab')2 and Fab, can be prepared by designing appropriately truncated chimeric H-chain genes. For example, a chimeric gene encoding the H-chain portion of an F(ab')2 fragment can include DNA sequences encoding the CH1 domain and hinge region of the H chain, followed by a translation stop codon, resulting in a truncated molecule.

[0102] In some embodiments, the antibodies, murine, human, humanized, or chimeric antibodies, antibody fragments, and regions described herein are prepared by cloning DNA segments encoding the heavy and light chain antigen-binding regions of an IGF-1R antigen-specific antibody, and substituting these DNA segments into the C H and C L These are produced by combining DNA segments encoding these regions to create murine, human, or chimeric immunoglobulin-encoding genes.

[0103] Thus, in some embodiments, fusion chimeric genes are generated that include a first DNA segment encoding at least an antigen-binding region of non-human origin, such as a functionally rearranged V region with a joining (J) segment, joined to a second DNA segment encoding at least a portion of a human C region.

[0104] Thus, a method for producing cDNA encoding antibody V and C regions, an antibody according to some of the embodiments described herein, involves several steps, as exemplified below: 1. isolating and screening messenger RNA (mRNA) from a cell line producing an anti-IGF-1R antigen antibody and from an optional additional antibody that provides the heavy and light constant regions, and producing cDNA therefrom; 2. preparing a full-length cDNA library from the purified mRNA, in which appropriate V and / or C region gene fragments of the L and H chain genes can be (i) identified with appropriate probes, (ii) sequenced, and (iii) matched with C or V gene segments from another antibody to form a chimeric antibody; 3. constructing a complete H or L chain coding sequence by combining cloned specific V region gene fragments with the cloned C region genes described above; and 4. expressing and producing the L and H chains in a selected host, including prokaryotic and eukaryotic cells, to provide a mouse-mouse, human-mouse, human-human, or human-mouse antibody.

[0105] Two coding DNA sequences are said to be "operably linked" when the linkage results in a contiguous, translatable sequence without alteration or interruption of the triplet reading frame. A DNA coding sequence is operably linked to a gene expression element when the linkage results in the proper function of the gene expression element and expression of the coding sequence.

[0106] As used herein, and unless otherwise specified, the term "about" is intended to mean ±5% of the value it modifies. Thus, about 100 means 95 to 105.

[0107] In some embodiments, the antibodies described herein are used to detect the presence of an antigen. The antibodies can be used in any device or method to detect the presence of the antigen.

[0108] The term "purified" when referring to an antibody means an antibody that is substantially free of other materials that are associated with the molecule in its natural environment. For example, a purified protein is substantially free of cellular material or other proteins from the cell or tissue from which it is derived. The term refers to preparations where the isolated protein is sufficiently pure for analysis, or is at least 70%-80% (w / w) pure, at least 80%-90% (w / w) pure, 90-95% pure; and at least 95%, 96%, 97%, 98%, 99%, or 100% (w / w) pure. In some embodiments, an antibody is purified.

[0109] As an alternative to preparing monoclonal antibody-secreting hybridomas, monoclonal antibodies against a polypeptide can be identified and isolated by screening a recombinant combinatorial immunoglobulin library (e.g., an antibody phage display library) with the polypeptides described herein to isolate immunoglobulin library members that bind to the polypeptide. Techniques and commercially available kits for generating and screening phage display libraries are well known to those skilled in the art. Furthermore, examples of methods and reagents particularly suitable for use in generating and screening antibody or antigen-binding protein display libraries can be found in the literature. Thus, the epitopes described herein can be used to screen for other antibodies that can be used therapeutically, diagnostically, or as research tools.

[0110] antibody conjugates

[0111] The antibodies provided herein can also be conjugated to a chemical moiety. The chemical moiety can be, among others, a polymer, a radionuclide, or a cytotoxic agent. In some embodiments, this may be referred to as an antibody-drug conjugate. In some embodiments, the chemical moiety is a polymer that increases the antibody molecule's activity in the subject's body. Suitable polymers include, but are not limited to, polyethylene glycol (PEG) (e.g., 2 kDa, 5 kDa, 10 kDa, 12 kDa, 20 kDa, 30 kDa, or 40 kDa), dextran, and monomethoxypolyethylene glycol (mPEG). Lee, et al., (1999) (Bioconj. Chem. 10:973-981) discloses PEG-conjugated single-chain antibodies. Wen, et al. (2001) (Bioconj. Chem. 12:545-553) discloses the conjugation of antibodies with PEG linked to a radiometal chelator (diethylenetriaminepentaacetic acid (DTPA)). Examples of chemical moieties include, but are not limited to, antimitotic agents such as calicheamicins (e.g., ozogamicin), monomethyl auristatin E, mertansine, and the like. Other examples include, but are not limited to, biologically active microtubule inhibitors, alkylating agents, and DNA minor groove binders. Further examples are provided herein and below. Chemical moieties can be attached to antibodies via linking groups (aminobenzyl), cleavable linkers, e.g., cathepsin-cleavable linkers (valine-citrulline), and, in some embodiments, one or more spacers (e.g., p-aminobenzyl carbamate). Without being bound to any particular theory, it is believed that when the antibody conjugate binds to IGF-1R, the antibody conjugate can be internalized and the chemical moiety can kill the cell. In some embodiments, the cell is a thyroid cell.

[0112] The antibodies and antibody fragments of the present invention are 99 Tc, 90 Y,111 In, 32 P, 14 C. 125 I, 3 H, 131 I, 11 C. 15 O. 13 N, 18 F, 35 S, 51 Cr, 57 To, 226 Ra, 60 Co, 59 Fe, 57 Se, 152 EU, 67 CU, 217 Ci, 211 At, 212 Pb, 47 Sc, 109 Pd, 234 Th, and 40 K. 157 Gd, 55 Mn, 52 Tr, and 56 It is also possible to conjugate with a label such as Fe.

[0113] Antibodies and antibody fragments include rare earth chelates, fluorescein and its derivatives, rhodamine and its derivatives, isothiocyanates, phylcoerythrin, phycocyanin, allophycocyanin, o-phthalaldehyde, fluorescamine, 152 Conjugation with fluorescent or chemiluminescent labels is also possible, including fluorophores such as Eu, dansyl, umbelliferone, luciferin, luminal labels, isoluminal labels, aromatic acridinium ester labels, imidazole labels, acridinium salt labels, oxalate ester labels, aequorin labels, 2,3-dihydrophthalazinedione, biotin / avidin, spin labels, and stable free radicals.

[0114] Antibody molecules can also be conjugated to cytotoxic agents such as diphtheria toxin, Pseudomonas aeruginosa exotoxin A chain, ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteins and compounds (e.g., fatty acids), dianthin proteins, Phytoiacca americana proteins PAPI, PAPII, and PAP-S, momordica charantia inhibitor, curcin, crotin, saponaria officinalis inhibitor, mitogenin, restrictocin, phenomycin, and enomycin.

[0115] Any method known in the art can be used to conjugate the antibody molecules of the present invention to various moieties, including those described by Hunter, et al., (1962) Nature 144:945; David, et al., (1974) Biochemistry 13:1014; Pain, et al., (1981) J. Immunol. Meth. 40:219; and Nygren, J., (1982) Histochem. and Cytochem. 30:407. Methods for conjugating antibodies are conventional and very well known in the art.

[0116] Chimeric Antigen Receptor

[0117] The antibodies provided herein can also be incorporated into chimeric antigen receptors ("CARs") that can be used, for example, in CAR-T cells. In some embodiments, the extracellular domain of the CAR can be an antibody provided herein. In some embodiments, the antibody is in scFv format. CAR-T cells are a type of treatment in which a patient's T cells are modified to attack cells that express IGF-1R. The cells are collected from the patient's blood. A special receptor that binds to a specific protein on the patient's cells is then added in the laboratory. In some embodiments, the receptor binds to IGF-1R using the binding region of an antibody provided herein. CAR-T cells containing the IGF-1R antibody can then be used to treat conditions such as those provided herein.

[0118] In some embodiments, the antibody (e.g., an anti-IGF-1R antibody) is In some embodiments, the antibody is a recombinant antibody that binds to IGF-1R protein. In some embodiments, the IGF-1R protein is human IGF-1R protein. In some embodiments, the IGF-1R protein recognized by the antibody is in its native (non-denatured) conformation. In some embodiments, the antibody does not specifically bind to denatured IGF-1R protein. As used herein, the term "recombinant antibody" refers to an antibody that does not occur in nature. In some embodiments, the term "recombinant antibody" refers to an antibody that is not isolated from a human subject.

[0119] In some embodiments, the antibody comprises one or more peptides having the following sequences, or variants thereof: [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7]

[0120] In some embodiments, the antibody comprises one or more peptides having the following sequences, or variants thereof: [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4]

[0121] The columns shown as antibody sequences include the VH and VL chains of the antibody. In cases where a VH chain is shown along with an Fc sequence, the Fc sequence can be modified or replaced with a different Fc region as provided herein. However, in some embodiments, an antibody can comprise the VH and VL sequences provided with respect to the tables provided herein. The antibody comprises one or more VH, HC, LC, or VL (where the sequences with the constant domains are the complete light or heavy chain) having the following sequences, or variants thereof: [Table 4-1] [Table 4-2] [Table 4-3] [Table 4-4] [Table 4-5]

[0122] In some embodiments, the variable light chain set forth as SEQ ID NO: 13 does not have a C-terminal arginine residue, as shown, for example, in the following sequence: DVVMTQTPLSLPVSLGDPASISCRSSQSIVHSNVNTYLEWYLQKPGQSPRLLIYKVSNRFSGVPDRFSGSGAGTDFTLRISRVEAEDLGIYYCFQGSHVPPTFGGGTKLEIK (SEQ ID NO: 97).

[0123] Thus, in some embodiments in which the variable light chain comprises the sequence of SEQ ID NO:13, the sequence of SEQ ID NO:13 can be replaced with the sequence of SEQ ID NO:97.

[0124] In some embodiments, the heavy chain variable region set forth in SEQ ID NO: 14 can comprise a C22S substitution, as shown in the following sequence: QVQLVQSGAEVVKPGASVKLSSKASGYTFTSYWMHWVKQRPGQGLEWIGEINPSNGRTNYNQKFQGKATLTVDKSSSTAYMQLSSLTSEDSAVYYFARGRPDYYGSSKWYFDVWGQGTTVTVSS (SEQ ID NO: 96).

[0125] Thus, in some embodiments, the antibody comprises the VH sequence of SEQ ID NO:96 and the VL sequence of SEQ ID NO:13 or SEQ ID NO:97.

[0126] In some embodiments, the antibody comprises the VH sequence of SEQ ID NO:14 and the VL sequence of SEQ ID NO:97.

[0127] In some embodiments, the antibody comprises a VL of SEQ ID NO: 98 and a VH of SEQ ID NO: 99. In some embodiments, the antibody comprises a VL of SEQ ID NO: 98 and a VH of SEQ ID NO: 99 with an Fc region comprising M252Y, S254T, and T256E mutations. In some embodiments, the antibody comprises a VL of SEQ ID NO: 98 and a VH of SEQ ID NO: 99 with an Fc region comprising M428L and N434S mutations.

[0128] As provided herein, the heavy chain can be linked to an Fc region, including those with mutations that may affect the half-life of the antibody. Non-limiting mutations in the Fc region are provided herein.

[0129] In the tables provided herein, the LC and HC may be shown with the VH and VL domains, with or without the constant region. The constant region may be replaced as provided herein. The VH and VL regions may be used to form the antibodies provided herein. The VH and VL sequences may be present in any format, including, but not limited to, an scFv format in which the VH and VL regions are linked with a peptide linker. Peptides that can be used to bind various peptides provided herein An example of a linker is (GGGGS) n (SEQ ID NO: 73); (GGGGA) n(SEQ ID NO: 74), or any combination thereof, wherein each n is independently 1 to 5. In some embodiments, the variable region is not connected with a peptide linker. In some embodiments, the antibody comprises SEQ ID NO: 1 and SEQ ID NO: 2, or CDR regions thereof. In some embodiments, the antibody comprises SEQ ID NO: 3 and SEQ ID NO: 4, or CDR regions thereof. In some embodiments, the antibody comprises SEQ ID NO: 5 and SEQ ID NO: 6, or CDR regions thereof. In some embodiments, the antibody comprises SEQ ID NO: 7 and SEQ ID NO: 8, or CDR regions thereof. In some embodiments, the antibody comprises SEQ ID NO: 9 and SEQ ID NO: 10, or CDR regions thereof. In some embodiments, the antibody comprises SEQ ID NO: 11 and SEQ ID NO: 12, or CDR regions thereof. In some embodiments, the antibody comprises SEQ ID NO: 13 and SEQ ID NO: 14, or CDR regions thereof. In some embodiments, the antibody comprises SEQ ID NO: 15 and SEQ ID NO: 16, or CDR regions thereof.

[0130] In some embodiments, an antibody or antigen-binding fragment thereof is provided, wherein the antibody or antibody fragment comprises a peptide selected from the table below. [Table 5]

[0131] In some embodiments, the antibody, or antibody-binding fragment thereof, comprises a heavy chain or light chain CDR having the sequence of SEQ ID NO: 17-64, and 81. In some embodiments, the antibody, or antibody-binding fragment thereof, comprises a light chain CDR having the sequence of SEQ ID NO: 17, 18, 19, 23, 24, 25, 29, 30, 31, 35, 36, 37, 41, 42, 43, 47, 48, 49, 53, 54, 55, 59, 60, 61, or 81. In some embodiments, the antibody, or antibody-binding fragment thereof, comprises a heavy chain CDR having the sequence of SEQ ID NO: 20, 21, 22, 26, 27, 28, 32, 33, 34, 38, 39, 40, 44, 45, 46, 50, 51, 52, 56, 57, 58, 62, 63, or 64.

[0132] In some embodiments, the antibody, or antibody-binding fragment thereof, comprises a light chain having LCDR1, LCDR2, and LCDR3, wherein the LCDR1 has the sequence of SEQ ID NO: 17, 23, 29, 35, 41, 47, 53, or 59, the LCDR2 has the sequence of SEQ ID NO: 18, 24, 30, 36, 42, 48, 54, or 60, and the LCDR3 has the sequence of SEQ ID NO: 19, 25, 31, 37, 43, 49, 55, 61, or 81.

[0133] In some embodiments, the antibody, or antibody-binding fragment thereof, comprises a heavy chain having HCDR1, HCDR2, and HCDR3, wherein HCDR1 has the sequence of SEQ ID NO: 20, 26, 32, 38, 44, 50, 56, or 62; HCDR2 has the sequence of SEQ ID NO: 21, 27, 33, 39, 45, 51, 57, or 63; and HCDR3 has the sequence of SEQ ID NO: 22, 28, 34, 40, 46, 52, 58, or 64.

[0134] Different CDR motifs can be combined in any combination, including those not shown in the table above. For example, the following embodiments are provided as non-limiting examples of such combinations:

[0135] In some embodiments, the antibody or antigen-binding fragment thereof comprises: (i) a light chain variable region comprising a light chain CDR1, CDR2, and CDR3 sequence, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 17; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 18; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 19; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2, and CDR3 sequence, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 20; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 21; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 22; or a variant of any of the foregoing.

[0136] In some embodiments, the antibody or antigen-binding fragment thereof comprises: (i) a light chain variable region comprising a light chain CDR1, CDR2, and CDR3 sequence, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 23; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 24; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 25; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2, and CDR3 sequence, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 26; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 27; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 28; or a variant of any of the foregoing.

[0137] In some embodiments, the antibody or antigen-binding fragment thereof comprises: (i) a light chain variable region comprising a light chain CDR1, CDR2, and CDR3 sequence, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 29; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 30; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 31; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2, and CDR3 sequence, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 32; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 33; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 34; or a variant of any of the foregoing.

[0138] In some embodiments, the antibody or antigen-binding fragment thereof comprises: (i) a light chain variable region comprising a light chain CDR1, CDR2, and CDR3 sequence, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 35; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 36; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 37; and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 38; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 39; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 40; or a variant of any of the foregoing.

[0139] In some embodiments, the antibody or antigen-binding fragment thereof comprises: (i) a light chain variable region comprising a light chain CDR1, CDR2, and CDR3 sequence, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 41; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 42; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 43; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2, and CDR3 sequence, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 44; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 45; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 46; or a variant of any of the foregoing.

[0140] In some embodiments, the antibody or antigen-binding fragment thereof comprises: (i) a light chain variable region comprising a light chain CDR1, CDR2, and CDR3 sequence, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 47; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 48; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 49; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2, and CDR3 sequence, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 50; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 51; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 52; or a variant of any of the foregoing.

[0141] In some embodiments, the antibody or antigen-binding fragment thereof comprises: (i) a light chain variable region comprising a light chain CDR1, CDR2, and CDR3 sequence, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 53; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 54; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 55; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2, and CDR3 sequence, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 56; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 57; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 58; or a variant of any of the foregoing.

[0142] In some embodiments, the antibody or antigen-binding fragment thereof comprises: (i) a light chain variable region comprising a light chain CDR1, CDR2, and CDR3 sequence, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 59; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 60; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 61; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2, and CDR3 sequence, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 62; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 63; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 64; or a variant of any of the foregoing.

[0143] In some embodiments, the antibody or antigen-binding fragment thereof comprises: (i) a light chain variable region comprising a light chain CDR1, CDR2, and CDR3 sequence, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 35; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 36; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 37; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2, and CDR3 sequence, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 38; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 39; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 40; or a variant of any of the foregoing.

[0144] In some embodiments, the light chain variable region CDR1 is replaced with any of other light chain CDR1 sequences. In some embodiments, the light chain variable region CDR2 is replaced with any of other light chain CDR2 sequences. In some embodiments, the light chain variable region CDR3 is replaced with any of other light chain CDR3 sequences. In some embodiments, the heavy chain variable region CDR1 is replaced with any of other light chain CDR1 sequences. In some embodiments, the heavy chain variable region CDR2 is replaced with any of other light chain CDR2 sequences. In some embodiments, the heavy chain variable region CDR3 is replaced with any of other light chain CDR3 sequences.

[0145] In some embodiments, an antibody, or antigen-binding fragment thereof, or protein is provided, comprising a peptide having a sequence set forth in any of SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 79, or 86, and 2, 4, 6, 8, 10, 12, 14, 16, 80, or 83.

[0146] In some embodiments, the antibody or antigen-binding fragment thereof comprises any of the foregoing sequences, or any of the foregoing variants.

[0147] In some embodiments, the antibody or antigen-binding fragment thereof comprises the sequence of SEQ ID NO: 65, or any of the foregoing variants. In some embodiments, the antibody or antigen-binding fragment thereof comprises the sequence of SEQ ID NO: 66, or any of the foregoing variants. In some embodiments, the antibody or antigen-binding fragment thereof comprises the sequence of SEQ ID NO: 67, or any of the foregoing variants. In some embodiments, the antibody or antigen-binding fragment thereof comprises the sequence of SEQ ID NO: 68, or any of the foregoing variants. In some embodiments, the antibody or antigen-binding fragment thereof comprises the sequence of SEQ ID NO: 69, or any of the foregoing variants. In some embodiments, the antibody or antigen-binding fragment thereof comprises the sequence of SEQ ID NO: 70, or any of the foregoing variants. In some embodiments, the antibody or antigen-binding fragment thereof comprises the sequence of SEQ ID NO: 71, or any of the foregoing variants. In some embodiments, the antibody or antigen-binding fragment thereof comprises the sequence of SEQ ID NO: 72, or any of the foregoing variants. In some embodiments, the antibody or antigen-binding fragment thereof comprises the sequence of SEQ ID NO: 78, or any of the foregoing variants. In some embodiments, the antibody or antigen-binding fragment thereof comprises the sequence of SEQ ID NO: 82, or any of the foregoing variants. In some embodiments, the antibody or antigen-binding fragment thereof comprises the sequence of SEQ ID NO: 85, or a variant of any of the foregoing.

[0148] In some embodiments, V L and / or V H The sequences are as described herein. In some embodiments, V L The sequences are provided as elements of the light chain (LC). In some embodiments, the V sequences are provided as elements of the light chain (LC). L The V sequences are underlined as LC sequences. In some embodiments, the V sequences are provided as elements of the heavy chain (LC). H The sequence is underlined in the HC sequence.

[0149] In some embodiments, the antibody or antigen-binding fragment thereof comprises a V sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 79, or 86. L peptides, or any combination thereof. L The peptides can include variants of any of these sequences provided herein.

[0150] In some embodiments, the antibody or antigen-binding fragment thereof comprises a V sequence set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 80, or 83. H peptides, or any combination thereof. H The peptides can include variants of any of these sequences provided herein.

[0151] In some embodiments, the antibody or antigen-binding fragment thereof is H Peptides and V L A peptide, H The peptide comprises a sequence set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 80, or 83, L The above V, wherein the peptide comprises a sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 79, or 86. H Peptides and V L Contains peptides.

[0152] In some embodiments, the antibody or antigen-binding fragment thereof is H Peptides and V L A peptide, H The peptide comprises the sequence set forth in SEQ ID NO: 2, L The peptide comprises the sequence set forth in SEQ ID NO: 1. H Peptides and V L In some embodiments, the antibody or antigen-binding fragment thereof comprises a V H Peptides and V L A peptide, H The peptide comprises the sequence set forth in SEQ ID NO: 4,L The peptide comprises the sequence set forth in SEQ ID NO: 3. H Peptides and V L In some embodiments, the antibody or antigen-binding fragment thereof comprises a V H Peptides and V L A peptide, H The peptide comprises the sequence set forth in SEQ ID NO:6, L The peptide comprises the sequence set forth in SEQ ID NO: 5. H Peptides and V L In some embodiments, the antibody or antigen-binding fragment thereof comprises a V H Peptides and V L A peptide, H The peptide comprises the sequence set forth in SEQ ID NO: 8, L The peptide comprises the sequence set forth in SEQ ID NO: 7. H Peptides and V L In some embodiments, the antibody or antigen-binding fragment thereof comprises a V H Peptides and V L A peptide, H The peptide comprises the sequence set forth in SEQ ID NO: 10, L The peptide comprises the sequence set forth in SEQ ID NO: 9. H Peptides and V L In some embodiments, the antibody or antigen-binding fragment thereof comprises a V H Peptides and V L A peptide, H The peptide comprises the sequence set forth in SEQ ID NO: 12, L The peptide comprises the sequence set forth in SEQ ID NO: 11. H Peptides and V L In some embodiments, the antibody or antigen-binding fragment thereof comprises a V H Peptides and V L A peptide, H The peptide comprises the sequence set forth in SEQ ID NO: 14, LThe peptide comprises the sequence set forth in SEQ ID NO: 13. H Peptides and V L In some embodiments, the antibody or antigen-binding fragment thereof comprises a V H Peptides and V L A peptide, H The peptide comprises the sequence set forth in SEQ ID NO: 16, L The peptide comprises the sequence set forth in SEQ ID NO: 15. H Peptides and V L In some embodiments, the antibody or antigen-binding fragment thereof comprises a V H Peptides and V L A peptide, H The peptide comprises the sequence set forth in SEQ ID NO: 80, L The peptide comprises the sequence set forth in SEQ ID NO: 79. H Peptides and V L In some embodiments, the antibody or antigen-binding fragment thereof comprises a V H Peptides and V L A peptide, H The peptide comprises the sequence set forth in SEQ ID NO: 83, L The peptide comprises the sequence set forth in SEQ ID NO: 3. H Peptides and V L In some embodiments, the antibody or antigen-binding fragment thereof comprises a V H Peptides and V L A peptide, H The peptide comprises the sequence set forth in SEQ ID NO: 14, L The peptide comprises the sequence set forth in SEQ ID NO: 86. H Peptides and V L Contains peptides.

[0153] In some embodiments, the antibody or antigen-binding fragment thereof comprises an LC peptide set forth in SEQ ID NO: 1, 3, 5, 7, 9, or 11, or any combination thereof. The LC peptide can include variants of any of these sequences provided herein.

[0154] In some embodiments, the antibody or antigen-binding fragment thereof comprises an HC peptide set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, or 83, or any combination thereof. The HC peptide can include variants of any of these sequences provided herein.

[0155] In some embodiments, the antibody or antigen-binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises the sequence set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, or 83, and the LC peptide comprises the sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, or 11. In some embodiments, the antibody or antigen-binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises the sequence set forth in SEQ ID NO: 2, and the LC peptide comprises the sequence set forth in SEQ ID NO: 1. In some embodiments, the antibody or antigen-binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises the sequence set forth in SEQ ID NO: 4, and the LC peptide comprises the sequence set forth in SEQ ID NO: 3. In some embodiments, the HC peptide comprising the sequence set forth in SEQ ID NO: 4 has an additional C-terminal lysine (K) residue. In some embodiments, the antibody or antigen-binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises the sequence set forth in SEQ ID NO: 6 and the LC peptide comprises the sequence set forth in SEQ ID NO: 5. In some embodiments, the antibody or antigen-binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises the sequence set forth in SEQ ID NO: 8 and the LC peptide comprises the sequence set forth in SEQ ID NO: 7. In some embodiments, the antibody or antigen-binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises the sequence set forth in SEQ ID NO: 10 and the LC peptide comprises the sequence set forth in SEQ ID NO: 9. In some embodiments, the antibody or antigen-binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises the sequence set forth in SEQ ID NO: 12 and the LC peptide comprises the sequence set forth in SEQ ID NO: 11.In some embodiments, the antibody or antigen-binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises the sequence set forth in SEQ ID NO: 83 and the LC peptide comprises the sequence set forth in SEQ ID NO: 3.

[0156] In addition to these specific combinations, V H Peptides and V L Any of the peptides can be combined with each other.

[0157] In addition to these specific combinations, any of the HC and LC peptides can be combined with each other.

[0158] In some embodiments, the antibody comprises the sequence of ATCC clone PTA-7444, or an antigen-binding fragment thereof. The sequence of the antibody produced by ATCC clone PTA-7444 is incorporated herein by reference, including antigen-binding fragments thereof.

[0159] Additionally, as provided herein, antibodies can be multispecific, in that they have multiple binding regions that target different proteins or the same protein at different epitopes. In some embodiments, the antibodies are bispecific.

[0160] As provided herein, the different peptides described herein (V H or V L ) can be attached to a peptide linker, or can be attached to a contiguous sequence without being attached to a peptide linker. In some embodiments, the peptide linker is (GGGGS) n (SEQ ID NO: 73); (GGGGA) n (SEQ ID NO: 74), or any combination thereof, wherein each n is independently 1 to 5. The conjugated peptide format includes the sequence of V H -ZV L or V L -ZV Hwherein Z is a peptide linker In some embodiments, Z can be represented by the formula: (GGGGS) n (SEQ ID NO: 73); (GGGGA) n (SEQ ID NO: 74), or any combination thereof (wherein each n is independently 1 to 5).

[0161] As provided herein, antibodies or antigen-binding fragments thereof can be sequence variants.

[0162] Other exemplary antibodies include US20160096894A1, EP1399483B1, EP2194067B1, US20040202651A1, US20110229933A1, US8137933B2, US8951790B2, US20190270820A1, US7572897B2, US20090275126A1, EP1959014B1, US20080014203A1, US20080226635A1, US20120076778A1, US20190153071A1, WO2011161119A1, and US1 0611825B2, US20120237507A1, EP2681240B1, US9982036B2, US20180312573A1, EP2681239B1, US20160151487A1, US20190225696A1, WO2017011773A2, US20200023076A1, US20190153471A1, US20190194713A1, WO2020006486A1, US20080112888A1, US20150168424A1, EP2032989B2, US9045536B2.Other exemplary antibodies include those described in US8153121B2, EP1469879B1, WO2016064716A1, US20190270820A1, US20180280527A1, US20190225696A1, US7998681B2, US20040202651A1, US2005 0136063A1, US20090285824A1, US20150274829A1, EP2322550B1, US20060286103A1, US2007007 1675A1, US20100047239A1, US20130004416A1, US20080112888A1, US20150168424A1, US201001 43340A1, US20110014117A1, US20100260668A1, US20100074900A1, US20150017168A1, US20110 044980A1, US20130330323A1, US20120263722A1, US20120201746A1, US10519245B2, US2018024 3432A1, US20170218091A1, US20200115460A1, US20100104645A1, US20120065380A1, EP2970433B1, US20160289341A1, US20160289343A1, US20190293656A1.

[0163] In some embodiments, the antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises: QVQLVQSGAEVVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGEINPSNGRTNYNQKFQGKATLTVDKSSSTAYMQLSSLTSEDSAVYYFARGRPDYYGSSKWY FDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK KVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPGK (SEQ ID NO: 92), and the light chain comprises DVVMTQTPLSLPVSLGDPASISCRSSQSIVHSNVNTYLEWYLQKPGQSPRLLIYKVSNRFSGVPDRFSGSGAGTDFTLRISRVEAEDLGIYYCFQGSHVPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 93).

[0164] In some embodiments, the antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises: QVQLVQSGAEVVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGEINPSNGRTNYNQKFQGKATLTVDKSSSTAYMQLSSLTSEDSAVYYFARGRPDYYGSSKWYFDVWGQGT TVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP APELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 94), DVVMTQTPLSLPVSLGDPASISCRSSQSIVHSNVNTYLEWYLQKPGQSPRLLIYKVSNRFSGVPDRFSGSGAGTDFTLRISRVEAEDLGIYYCFQGSHVPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 93).

[0165] In some embodiments, the heavy chain of SEQ ID NO:94 comprises a C-terminal lysine residue added to the C-terminus of SEQ ID NO:94.

[0166] In some embodiments, the antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises: QVQLVQSGAEVVKPGASVKLSSKASGYTFTSYWMHWVKQRPGQGLEWIGEINPSNGRTNYNQKFQGKATLTVDKSSSTAYMQLSSLTSEDSAVYYFARGRPDYYGSSKWYFDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFL FPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 95), and the light chain comprises the sequence of SEQ ID NO: 93.

[0167] In some embodiments, the heavy chain of SEQ ID NO:95 comprises a C-terminal lysine residue added to the C-terminus of SEQ ID NO:95.

[0168] In some embodiments, the antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the sequence of SEQ ID NO:83 and the light chain comprises the sequence of SEQ ID NO:3.

[0169] In some embodiments, the antibody comprises the VH sequence of SEQ ID NO: 96 and the VL sequence of SEQ ID NO: 13 or SEQ ID NO: 97. In some embodiments, the antibody comprises the VH sequence of SEQ ID NO: 14 and the VL sequence of SEQ ID NO: 97.

[0170] Pharmaceutical Composition

[0171] In some embodiments, to prepare pharmaceutical or sterile compositions of anti-IGF-1R antibodies, or other proteins provided herein, the antibodies or antigen-binding fragments thereof, or other proteins provided herein, are mixed with a pharmaceutically acceptable carrier or excipient. See, e.g., Remington's Pharmaceutical Sciences and US Pharmacopeia: National Formulary, Mack Publishing Company, Easton, PA (1984).

[0172] Formulations of therapeutic and diagnostic agents can be prepared, for example, by mixing with an acceptable carrier, excipient, or stabilizer in the form of a lyophilized powder, a slurry, an aqueous solution, or a suspension (see, e.g., Hardman, et al. (2001) Goodman and Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill, New York, NY; Gennaro (2000) Remington: The Science and Practice of Pharmacy, Lippincott, Williams, and Wilkins, New York, NY; Avis, et al. (eds.) (1993) Pharmaceutical Dosage Forms: Parenteral Medications, Marcel Dekker, NY; Lieberman, et al. (eds.) (1990) Pharmaceutical Dosage Forms: Tablets, Marcel Dekker, NY; Lieberman, et al. (eds.) (1990) Pharmaceutical Dosage Forms: Disperse Systems, Marcel (See Dekker, NY; Weiner and Kotkoskie (2000) Excipient Toxicity and Safety, Marcel Dekker, Inc., New York, NY). In some embodiments, the antibody is diluted to the appropriate concentration in sodium acetate solution (pH 5-6), with NaCl or sucrose added for tonicity. Additional agents, such as polysorbate 20 or polysorbate 80, can be added to improve stability.

[0173] The toxicity and therapeutic efficacy of an antibody composition administered alone or in combination with another agent may be measured, for example, by the LD 50 (the dose lethal to 50% of the population), and ED 50The dose that is therapeutically effective in 50% of a population can be determined by standard pharmaceutical procedures in cell culture or experimental animals. The dose ratio between toxic and therapeutic effects is called the therapeutic index (LD 50 / ED 50 ) In certain embodiments, antibodies that exhibit large therapeutic indices are desirable. The data obtained from these cell culture assays and animal studies can be used to formulate a range of dosages for use in humans. The dosage of such compounds is preferably within the ED 50 The dose may vary within this range depending on the dosage form used and the route of administration.

[0174] In some embodiments, the compositions of the present invention are administered according to the Physician's Guide 2003 (Thomson Pharmacy). Subjects will be administered the test in accordance with the Guidelines for Healthcare; 57th edition (November 1, 2002).

[0175] The method of administration can be varied: Suitable routes of administration include oral, rectal, transmucosal, intestinal, parenteral, intramuscular, subcutaneous, intradermal, intramedullary, intrathecal, direct intraventricular, intravenous, intraperitoneal, nasal, intraocular, aspiration, inhalation, topical, cutaneous, transdermal, or intraarterial.

[0176] In some embodiments, the antibody or antigen-binding fragment thereof can be administered by an invasive route, such as injection. In some embodiments, the antibody or antigen-binding fragment thereof, or pharmaceutical composition thereof is administered intravenously, subcutaneously, intramuscularly, intraarterially, intraarticularly (e.g., into an arthritic joint), or by inhalation or aerosol delivery. Administration by a non-invasive route (e.g., orally, e.g., in a pill, capsule, or tablet) is also within the scope of this embodiment.

[0177] In some embodiments, the antibody or antigen-binding fragment thereof can be administered directly to the eye, the anterior chamber of the eye, the vitreous chamber of the eye, the suprachoroidal space, or the retroorbital space. In some embodiments, administration to the eye, the anterior chamber of the eye, the vitreous chamber of the eye, the suprachoroidal space, or the retroorbital space is by injection. In some embodiments, the injection is an intravitreal injection, an intraorbital injection, a retroorbital injection, a suprachoroidal injection, or an intracavitary injection. In some embodiments, the injection is an intravitreal injection. In some embodiments, the injection is an intraorbital injection. In some embodiments, the injection is a retroorbital injection. In some embodiments, the injection is a suprachoroidal injection. In some embodiments, the injection is an intracavitary injection.

[0178] In some embodiments, the anti-IGF-1R antibody, or antigen-binding fragment thereof, is administered in combination with at least one additional therapeutic agent, such as, but not limited to, any therapeutic agent used to treat thyroid eye disease. For example, in some embodiments, the anti-IGF-1R antibody, or antigen-binding fragment thereof, is administered in combination with at least one additional therapeutic agent, such as, but not limited to, a therapeutic agent used to treat thyroid eye disease or a condition associated with thyroid eye disease. Examples of such therapeutic agents and agents include antithyroid medications, diabetes medications, beta-blockers, propylthiouracil, methimazole, propranolol, atenolol, metoprolol, nadolol, corticosteroids, metformin, sulfonylureas, meglitinides, thiazolidinediones, DPP-4 inhibitors, GLP-1 receptor agonists, SGLT2 inhibitors, regular insulin, insulin aspart, insulin glulisine, insulin lispro, insulin isophane, insulin degludec, insulin detemir, insulin glargine, acarbose, miglitol, and acarbose. These include, but are not limited to, sebutolol, atenolol, betaxolol, bisoprolol, carteolol, carvedilol, esmolol, labetalol, metoprolol, nadolol, nebivolol, penbutolol, pindolol, propranolol, sotalol, timolol, tomolol, ophthalmic solution, sitagliptin, saxagliptin, linagliptin, alogliptin, dulaglutide, exenatide, semaglutide, liraglutide, lixisenatide, canagliflozin, dapagliflozin, empagliflozin, or any combination thereof.

[0179] Compositions can be administered using medical devices known in the art, for example, pharmaceutical compositions of the invention can be administered by injection using a hypodermic needle, including, for example, a prefilled syringe or an auto-injector.

[0180] The pharmaceutical compositions may be prepared using methods described in U.S. Patent Nos. 6,620,135; 6,096,002; 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880; 4,790,824, or Administration can also be accomplished using a needleless hypodermic injection device, such as the device disclosed in US Pat. No. 4,596,556.

[0181] Pharmaceutical compositions can also be administered by infusion. Examples of well-known implant and module forms for administering pharmaceutical compositions include U.S. Pat. No. 4,487,603, which discloses an implantable microinfusion pump for dispensing drugs at a controlled rate; U.S. Pat. No. 4,447,233, which discloses a drug infusion pump for delivering drugs at a precise infusion rate; U.S. Pat. No. 4,447,224, which discloses an implantable infusion device with variable flow for continuous drug delivery; and U.S. Pat. No. 4,439,196, which discloses an osmotic drug delivery system with multiple chamber compartments. Many other such implants, delivery systems, and modules are known to those skilled in the art.

[0182] Alternatively, antibodies can be administered locally rather than systemically, for example, by injecting them directly into arthritic joints or pathogen-induced lesions identified by immunopathology, often in a depot or sustained-release formulation. Furthermore, antibodies can be administered in targeted drug delivery systems, for example, in liposomes coated with tissue-specific antibodies that target arthritic joints or pathogen-induced lesions identified by immunopathology. The liposomes are targeted to and selectively taken up by the affected tissue.

[0183] Dosing regimens vary depending on several factors, including the serum or tissue metabolic rate of the therapeutic antibody, the severity of the symptoms, the immunogenicity of the therapeutic antibody, and the accessibility of target cells within the biological matrix. Preferably, the dosing regimen delivers enough therapeutic antibody to produce improvement in the target condition while simultaneously minimizing undesirable side effects. Thus, the amount of biologic agent delivered will vary, in part, depending on the particular therapeutic antibody and the severity of the condition being treated. Guidance for selecting appropriate doses of therapeutic antibodies is available (e.g., Wawrzynczak (1996) Antibody Therapy, Bios Scientific Pub. Ltd, Oxfordshire, UK; Kresina (ed.) (1991) Monoclonal Antibodies, Cytokines and Arthritis, Marcel Dekker, New York, NY; Bach (ed.) (1993) Monoclonal Antibodies and Peptide Therapy in Autoimmune Diseases, Marcel Dekker, New York, NY; Baert, et al. (2003) New Engl. J. Med. 348:601-608; Milgrom et al. (1999) New Engl. J. Med. 341:1966-1973; Slamon et al. (2001) New Engl. J. Med. 344:783-792; Beniaminovitz et al. (2000) New Engl. J. Med. 342:613-619; Ghosh et al. (2003) New Engl. J. Med. 348:24-32; Lipsky et al. (2000) New Engl. J. Med. 343:1594-1602).

[0184] The appropriate dose can be determined by the clinician, for example, using parameters or factors known or suspected in the art to affect treatment. Generally, the dose is initiated somewhat below the optimal dose and then increased by small increments until a desired or optimal effect is achieved, relative to any negative side effects. Important diagnostic indicators include, for example, indicators of inflammatory symptoms or the amount of inflammatory cytokines produced. Generally, it is desirable that the biological agent used be derived from the same species as the animal targeted for treatment, thereby minimizing any immune response to the reagent. For human subjects, for example, chimeric, humanized, and fully human antibodies may be desirable.

[0185] The antibody or antigen-binding fragment thereof can be provided by continuous infusion or by doses administered, for example, 1 to 7 times per week, weekly, biweekly, monthly, bimonthly, quarterly, semi-annually, annually, etc. Doses can be provided, for example, intravenously, subcutaneously, topically, orally, nasally, rectally, intramuscularly, intracerebrally, intraspinally, or by inhalation. In some embodiments, the antibody is administered every 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks. In some embodiments, the antibody is administered every 4 weeks. In some embodiments, the antibody is administered every 5 weeks. In some embodiments, the antibody is administered every 7 weeks. In some embodiments, the antibody is administered every 6 weeks. In some embodiments, the antibody is administered every 8 weeks. In some embodiments, the antibody is administered for at least 21 to 52 weeks, or more. In some embodiments, the antibody is administered on such a schedule for at least 21 weeks. In some embodiments, the antibody is administered on such a schedule for at least 24 weeks. In some embodiments, the antibody is administered on such a schedule for at least 32 weeks. In some embodiments, the antibody is administered on such a schedule for at least 36 weeks. In some embodiments, the antibody is administered on such a schedule for at least 40 weeks. In some embodiments, the antibody is administered on such a schedule for at least 42 weeks. In some embodiments, the antibody is administered once (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered twice (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered three times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered four times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered five times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered six times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered seven times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered eight times (e.g., by infusion or continuous injection).In some embodiments, the antibody is administered 9 times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered 10 times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered 11 times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered 12 times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered 13 times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered 14 times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered 15 times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered 16 times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered 17 times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered 18 times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered 19 times (e.g., by infusion or continuous injection). In some embodiments, the antibody is administered 20 times (e.g., by infusion or continuous injection). When the antibody is administered more than once, the antibody can be administered according to a schedule such as the schedules provided herein.

[0186] The total weekly dose can be as provided herein. In some embodiments, the total weekly dose is at least 0.05 μg / kg body weight, more typically at least 0.2 μg / kg, 0.5 μg / kg, 1 μg / kg, 10 μg / kg, 100 μg / kg, 0.25 mg / kg, 1.0 mg / kg, 2.0 mg / kg, 5.0 mg / mL, 10 mg / kg, 25 mg / kg, 50 mg / kg, or more (e.g., Yang, et al. (2003) New Engl. J. Med. 349:427-434; Herold, et al. (2002) New Engl. J. Med. 346:1692-1698; Liu, et al. (1999) J. Neurol. Neurosurg. Psych. 67:451-456; Portielji, et al. (2003) Cancer Immunol.Immunother.52:133-1 44). Doses can also be provided to achieve a predetermined target concentration of antibody in the subject's serum, e.g., 0.1, 0.3, 1, 3, 10, 30, 100, 300 μg / mL.

[0187] In some embodiments, the antibody has a serum concentration in the subject of at least or about 10 μg / mL, or 20 μg / mL, 50 μg / mL, 70 μg / mL, 75 μg / mL, 80 μg / mL, 85 μg / mL, 90 μg / mL, 95 μg / mL, 100 μg / mL, or 105 μg / mL at least 1, 2, or 3 weeks after administration.

[0188] In some embodiments, a dose of 20 mg / kg IV is administered. In some embodiments, a dose is used to achieve a Cmin of 133 μg / mL after about 5 weeks. In some embodiments, a dose of antibody is administered to achieve a Cmin of 102 μg / mL after 6 weeks. In some embodiments, the antibody dose is as provided herein, such as a 10 mg / mg loading dose with subsequent doses equal to or less than the loading dose. In some embodiments, the antibody is administered at a dose as provided herein to achieve a Cmin of at least or about 100 μg / mL.

[0189] As used herein, "inhibiting" or "treating" or "treatment" includes slowing the progression of symptoms associated with a disorder and / or reducing the severity of symptoms of such a disorder. These terms further include alleviating existing, uncontrolled, or unwanted symptoms, preventing further symptoms, and preventing the underlying causes of such symptoms. Thus, these terms refer to the production of a beneficial outcome in a vertebrate subject having a disorder, disease, or condition, or at risk of developing such a disorder, disease, or condition.

[0190] As used herein, the terms "therapeutically effective amount," "therapeutically effective dose," and "effective amount" refer to an amount of an antibody or antigen-binding fragment thereof that, when administered alone or together with an additional therapeutic agent to a cell, tissue, or subject, is effective to cause a measurable improvement in one or more symptoms of a disease or condition, or in the progression of such a disease or condition. A therapeutically effective dose further refers to an amount of the binding compound sufficient to result in at least partial relief of a symptom, e.g., treatment, cure, prevention, or alleviation of an associated medical condition, or an increase in the rate of treatment, cure, prevention, or alleviation of such a condition. When applied to an individual active ingredient administered alone, a therapeutically effective dose refers to that ingredient alone. When applied to a combination, a therapeutically effective dose refers to the combined amount of the active ingredients that produces a therapeutic effect, whether the combination is sequential or simultaneous. An effective amount of a therapeutic agent results in an improvement in a diagnostic indicator or parameter of at least 10%; usually at least 20%; preferably at least about 30%; more preferably at least 40%, and most preferably at least 50%. An effective amount may also result in an improvement in subjective indicators, where subjective indicators are used to assess disease severity. In some embodiments, an amount is a therapeutically effective amount if it is an amount that can be used to treat or alleviate a condition provided herein.

[0191] The term "subject," as used throughout, includes any living organism, such as a mammal (e.g., rat, mouse, dog, cat, rabbit), and an animal, including, for example, a human. A subject can also be referred to as a patient. In some embodiments, a subject is a subject in need. A subject "in need" means a subject who has been identified as needing treatment for a condition in need of treatment and who is being treated for the specific purpose of treating such condition. The condition can be, for example, any of the conditions described herein.

[0192] On the other hand, isolated antibodies that bind to an epitope of the IGF-1R protein or other proteins described herein and exhibit IGF-1R inhibitory or therapeutic activity in vitro and / or in vivo, or antigen-binding fragments thereof capable of inhibiting IGF-1R function, are suitable as therapeutic agents for treating IGF-1R-related conditions in both humans and animals. These conditions include thyroid eye disease. Accordingly, methods for treating such conditions are also provided, comprising administering an antibody or antigen-binding fragment thereof to a subject with such a condition.

[0193] In some embodiments, the methods involve administering a therapeutically or prophylactically effective amount of one or more monoclonal antibodies or antigen-binding fragments of the antibodies described herein to a subject who is susceptible or who exhibits a condition for which IGF-1R is known or suspected to be the cause of the observed condition. Any active form of the antibody can be administered, including, but not limited to, scFv, Fab, and F(ab')2 fragments, as well as other forms of the antibody provided herein.

[0194] As used herein, an IGF-1R-associated condition refers to a condition caused by the regulation of IGF-1R, including, but not limited to, thyroid eye disease and other conditions provided herein.

[0195] In some embodiments, the antibodies used are compatible with the recipient species so that an immune response to the MAb does not result in an unacceptably short circulating half-life or induce an immune response to the MAb in the subject.

[0196] Treatment of an individual may include administering a therapeutically effective amount of an antibody described herein. The antibody can be provided in a kit such as those provided herein. The antibody can be used or administered alone or in combination with another therapeutic, analgesic, or diagnostic agent, such as those provided herein. When providing a patient with an antibody or fragment thereof capable of binding to IGF-1R, or an antibody capable of protecting against IGF-1R, a pathology in the recipient patient, the dose of the administered agent will vary depending on factors such as the patient's age, weight, height, sex, general condition, and previous medical history.

[0197] Antibodies capable of treating conditions associated with IGF-1R activity or usable to treat IGF-1R-related conditions can be provided to a subject in an amount sufficient to affect a reduction, amelioration, or alleviation of an IGF-1R-related symptom or condition, such as thyroid eye disease.

[0198] Thus, in some embodiments, a method for treating a subject suffering from an IGF-1R-mediated disorder is provided. In some embodiments, the method comprises administering a pharmaceutical composition comprising an antibody or antigen-binding fragment thereof provided herein. In some embodiments, the disorder is thyroid eye disease. As provided herein, the antibody or antigen-binding fragment thereof can be administered together with other therapeutic agents. These can be administered simultaneously or sequentially.

[0199] In some embodiments, the antibody or antigen-binding fragment thereof can be used to treat thyroid eye disease. In some embodiments, the antibody or antigen-binding fragment thereof can be used to treat or reduce the severity of or symptoms of thyroid-associated eye disease (TAO).

[0200] In some embodiments, there is provided a method or use for reducing exophthalmos in the eye in a subject suffering from thyroid-associated eye disease (TAO).

[0201] In some embodiments, the subject has been previously treated with an antibody different from those provided herein.

[0202] In some embodiments, the method or use provides for a Clinical Activity Score (CAS) in a subject having or suspected of having Thyroid Associated Eye Disease (TAO).

[0203] In some embodiments, a method or use is provided to a) reduce exophthalmos by at least 2 mm, and b) lower the Clinical Activity Score (CAS).

[0204] As used herein, the term Clinical Activity Score (CAS) refers to the scoring procedure described in Table 2. According to this procedure, the presence of each of the parameters described in the table below will award one point. The sum of all points defines the clinical activity and provides the CAS, where 0 or 1 constitutes inactive disease and 7 constitutes severe active eye disease. [Table 6]

[0205] As shown in Table 2, the CAS consists of seven components: spontaneous retrobulbar pain, pain when attempting to move the eye (staring up, side-to-side, and down; sometimes referred to as "gazing-induced orbital pain"), conjunctival redness, eyelid redness, caruncle / fold swelling, and eyelid swelling. Each component is scored as present (1 point) or absent (0 point). The score for each efficacy assessment is the sum of all present items, giving a range of 0 to 7, with 0 or 1 constituting inactive disease and 7 constituting severe active eye disease. A change of more than 2 points is considered clinically significant.

[0206] Item 1, spontaneous orbital pain, can be pain or pressure in or behind the eyeball. This pain can be caused by increased intraorbital pressure when the orbital tissue volume increases due to excessive synthesis of extracellular matrix, fluid accumulation, and cellular infiltration and swelling. Item 2, gaze-induced orbital pain, can be pain within the eye when looking or attempting to look up, down, or to the side, i.e., pain associated with upward, downward, or horizontal eye movement or when attempting to move the eye. This type of pain is caused by the stretching of inflamed muscle(s), especially when attempting to gaze upward. It is possible that "stretching pain" is not elicited by pressing the eyeball with a finger, as it is predicted to be a sign of increased intraorbital pressure. Neither type of pain may be relieved after anti-inflammatory treatment. Therefore, these pains are directly related to autoimmune inflammation in the orbit and are therefore useful for assessing TAO activity.

[0207] Swelling in TAO is seen as chemosis (edema of the conjunctiva), item 6 in Table 1, and swelling of the caruncle and / or semilunar folds. Both are signs of TAO activity. Swollen eyelids can be caused by edema, fat herniation through the orbital septum, or fibrous degeneration. In addition to swelling, other symptoms indicative of active TAO include redness and / or pain in the conjunctiva, eyelid, caruncle, and / or semilunar folds.

[0208] In some embodiments, the subject being treated has a reduction in exophthalmos of at least 2 mm, in some embodiments, the subject being treated has a reduction in exophthalmos of at least 3 mm, in some embodiments, the subject being treated has a reduction in exophthalmos of at least 4 mm.

[0209] In some embodiments, a treated subject's Clinical Activity Score (CAS) is reduced by at least 2 points. In some embodiments, the subject's Clinical Activity Score (CAS) is reduced to 1. In some embodiments, the subject's Clinical Activity Score (CAS) is reduced to zero (0).

[0210] In some embodiments, methods are provided for treating thyroid-associated eye disease (TAO) or reducing the severity of TAO in a subject, wherein treatment with the antibody (i) reduces exophthalmos in the eye by at least 2 mm; (ii) is not accompanied by a worsening of the other (i.e., fellow) eye by 2 mm or more; and (iii) reduces CAS in the subject to (1) or zero (0).

[0211] In some embodiments, methods are provided for improving quality of life in a subject with thyroid-associated eye disease (TAO, also known as Graves' eye disease / Graves' ophthalmopathy). In some embodiments, quality of life is measured by the Graves' Eye Disease Quality of Life (GO-QoL) survey or either its visual functioning or appearance subscale. In some embodiments, treatment results in an 8-point or greater improvement in the GO-QoL. In some embodiments, treatment results in an improvement in the functioning subscale of the GO-QoL. In some embodiments, treatment results in an improvement in the appearance subscale of the GO-QoL.

[0212] In some embodiments, methods are provided for treating diplopia or reducing the severity of thyroid-associated eye disease (TAO) in a subject. In some embodiments, the diplopia is stationary diplopia. In some embodiments, the diplopia is non-stationary diplopia. In some embodiments, the diplopia is intermittent diplopia. In some embodiments, the improvement or reduction in the severity of diplopia persists for at least 20 weeks after discontinuing antibody administration. In some embodiments, the improvement or reduction in the severity of diplopia persists for at least 50 weeks after discontinuing antibody administration.

[0213] Disease severity can be measured in the following non-limiting embodiments. For example, with regard to eyelid opening, the distance between the eyelid margins is measured (in mm) with the patient in primary eye position, in a relaxed sitting position, and looking with distant fixation. With regard to eyelid swelling, the index / rating is either "absent / indeterminate," "moderate," or "severe." Eyelid redness is either absent or present. Conjunctival redness is either absent or present. In some embodiments, chemosis is either absent or present. In some embodiments, inflammation of the caruncle or folds is either absent or present. Proptosis is measured in millimeters for each patient using the same Hertel exophthalmometer and the same canthal distance. Subjective diplopia is scored from 0 to 3 (0 = no diplopia; 1 = intermittent, i.e., diplopia in primary position of gaze when tired or first waking up; 2 = variable, i.e., diplopia in extreme gaze; 3 = constant, i.e., continuous diplopia in primary or reading position). For ocular muscle involvement, the twitching movement is measured in degrees. Corneal infiltration is either absent / patchy or keratopathy / ulcer. For optic nerve involvement, i.e., best visual acuity, color vision, optic disc, and relative afferent pupillary defect, pathology is either absent or present. Additionally, if optic nerve compression is suspected, visual fields are checked. In some embodiments, patients can be classified according to the following severity categories: For example, vision-threatening thyroid eye disease: hypothyroidism-induced optic neuropathy (DON) and / or corneal breaks. This category warrants immediate intervention. Moderate-to-severe thyroid eye disease: Patients without vision-threatening disease whose eye disease sufficiently impacts daily life to justify the risk of immunosuppression (if active) or surgical intervention (if inactive). Patients with moderate-to-severe thyroid eye disease typically have one of the following: eyelid retraction of 2 mm or more, moderate or severe soft tissue involvement, exophthalmos of 3 mm or more above average for their race and gender, and variable or constant diplopia. Mild thyroid eye disease: Patients whose features of thyroid eye disease have only a minimal impact on daily life, insufficiently enough to justify immunosuppression or surgical treatment. These patients usually have only one or more of the following: slight eyelid retraction (less than 2 mm), mild soft tissue involvement, more than normal for race and sex, less than 3 mm of exophthalmos, transient diplopia or absence of diplopia, and corneal exposure that responds to lubricants.

[0214] In some embodiments, patients can be characterized by a Graves' Eye Disease Quality of Life (GO-QoL) score. In addition to exophthalmos (or proptosis) and CAS, quality of life is also assessed using the GO Quality of Life (GO-QoL) questionnaire. This questionnaire is designed to measure improved quality of life after treatment with the methods disclosed herein. In some embodiments, the questionnaire can measure the reduction or absence of side effects after treatment with an antibody, or antigen-binding fragment thereof, according to the methods disclosed herein compared to treatment with glucocorticoids. The GO-QoL is a 16-item self-administered questionnaire divided into two subsets and used to assess the perceived effect of TED by the subject on (i) their daily physical activity (as this activity relates to visual function) and (ii) psychosocial function. Quality of life is assessed using the GO QoL questionnaire. The GO-QoL questionnaire [CBTerwee et al., 1998] is completed on day 1 and at weeks 6, 12, and 24 (or PW) during the treatment period, and at months 7 and 12 (or PW) during the follow-up period. The GO-QoL is a 16-item self-administered questionnaire divided into two self-assessment subscales: one covering the impact of visual function on daily activities and the other investigating the impact on self-perceived appearance. The visual function subscale covers activities such as driving a car, walking outdoors, reading, and watching television. The appearance subscale asks subjects whether their eye disease has changed their appearance; whether it has caused others to react negatively toward them; whether it has caused social isolation; and whether it has forced them to conceal their appearance. Each subscale has eight questions that can be answered with "yes (very much)," "yes (slightly)," or "no (not at all)." Each is scored from 0 to 2, and the raw total score is then mechanically converted to a 0 to 100 scale, with 0 representing the most negative impact on quality of life and 100 representing no impact. A change of 8 points or more on the 0 to 100 scale has been shown to be clinically significant.The combined score utilizes the raw scores from both subscales and again converts these into a single 0–100 scale. The questionnaire has two self-assessment subscales. Each subscale has eight questions answered with (i) "Yes (Strongly Agree)," (ii) "Yes (Slightly)," or (iii) "No (Not at all)." Each is scored from 0 to 2, and the raw total score is then mechanically converted to a 0 to 100 scale, with 0 representing the most negative impact on quality of life and 100 representing no impact. A change of more than 8 points on the 0 to 100 scale is considered clinically significant. The combined score utilizes the raw scores from both subscales and again converts them to a single 0 to 100 scale.

[0215] Patients can also be assessed for the presence or absence of a Gorman rating of diplopia. The Gorman rating of subjective diplopia includes four categories: no diplopia (absent), diplopia when the patient is tired or awake (intermittent), diplopia upon intense staring (variable), and continuous diplopia in primary or reading position (constant). Patients are scored according to the grade of diplopia they are experiencing. An improvement of 1 grade or greater is considered clinically significant.

[0216] In some embodiments, the method comprises administering an antibody such as those provided herein. In some embodiments, the antibody is administered at a dosage of about 1 mg / kg to about 5 mg / kg of antibody as a first dose. In some embodiments, the antibody is administered at a dosage of about 5 mg / kg to about 10 mg / kg of antibody as a first dose. In some embodiments, the antibody is administered at a dosage of about 5 mg / kg to about 20 mg / kg of antibody as subsequent doses. In some embodiments, the antibody is administered in the following amounts: about 10 mg / kg of antibody as a first dose; and about 20 mg / kg of antibody in subsequent doses. In some embodiments, subsequent doses are administered every three weeks for at least 21 weeks.

[0217] In some embodiments, the antibody is administered in a pharmaceutical composition such as those provided herein. In some embodiments, the pharmaceutical composition further comprises one or more pharmaceutically active compounds for the treatment of TAO. In some embodiments, the pharmaceutical composition further comprises a corticosteroid; rituximab or other anti-CD20 antibody; tocilizumab or other anti-IL-6 antibody; or selenium, infliximab, or other anti-TNFα antibody, or a thyroid-stimulating hormone receptor (TSHR) inhibitor.

[0218] In some embodiments, the methods provided herein include administering to a subject an antibody, or antigen-binding fragment thereof, that specifically binds to and inhibits IGF-IR. In some embodiments, the antibody is as provided herein.

[0219] Kits useful for practicing the embodiments described herein are also provided. The kits include a first container containing or packaged with the antibody described above. The kits may also include other containers containing or packaged with related solutions necessary or convenient for practicing the embodiments. The containers can be made of glass, plastic, or foil and can be vials, bottles, pouches, tubes, bags, etc. The kits can also contain written information, such as procedural or analytical information for practicing the embodiments, such as the amounts of reagents contained in the first container means. The containers, along with the written information, can be in a separate container device, such as a box or bag.

[0220] In yet another aspect, the present specification provides a kit for detecting IGF-1R protein in a biological sample, the kit comprising a container holding one or more antibodies that bind to an epitope of the IGF-1R protein, and instructions for using the antibodies to bind to the IGF-1R protein to form an immunological complex, and detecting the formation of the immunological complex, such that the presence or absence of the immunological complex correlates with the presence or absence of the IGF-1R protein in the sample. An example of a vessel is a multi-well plate, which allows for simultaneous detection of IGF-1R protein in multiple samples.

[0221] In some embodiments, an antibody that binds to IGF-1R protein is provided. In some embodiments, the antibody is isolated. In some embodiments, the antibody binds specifically. In some embodiments, the antibody binds to properly folded IGF-1R protein. In some embodiments, the antibody is specific for a particular IGF-1R conformational state (open or closed). In some embodiments, the antibody binds to IGF-1R protein in the cell membrane. In some embodiments, the antibody binds to IGF-1R protein in the cell membrane within intact cells. In some embodiments, the antibody inhibits or neutralizes the function of IGF-1R protein. As used herein, the term "neutralize" means that the activity or function of the protein is inhibited. Inhibition can be complete or partial. In some embodiments, the activity or function of the protein is inhibited by at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or 99%. The percentage of inhibition can be relative to the function or activity of the protein in the absence of the antibody. In some embodiments, the antibody inhibits IGF-1R-stimulated glucose transport. In some embodiments, the antibody inhibits internalization of the IGF-1R protein.

[0222] In some embodiments, the antibody comprises a sequence provided herein, or an antigen-binding fragment thereof. In some embodiments, the antibody comprises a heavy chain CDR described herein, or an antigen-binding fragment thereof. The heavy chain can be one or more of the heavy chains described herein. In some embodiments, the antibody comprises a light chain described herein, or an antigen-binding fragment thereof.

[0223] In some embodiments, methods are provided for treating, inhibiting, or alleviating a condition associated with IGF-1R. In some embodiments, the method comprises administering to a subject an antibody described herein or a pharmaceutical composition described herein to treat, inhibit, or alleviate the condition associated with IGF-1R. In some embodiments, the condition is as described herein.

[0224] In some embodiments, a method for detecting the presence or absence of IGF-1R in a sample is provided, the method comprising contacting the sample with one or more antibodies described herein and detecting binding of the antibody to an IGF-1R antigen. In some embodiments, detected binding indicates the presence of the IGF-1R antigen, or lack of detected binding indicates the absence of the IGF-1R antigen. Detection can be performed using any known method, such as a biosensor, ELISA, sandwich assay, etc. However, in some embodiments, the method comprises detecting the presence of the protein under non-denaturing conditions. Using non-denaturing conditions, the protein of interest can be detected in its native or properly folded form.

[0225] In some embodiments, methods are provided for identifying a test antibody that binds to an epitope on an IGF-1R protein, the method comprising contacting the test antibody with the epitope on the IGF-1R protein and detecting whether the test antibody binds to the epitope. In some embodiments, detecting comprises detecting whether the test antibody binds to the protein and is competitively inhibited by an antibody comprising a sequence provided herein. In some embodiments, detecting comprises mutating one or more residues of the epitope or protein and measuring binding of the test antibody to the mutated epitope, where the mutation reduces binding of the test antibody compared to the native epitope. The test antibody is considered to bind to that epitope.

[0226] In some embodiments, methods are provided for monitoring the internalization of IGF-1R from the surface of a cell. In some embodiments, the method includes contacting a cell with an anti-IGF-1R antibody provided herein and detecting the presence of IGF-1R intracellularly or at the surface of the cell. Differences in cell surface expression can be measured, and internalization can be monitored and measured. This can be used to measure the effect of another molecule, such as a test agent, on regulating the internalization of IGF-1R protein. Thus, the antibodies provided herein can be used to identify test agents that regulate (increase or decrease) the internalization of IGF-1R protein. Test molecules that increase internalization, as measured by decreased binding of the anti-IGF-1R antibody to cell surface IGF-1R protein, can be identified according to the methods provided herein. Test molecules that decrease internalization, as measured by increased binding of the anti-IGF-1R antibody to cell surface IGF-1R protein, can be identified according to the methods provided herein. Surface expression can be measured by fluorescence, which can be done with a secondary antibody that recognizes the IGF-1R antibody or by labeling the anti-IGF-1R antibodies provided herein.

[0227] In some embodiments, methods are provided for inhibiting IGF-1 stimulated receptor phosphorylation in cells. In some embodiments, the methods include contacting the cells with an antibody provided herein or a pharmaceutical composition comprising the antibody. In some embodiments, the contacting includes administering the antibody or a pharmaceutical composition comprising the antibody to a subject. In some embodiments, the cells are intraocular cells. In some embodiments, the subject has or is at risk for thyroid eye disease (TED). In some embodiments, the antibody has an IC50 of about 0.2 nm, 0.15 nm, 0.10 nm, 0.09 nm, or less. In some embodiments, the IC50 is measured in an in vitro assay, such as the assays provided herein, including those shown in the Examples. In some embodiments, the IC50 is measured in cells that are A549 cells or HOCF cells.

[0228] In some embodiments, provided are methods for treating thyroid eye disease in a subject, the method comprising treating the subject with an antibody provided herein or a pharmaceutical composition comprising the antibody, wherein the antibody has a serum concentration in the subject of at least or about 70 μg / mL, 75 μg / mL, 80 μg / mL, 85 μg / mL, 90 μg / mL, 95 μg / mL, 100 μg / mL, or 105 μg / mL at least 1, 2, or 3 weeks after administration. In some embodiments, the serum concentration is measured after 1, 2, or 3 administrations of the antibody or a pharmaceutical composition comprising the antibody to the subject.

[0229] In some embodiments, methods are provided for inhibiting IGF-1-induced receptor autophosphorylation by at least 95%, 96%, 97%, 98%, or 99%, or 100% in a subject in need thereof. In some embodiments, the method comprises administering to the subject an antibody provided herein or a pharmaceutical composition comprising the antibody. In some embodiments, IGF-1-induced receptor autophosphorylation is inhibited in the eye or orbital region of the subject. In some embodiments, inhibiting IGF-1-induced receptor autophosphorylation treats thyroid eye disease or ameliorates a symptom described herein in the subject.

[0230] Enumerated Embodiments

[0231] In some embodiments, the embodiments provided herein include, but are not limited to: 1. a VL sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 79, or 86; a VH sequence set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 80, or 83; an LCDR sequence set forth in SEQ ID NO: 17, 18, 19, 23, 24, 25, 29, 30, 31, 35, 36, 37, 41, 42, 43, 47, 48, 49, 53, 54, 55, 59, 60, 61, or 81; HCDR sequences set forth in SEQ ID NOs: 20, 21, 22, 26, 27, 28, 32, 33, 34, 38, 39, 40, 44, 45, 46, 50, 51, 52, 56, 57, 58, 62, 63, or 64; and An antibody or antigen-binding fragment thereof, comprising any combination or variant thereof. 2. The antibody, or antigen-binding fragment thereof, of embodiment 1, wherein said antibody binds to IGF-1R. 3. The antibody of embodiment 1, wherein the antibody is a monoclonal antibody. 4. The antibody of embodiment 1, wherein the antibody is a humanized antibody. 5. The antibody of embodiment 1, wherein the antibody is an scFv antibody. 6. The antibody or antigen-binding fragment thereof is selected from the group consisting of V, VIII, VIII, VIV ... L 6. The antibody of any one of embodiments 1 to 5, comprising a peptide, or a variant of any of these. 7. The antibody or antigen-binding fragment thereof is selected from the group consisting of V, ... H 7. The antibody of any one of embodiments 1 to 6, comprising a peptide, or a variant of any of these. 8. An antibody or antigen-binding fragment thereof, wherein the antibody or antibody fragment has (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 20, 26, 32, 38, 44, 50, or 56; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 21, 27, 33, 39, 45, 51, or 57; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 22, 28, 34, 40, 46, 52, or 58; or a variant of any of the foregoing. the antibody or antigen-binding fragment thereof, comprising: (i) a heavy chain variable region; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 17, 23, 29, 35, 41, 47, or 53; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 18, 24, 30, 36, 42, 48, or 54; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 19, 25, 31, 37, 43, 49, 55, or 81; or a variant of any of the foregoing. 9. An antibody or antigen-binding fragment thereof, comprising: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 20; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 21; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 22; or any variant thereof; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 17; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 18; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 19; or any variant thereof. 10. An antibody or antigen-binding fragment thereof, wherein the antibody or antibody fragment comprises: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 26; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 27; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 28; or a variant of any of the foregoing; and (ii) a light chain CDR1, CDR2 and a light chain variable region comprising a CDR1 sequence of SEQ ID NO: 23; a light chain CDR2 sequence of SEQ ID NO: 24; and a light chain CDR3 sequence of SEQ ID NO: 25; or a variant of any of the foregoing. 11. An antibody or antigen-binding fragment thereof, comprising: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 32; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 33; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 34; or any variant thereof; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 29; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 30; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 31; or any variant thereof. 12. An antibody or antigen-binding fragment thereof, comprising: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 38; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 39; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 40; or any variant thereof; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 35; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 36; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 37; or any variant thereof. 13. An antibody or antigen-binding fragment thereof, comprising: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 44; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 45; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 46; or any variant thereof; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 41; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 42; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 43; or any variant thereof. 14. An antibody or antigen-binding fragment thereof, comprising: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 50; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 51; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 52; or any variant thereof; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 47; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 48; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 49; or any variant thereof. 15. An antibody or antigen-binding fragment thereof, comprising: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 56; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 57; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 58; or any variant thereof; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 53; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 54; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 55; or any variant thereof. 16. An antibody or antigen-binding fragment thereof, wherein the antibody or antibody fragment comprises: (i) a heavy chain C The antibody or antigen-binding fragment thereof, comprising: (i) a heavy chain variable region comprising CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 62; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 63; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 64; or any variant thereof; and (ii) a light chain variable region comprising CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 59; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 60; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 61; or any variant thereof. 17. An antibody or antigen-binding fragment thereof, comprising: (i) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 38; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 39; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 40; or any variant thereof; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 35; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 36; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 81; or any variant thereof. 18. The antibody of any one of embodiments 6 to 17, wherein the heavy chain variable region and the light chain variable region are not linked by a linker. 19. The antibody of any one of embodiments 6 to 17, wherein the heavy chain variable region and the light chain variable region are linked by a peptide linker. 20. The peptide linker is (GGGGS) n (SEQ ID NO: 73); (GGGGA) n (SEQ ID NO: 74), or any combination thereof, wherein each n is independently 1 to 5. 21. The antibody of any one of embodiments 1 to 20, wherein the antibody comprises the sequence of SEQ ID NO: 65 to 72, 78, 82, or 85, or a variant thereof. 22. The antibody is selected from the group consisting of V, VB, VC, VD ... L 22. The antibody of any one of embodiments 1 to 21, comprising the sequence: 23. The antibody is selected from the group consisting of V, VB, VC, VD ... H 22. The antibody of any one of embodiments 1 to 21, comprising the sequence: 24. The isolated antibody of any one of embodiments 1 to 21, wherein the antibody comprises the sequence of SEQ ID NO: 65-72, 78, 82, or 85, or a variant thereof. 25. The antibody of any one of embodiments 1 to 24, wherein the variant has 1 to 10 substitutions, deletions, or insertions. 26. The antibody of any one of embodiments 1 to 24, wherein the variant has 1 to 10 conservative substitutions. 27. The antibody of any one of embodiments 1 to 26, wherein the variant has at least 85% homology to the sequence of SEQ ID NO: 1 to 72, 78 to 83, or 85 to 86. 28. The antibody of any one of embodiments 1 to 26, wherein the variant has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% homology to the sequence of SEQ ID NO: 1 to 72, 78 to 83, or 85 to 86. 29. The antibody of any one of embodiments 1 to 26, wherein the variant has at least 85% identity to the sequence of SEQ ID NO: 1 to 72, 78 to 83, or 85 to 86. 30. The variant has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, or more specificity to the sequence of SEQ ID NO: 1-72, 78-83, or 85-86, 27. The antibody of any one of embodiments 1 to 26, having 98% or 99% identity. 31. The antibody according to any one of embodiments 1 to 26, wherein said antibody is an scFv antibody. 32. The antibody according to any one of embodiments 1 to 26, wherein said antibody is a monoclonal antibody. 33. The antibody of any one of embodiments 1 to 26, wherein the antibody is a humanized antibody. 34. The antibody of any one of the preceding embodiments, wherein the antibody comprises an Fc region. 35. The antibody of embodiment 34, wherein the Fc region is as set forth in SEQ ID NO: 75-77, or 84. 36. The antibody of any one of the preceding embodiments, wherein the Fc region comprises a mutation that increases the half-life of the antibody when bound to the Fc region. 37. The antibody of embodiment 36, wherein the Fc region comprises an S228P, L235E, M252Y, S254T, T256E, M428L, N434S, L234F, P331S mutation, or any combination thereof. 38. The antibody of embodiment 36, wherein the Fc region comprises M252Y, S254T, and T256E mutations. 39. The antibody of embodiment 36, wherein the Fc region comprises S228P and L235E mutations. 40. The antibody of embodiment 36, wherein the Fc region comprises L234F, L235E, and P331S mutations. 41. The antibody of embodiment 36, wherein the Fc region comprises M252Y, S254T, T256E, S228P, and L235E mutations. 42. The antibody of embodiment 36, wherein the Fc region comprises S228P, L235E, M428L, and N434S mutations. 43. The antibody of embodiment 36, wherein the Fc region comprises M428L and N434S mutations. 44. The antibody of embodiment 36, wherein the Fc region comprises the following mutations: L234F, L235E, P331S, M252Y, S254T, and T256E. 45. A nucleic acid molecule encoding the antibody or antigen-binding fragment thereof according to any one of the preceding embodiments. 46. ​​A vector comprising the nucleic acid molecule of embodiment 45. 47. A cell comprising a nucleic acid molecule according to embodiment 46, or a nucleic acid comprising a vector according to embodiment 46. 48. A pharmaceutical composition comprising an antibody according to any one of embodiments 1 to 44, or a nucleic acid molecule encoding said antibody. 49. The pharmaceutical composition according to embodiment 48, wherein the composition is an injectable pharmaceutical composition. 50. A method for treating thyroid-associated eye disease (TAO) or a symptom of TAO, or reducing the severity of TAO or a symptom of TAO, comprising administering to a subject an antibody described in any one of embodiments 1 to 44, or a pharmaceutical composition comprising the antibody. 51. A method for reducing exophthalmos in a subject suffering from thyroid-associated eye disease (TAO), comprising administering to the subject an antibody of any one of embodiments 1 to 44, or a pharmaceutical composition comprising said antibody. 52. A method for treating thyroid eye disease in a subject, comprising administering to the subject an antibody according to any one of embodiments 1 to 44, or a pharmaceutical composition comprising said antibody. 53. A method for reducing the Clinical Activity Score (CAS) of Thyroid-Associated Eye Disease (TAO) in a subject, comprising administering to the subject an antibody described in any one of embodiments 1 to 44, or a pharmaceutical composition comprising said antibody. 54. A method for the treatment of a subject suffering from thyroid-associated eye disease (TAO) comprising administering to the subject an antibody according to any one of embodiments 1 to 44 or a pharmaceutical composition comprising said antibody, wherein: a) exophthalmos is reduced by at least 2 mm; and b) clinical activity score (CAS) is reduced. How to do it. 55. The method of any of embodiments 50-54, wherein exophthalmos is reduced by at least 2 mm. 56. The method of any of embodiments 50-54, wherein exophthalmos is reduced by at least 3 mm. 57. The method of any of embodiments 50-54, wherein exophthalmos is reduced by at least 4 mm. 58. The method of any of embodiments 50-54, wherein the subject's Clinical Activity Score (CAS) is reduced by at least 2 points. 59. The method of any of embodiments 50-54, wherein the subject's Clinical Activity Score (CAS) is reduced to (1). 60. The method of any of embodiments 50-54, wherein the subject's Clinical Activity Score (CAS) is reduced to zero (0). 61. A method for treating thyroid-associated eye disease (TAO) or reducing the severity of TAO in a subject, comprising administering to the subject an antibody of any one of embodiments 1-44, or a pharmaceutical composition comprising said antibody, wherein treatment with said antibody (i) reduces exophthalmos in the eye by at least 2 mm; (ii) is not accompanied by a worsening of the other (i.e., fellow) eye by 2 mm or more; and (iii) reduces CAS in the subject to (1) or zero (0). 62. A method for improving quality of life in a subject suffering from thyroid-associated eye disease (TAO, also known as Graves' eye disease / Graves' ophthalmopathy), comprising administering to the subject an antibody described in any one of embodiments 1 to 44, or a pharmaceutical composition comprising said antibody. 63. The method of embodiment 62, wherein said quality of life is measured by the Graves' Eye Disease Quality of Life (GO-QoL) survey, or either its visual functioning or appearance subscale. 64. The method of embodiment 63, wherein said treatment results in an improvement of 8 or more points in GO-QoL. 65. The method of embodiment 63, wherein said treatment results in an improvement in the functioning subscale of the GO-QoL. 66. The method of embodiment 63, wherein said treatment results in an improvement in the appearance subscale of the GO-QoL. 67. A method for treating or reducing the severity of diplopia in a subject suffering from thyroid-associated eye disease (TAO), comprising administering to the subject an antibody described in any one of embodiments 1 to 44, or a pharmaceutical composition comprising said antibody. 68. The method of embodiment 67, wherein the diplopia is stationary diplopia. 69. The method of embodiment 67, wherein the diplopia is non-stationary diplopia. 70. The method of embodiment 67, wherein the diplopia is intermittent diplopia. 71. The method of embodiment 67, wherein the improvement or reduction in the severity of diplopia persists for at least 20 weeks after discontinuing antibody administration. 72. The method of embodiment 67, wherein the improvement or reduction in the severity of diplopia persists for at least 50 weeks after discontinuing antibody administration. 73. The method of any one of embodiments 50-72, wherein the antibody is administered at a dosage of about 1 mg / kg to about 5 mg / kg of antibody as a first dose. 74. The method of any one of embodiments 50-72, wherein the antibody is administered at a dosage of about 5 mg / kg to about 10 mg / kg of antibody as a first dose. 75. The method of any one of embodiments 50-72, wherein the antibody is administered at a dose of about 5 mg / kg to about 20 mg / kg of antibody in subsequent doses. 76. The method of any one of embodiments 50-72, wherein the antibody is administered in the following amounts: about 10 mg / kg of antibody as a first dose; and about 20 mg / kg of antibody as a subsequent dose. 77. The method of embodiment 76, wherein the subsequent doses are administered every 3 weeks for at least 21 weeks. 78. The method of any one of embodiments 50 to 77, wherein the antibody or antigen-binding fragment thereof is a human antibody, a monoclonal antibody, a human monoclonal antibody, a purified antibody, a diabody, a single-chain antibody, a multispecific antibody, Fab, Fab', F(ab')2, Fv, or scFv. 79. The method of any one of embodiments 50 to 78, wherein the antibody or antigen-binding fragment thereof is administered in a pharmaceutical composition further comprising a pharmaceutically acceptable diluent or excipient or carrier. 80. The method of embodiment 79, wherein the pharmaceutical composition further comprises one or more pharmaceutically active compounds for the treatment of TAO. 81. The method of embodiment 79 or 80, wherein the pharmaceutical composition further comprises a corticosteroid; rituximab or other anti-CD20 antibody; tocilizumab or other anti-IL-6 antibody; or selenium, infliximab, or other anti-TNFα antibody, or a thyroid-stimulating hormone receptor (TSHR) inhibitor. 82. The method of any of the preceding embodiments, wherein the antibody or antigen-binding fragment thereof is administered directly to the eye, the anterior chamber of the eye, the vitreous chamber of the eye, the suprachoroidal space, or the retro-orbital space. 83. The method of embodiment 82, wherein the antibody or antigen-binding fragment thereof is administered by injection. 84. The method of embodiment 83, wherein the injection is an intravitreal injection, an intraorbital injection, a retroorbital injection, a suprachoroidal injection, or an intraluminal injection. 85. A method for increasing the internalization of IGF-1R in a cell, the method comprising contacting the cell with an antibody according to any one of embodiments 1 to 44, or a pharmaceutical composition comprising the antibody. 86. The method of embodiment 85, wherein the contacting comprises administering to the subject an antibody of any one of embodiments 1 to 44, or a pharmaceutical composition comprising said antibody. 87. The method of embodiment 86, wherein the subject has or is at risk for thyroid eye disease (TED). 88. A method for inhibiting IGF-1 stimulated receptor phosphorylation in a cell, the method comprising contacting the cell with an antibody described in any one of embodiments 1 to 44, or a pharmaceutical composition comprising the antibody. 89. The method of embodiment 88, wherein the contacting comprises administering to the subject an antibody of any one of embodiments 1 to 44, or a pharmaceutical composition comprising said antibody. 90. The method of embodiment 89, wherein the subject has or is at risk for thyroid eye disease (TED). 91. The method of any one of embodiments 88-90, wherein the antibody has an IC50 of about 0.2 nm, 0.15 nm, 0.10 nm, 0.09 nm or less. 92. The method of embodiment 91, wherein the IC50 is measured in an in vitro assay, such as the assay provided herein. 93. The method of any one of embodiments 88 to 92, wherein the cells are A549 cells or HOCF cells. 94. A method for treating thyroid eye disease in a subject, the method comprising administering to the subject an antibody described in any one of embodiments 1-44 or otherwise provided herein, or a pharmaceutical composition comprising the antibody, wherein the antibody has a serum concentration in the subject of at least or about 70 μg / mL, 75 μg / mL, 80 μg / mL, 85 μg / mL, 90 μg / mL, 95 μg / mL, 100 μg / mL, or 105 μg / mL at least 1, 2, or 3 weeks after administration. 95. The method of embodiment 94, wherein the antibody or pharmaceutical composition is administered intravenously. 96. The method of embodiment 94 or 96, wherein the antibody or pharmaceutical composition is administered at a dose of about 20 mg / kg. 97. The method of any one of embodiments 94-96, wherein the antibody or pharmaceutical composition is administered at least, or approximately, once a week, once every two weeks, once every three weeks, or once every four weeks. 98. A method for inhibiting IGF-1-induced receptor autophosphorylation in a cell by at least 95%, 96%, 97%, 98%, or 99%, or 100%, said method comprising contacting said cell with an antibody described in any one of embodiments 1 to 44 or otherwise provided herein, or a pharmaceutical composition comprising said antibody. 99. The method of embodiment 98, wherein the inhibition of IGF-1-induced receptor autophosphorylation is measured relative to induced receptor autophosphorylation in the absence of the antibody or the pharmaceutical composition. 100. The method of embodiment 98 or 99, wherein said contacting comprises administering to said subject said antibody or a pharmaceutical composition comprising said antibody. 101. The method of embodiment 100, wherein the subject has or is at risk for thyroid eye disease (TED). 102. A method for inhibiting IGF-1-induced receptor autophosphorylation by at least 95%, 96%, 97%, 98%, or 99%, or 100% in a subject in need thereof, wherein the method comprises administering to the subject an antibody according to any one of embodiments 1 to 44, or otherwise provided herein, or a pharmaceutical composition comprising the antibody. 103. The method of embodiment 102, wherein the subject has or is at risk for thyroid eye disease (TED). 104. The method of any one of embodiments 102 or 103, wherein the antibody or the pharmaceutical composition is administered intravenously. 105. The method of any one of embodiments 98 to 104, wherein the antibody comprises the CDRs of VRDN-1100. 106. The method of any one of embodiments 98 to 104, wherein the antibody comprises the CDRs of VRDN-1100 or the CDRs of VRDN-2700. 107. An isolated antibody, comprising a light chain having the amino acid sequence of SEQ ID NO: 3, and a heavy chain comprising the amino acid sequence of SEQ ID NO: 83. 108. An isolated antibody comprising a light chain variable region having the amino acid sequence of SEQ ID NO: 13 and a heavy chain variable region having the amino acid sequence of SEQ ID NO: 14. 109. The isolated antibody of embodiment 108, wherein the antibody comprises a light chain having the amino acid sequence of SEQ ID NO: 93 and a heavy chain having the amino acid sequence of SEQ ID NO: 92. 110. The isolated antibody of embodiment 108, wherein the antibody comprises a light chain having the amino acid sequence of SEQ ID NO: 93 and a heavy chain having the amino acid sequence of SEQ ID NO: 94. 111. The isolated antibody of embodiment 108, wherein the antibody comprises a light chain having the amino acid sequence of SEQ ID NO: 93 and a heavy chain having the amino acid sequence of SEQ ID NO: 95. 112. A pharmaceutical composition comprising an antibody according to any one of embodiments 107 to 111. 113. A pharmaceutical composition suitable for intravenous administration, comprising an antibody according to any one of embodiments 107 to 111. 114. A pharmaceutical composition suitable for subcutaneous administration, comprising an antibody according to any one of embodiments 107 to 111. 115. A method for treating thyroid eye disease in a subject, said method comprising administering a pharmaceutical composition comprising the antibody of any one of embodiments 107 to 111. 116. The method of embodiment 115, wherein the pharmaceutical composition is administered intravenously. 117. The method of embodiment 115, wherein the pharmaceutical composition is administered subcutaneously. 118. A method for treating thyroid-associated eye disease (TAO) or TAO, comprising administering to a subject an antibody according to any one of embodiments 107 to 111, or a pharmaceutical composition comprising said antibody. or a method for reducing the severity of TAO or the symptoms of TAO. 119. A method for reducing exophthalmos in a subject suffering from thyroid-associated eye disease (TAO), comprising administering to the subject an antibody of any one of embodiments 107 to 111, or a pharmaceutical composition comprising said antibody. 120. A method for treating thyroid eye disease in a subject, comprising administering to the subject an antibody according to any one of embodiments 107 to 111, or a pharmaceutical composition comprising said antibody. 121. A method for reducing the Clinical Activity Score (CAS) of Thyroid-Associated Eye Disease (TAO) in a subject, comprising administering to the subject an antibody described in any one of embodiments 107 to 111, or a pharmaceutical composition comprising said antibody. 122. A method for a) reducing exophthalmos by at least 2 mm and b) lowering Clinical Activity Score (CAS) in a subject suffering from thyroid-associated eye disease (TAO), comprising administering to the subject an antibody described in any one of embodiments 107 to 111, or a pharmaceutical composition comprising said antibody. 123. The method of any of embodiments 118-122, wherein exophthalmos is reduced by at least 2 mm. 124. The method of any of embodiments 118-122, wherein exophthalmos is reduced by at least 3 mm. 125. The method of any of embodiments 118-122, wherein exophthalmos is reduced by at least 4 mm. 126. The method of any of embodiments 118-122, wherein the subject's Clinical Activity Score (CAS) is reduced by at least 2 points. 127. The method of any of embodiments 118-122, wherein the subject's Clinical Activity Score (CAS) is reduced to (1). 128. The method of any of embodiments 118-122, wherein the subject's Clinical Activity Score (CAS) is reduced to zero (0). 129. A method for treating thyroid-associated eye disease (TAO) or reducing the severity of TAO in a subject, comprising administering to the subject an antibody of any one of embodiments 107 to 111, or a pharmaceutical composition comprising the antibody, wherein treatment with the antibody (i) reduces exophthalmos in the eye by at least 2 mm; (ii) is not accompanied by a worsening of the other (i.e., fellow) eye by 2 mm or more; and (iii) reduces CAS in the subject to (1) or zero (0). 130. A method for improving quality of life in a subject suffering from thyroid-associated eye disease (TAO, also known as Graves' eye disease / Graves' ophthalmopathy), comprising administering to the subject an antibody described in any one of embodiments 107 to 111, or a pharmaceutical composition comprising said antibody. 131. The method of embodiment 130, wherein said quality of life is measured by the Graves' Eye Disease Quality of Life (GO-QoL) survey, or either its visual functioning or appearance subscale. 132. The method of embodiment 130, wherein said treatment results in an improvement of 8 or more points in GO-QoL. 133. The method described in embodiment 130, wherein said treatment results in an improvement in the functioning subscale of the GO-QoL. 134. The method described in embodiment 130, wherein said treatment results in an improvement in the appearance subscale of the GO-QoL. 135. A method for treating or reducing the severity of diplopia in a subject suffering from thyroid-associated eye disease (TAO), comprising administering to the subject an antibody according to any one of embodiments 107 to 111, or a pharmaceutical composition comprising said antibody. 136. The method of embodiment 135, wherein the diplopia is stationary diplopia. 137. The method of embodiment 135, wherein the diplopia is non-stationary diplopia. 138. The method of embodiment 135, wherein the diplopia is intermittent diplopia. 139. Improvement or reduction in the severity of diplopia persists for at least 20 weeks after discontinuing antibody administration. 136. The method of embodiment 135, wherein the method is persistent. 140. The method of embodiment 135, wherein the improvement or reduction in the severity of diplopia persists for at least 50 weeks after discontinuing antibody administration. 141. The method of any one of embodiments 115-140, wherein the antibody is administered at a dosage of about 1 mg / kg to about 5 mg / kg of antibody as a first dose. 142. The method of any one of embodiments 115-140, wherein the antibody is administered at a dosage of about 5 mg / kg to about 10 mg / kg of antibody as a first dose. 143. The method of any one of embodiments 115-140, wherein the antibody is administered at a dose of about 5 mg / kg to about 20 mg / kg of antibody in subsequent doses. 144. The method of any one of embodiments 115-140, wherein the antibody is administered in the following amounts: about 10 mg / kg of antibody as a first dose; and about 20 mg / kg of antibody as a subsequent dose. 145. The method of embodiment 144, wherein the subsequent doses are administered every 3 weeks for at least 21 weeks. 146. The method of any one of embodiments 115-140, wherein the antibody is administered in a pharmaceutical composition comprising a pharmaceutically acceptable diluent, excipient, or carrier. 147. The method of embodiment 146, wherein the pharmaceutical composition further comprises one or more pharmaceutically active compounds for the treatment of TAO. 148. The method of embodiment 146 or 147, wherein the pharmaceutical composition further comprises a corticosteroid; rituximab or other anti-CD20 antibody; tocilizumab or other anti-IL-6 antibody; or selenium, infliximab, or other anti-TNFα antibody, or a thyroid-stimulating hormone receptor (TSHR) inhibitor. 149. A method for increasing the internalization of IGF-1R in a cell, the method comprising contacting the cell with an antibody according to any one of embodiments 107 to 111, or a pharmaceutical composition comprising the antibody. 150. The method of embodiment 149, wherein said contacting comprises administering to the subject said antibody or a pharmaceutical composition comprising said antibody. 151. The method of embodiment 150, wherein the subject has or is at risk for thyroid eye disease (TED). 152. A method for inhibiting IGF-1 stimulated receptor phosphorylation in a cell, the method comprising contacting the cell with an antibody according to any one of embodiments 107 to 111, or a pharmaceutical composition comprising the antibody. 153. The method of embodiment 152, wherein the contacting comprises administering to the subject an antibody of any one of embodiments 1 to 44, or a pharmaceutical composition comprising said antibody. 154. The method of embodiment 153, wherein the subject has or is at risk for thyroid eye disease (TED). 155. The method of embodiment 153 or 154, wherein the antibody has an IC50 of about 0.2 nm, 0.15 nm, 0.10 nm, 0.09 nm or less. 156. The method of embodiment 155, wherein the IC50 is measured in an in vitro assay, such as the assay provided herein. 157. The method of any one of embodiments 152 to 157, wherein the cells are A549 cells or HOCF cells. 158. A method for treating thyroid eye disease in a subject, the method comprising administering to the subject an antibody of any one of embodiments 107 to 111, or a pharmaceutical composition comprising the antibody, wherein the antibody has a serum concentration in the subject of at least or about 70 μg / mL, 75 μg / mL, 80 μg / mL, 85 μg / mL, 90 μg / mL, 95 μg / mL, 100 μg / mL, or 105 μg / mL at least 1, 2, or 3 weeks after administration. 159. The method of embodiment 158, wherein the antibody or pharmaceutical composition is administered intravenously. Law. 160. The method of embodiment 158 ​​or 159, wherein the antibody or pharmaceutical composition is administered at about 1 mg / kg to about 5 mg / kg (mg antibody / kg subject), or about 5 mg / kg to about 10 mg / kg of antibody, or about 5 mg / kg to about 20 mg / kg, in a first dose or subsequent doses. 161. The method of any one of embodiments 158-160, wherein the antibody is administered in the following amounts: about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mg / kg of antibody as a first dose; and about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg / kg of antibody in subsequent doses. 162. The method of any one of embodiments 158-161, wherein the antibody or pharmaceutical composition is administered at least, or approximately, once a week, once every two weeks, once every three weeks, or once every four weeks. 163. A method for inhibiting IGF-1-induced receptor autophosphorylation by at least 95%, 96%, 97%, 98%, or 99%, or 100% in a subject in need thereof, said method comprising administering to said subject an antibody according to any one of embodiments 107 to 111, or a pharmaceutical composition comprising said antibody. 164. A pharmaceutical composition comprising an antibody for treating thyroid eye disease in a subject, wherein the antibody comprises a light chain variable region having the amino acid sequence of SEQ ID NO: 13 and a heavy chain variable region having the amino acid sequence of SEQ ID NO: 14. 165. The pharmaceutical composition of embodiment 164, wherein the antibody comprises an Fc region having M428L and N434S substitutions. 166. The pharmaceutical composition of embodiment 164, wherein the antibody comprises an Fc region with M428L, N434S, M252Y, S254T, and T256E substitutions. 167. The pharmaceutical composition of embodiment 164, wherein the antibody comprises an Fc region with M252Y, S254T, and T256E substitutions. 168. The pharmaceutical composition of embodiment 164, wherein the antibody comprises a light chain having the amino acid sequence of SEQ ID NO: 93 and a heavy chain having the amino acid sequence of SEQ ID NO: 92. 169. The pharmaceutical composition of embodiment 164, wherein the antibody comprises a light chain having the amino acid sequence of SEQ ID NO: 93 and a heavy chain having the amino acid sequence of SEQ ID NO: 94. 170. The pharmaceutical composition of embodiment 164, wherein the antibody comprises a light chain having the amino acid sequence of SEQ ID NO: 93 and a heavy chain having the amino acid sequence of SEQ ID NO: 95. 171. A method for treating thyroid eye disease in a subject, said method comprising administering a pharmaceutical composition comprising the antibody of any one of embodiments 164 to 170. 172. The method of embodiment 171, wherein the pharmaceutical composition is administered intravenously. 173. The method of embodiment 171, wherein the pharmaceutical composition is administered subcutaneously. 174. A method for treating thyroid-associated eye disease (TAO), or a symptom of TAO, or reducing the severity of TAO, or a symptom of TAO, said method comprising administering to a subject a pharmaceutical composition described in any one of embodiments 164 to 170. 175. A method for reducing exophthalmos in a subject suffering from thyroid-associated eye disease (TAO), said method comprising administering to the subject a pharmaceutical composition according to any one of embodiments 164 to 170. 176. A method for treating thyroid eye disease in a subject, said method comprising administering to the subject a pharmaceutical composition of any one of embodiments 2-4. 177. A method for reducing the Clinical Activity Score (CAS) of Thyroid-Associated Eye Disease (TAO) in a subject, the method comprising administering to the subject a pharmaceutical composition described in any one of embodiments 164 to 170. 178. A method for a) reducing exophthalmos by at least 2 mm and b) reducing clinical activity score (CAS) in a subject with thyroid-associated eye disease (TAO), comprising: The method comprises administering to a subject a pharmaceutical composition described in any one of embodiments 164 to 170. 179. The method of any of embodiments 174-178, wherein exophthalmos is reduced by at least 2 mm. 180. The method of any of embodiments 174-178, wherein exophthalmos is reduced by at least 3 mm. 181. The method according to any of embodiments 174-178, wherein exophthalmos is reduced by at least 4 mm. 182. The method of any of embodiments 174-178, wherein the subject's Clinical Activity Score (CAS) is reduced by at least 2 points. 183. The method of any of embodiments 174-178, wherein the subject's Clinical Activity Score (CAS) is reduced to (1). 184. The method of any of embodiments 174-178, wherein the subject's Clinical Activity Score (CAS) is reduced to zero (0). 185. A method for treating thyroid-associated eye disease (TAO) or reducing the severity of TAO in a subject, comprising administering to the subject the pharmaceutical composition of any one of embodiments 164-170, wherein treatment with the antibody (i) reduces exophthalmos in the eye by at least 2 mm; (ii) is not accompanied by a worsening of the other (i.e., fellow) eye by 2 mm or more; and (iii) reduces CAS in the subject to (1) or zero (0). 186. A method for improving the quality of life in a subject suffering from thyroid-associated eye disease (TAO, also known as Graves' eye disease / Graves' ophthalmopathy), the method comprising administering to the subject a pharmaceutical composition described in any one of embodiments 164 to 170. 187. The method of embodiment 186, wherein said quality of life is measured by the Graves' Eye Disease Quality of Life (GO-QoL) survey, or either its visual functioning or appearance subscale. 188. The method of embodiment 186, wherein said treatment results in an improvement of 8 or more points in GO-QoL. 189. The method described in embodiment 186, wherein said treatment results in an improvement in the functioning subscale of the GO-QoL. 190. The method of embodiment 186, wherein said treatment results in an improvement in the appearance subscale of the GO-QoL. 191. A method for treating diplopia or reducing the severity of diplopia in a subject suffering from thyroid-associated eye disease (TAO), said method comprising administering to the subject a pharmaceutical composition described in any one of embodiments 164-170. 192. The method of embodiment 191, wherein the diplopia is stationary diplopia. 193. The method of embodiment 191, wherein the diplopia is non-stationary diplopia. 194. The method of embodiment 191, wherein the diplopia is intermittent diplopia. 195. The method of embodiment 191, wherein the improvement or reduction in the severity of diplopia persists for at least 20 weeks after discontinuing antibody administration. 196. The method of embodiment 191, wherein the improvement or reduction in the severity of diplopia persists for at least 50 weeks after discontinuing antibody administration. 197. The method of any one of embodiments 171-196, wherein the pharmaceutical composition is administered at a first dose of about 1 mg / kg to about 5 mg / kg, about 5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 20 mg / kg, about 20 mg / kg to about 30 mg / kg, about 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, or about 30 mg / kg of antibody. 198. The method of any one of embodiments 171-196, wherein the pharmaceutical composition is administered at a dosage of about 10 mg / kg to about 20 mg / kg of antibody as a first dose. 199. The pharmaceutical composition is administered in a subsequent dose of about 1 mg / kg to about 10 mg / kg, about 2 mg 197. The method of any one of embodiments 171-196, wherein the antibody is administered at a dose of from about 5 mg / kg to about 5 mg / kg, or from about 5 mg / kg to about 20 mg / kg. 200. The method of any one of embodiments 171-196, wherein the pharmaceutical composition is administered in the following amounts: about 10 mg / kg of antibody as a first dose; and about 20 mg / kg of antibody as a subsequent dose. 201. The method of embodiment 200, wherein the subsequent doses are administered every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, or every 8 weeks for at least 21 to 52 weeks, or more. 202. A method for increasing the internalization of IGF-1R in a cell, said method comprising contacting said cell with a pharmaceutical composition according to any one of embodiments 164 to 170. 203. The method of embodiment 202, wherein said contacting comprises administering to the subject a pharmaceutical composition of any one of embodiments 164 to 170. 204. The method of embodiment 203, wherein the subject has or is at risk for thyroid eye disease (TED). 205. A method for inhibiting IGF-1 stimulated receptor phosphorylation in a cell, said method comprising contacting said cell with a pharmaceutical composition according to any one of embodiments 164 to 170. 206. The method of embodiment 205, wherein said contacting comprises administering to the subject a pharmaceutical composition of any one of embodiments 164 to 170. 207. The method of embodiment 206, wherein the subject has or is at risk for thyroid eye disease (TED). 208. The method of any one of embodiments 205-207, wherein the antibody has an IC50 of about 0.2 nm, 0.15 nm, 0.10 nm, 0.09 nm or less. 209. A method for treating thyroid eye disease in a subject, the method comprising administering to the subject the pharmaceutical composition of any one of embodiments 164-170, wherein the antibody has a serum concentration in the subject of at least or about 10 μg / mL, or 20 μg / mL, 50 μg / mL, 70 μg / mL, 75 μg / mL, 80 μg / mL, 85 μg / mL, 90 μg / mL, 95 μg / mL, 100 μg / mL, or 105 μg / mL at least 1, 2, or 3 weeks after administration. 210. The method of embodiment 209, wherein said pharmaceutical composition is administered intravenously or subcutaneously. 211. An isolated antibody, comprising a light chain having the amino acid sequence of SEQ ID NO: 3, and a heavy chain comprising the amino acid sequence of SEQ ID NO: 83. 212. An isolated antibody, comprising a variable light chain comprising the sequence of SEQ ID NO: 98, and a variable heavy chain comprising the sequence of SEQ ID NO: 99, and an Fc region comprising M252Y, S254T, and T256E mutations. 213. An isolated antibody, comprising a variable light chain comprising the sequence of SEQ ID NO: 98, and a variable heavy chain comprising the sequence of SEQ ID NO: 99, and an Fc region comprising M428L and N434S mutations. 214. A pharmaceutical composition comprising an antibody according to any one of embodiments 211 to 213. 215. A pharmaceutical composition suitable for intravenous administration, comprising an antibody according to any one of embodiments 211 to 213. 216. A pharmaceutical composition suitable for subcutaneous administration, comprising an antibody according to any one of embodiments 211 to 213. 217. A method for treating thyroid eye disease in a subject, said method comprising administering a pharmaceutical composition comprising the antibody of any one of embodiments 211 to 213. 218. The method of embodiment 217, wherein the pharmaceutical composition is administered intravenously. 219. The method of embodiment 217, wherein the pharmaceutical composition is administered subcutaneously. 220. A method for treating thyroid-associated eye disease (TAO) or a symptom of TAO, or reducing the severity of TAO or a symptom of TAO, comprising administering to a subject an antibody of any one of embodiments 211 to 213, or a pharmaceutical composition comprising said antibody. 221. A method for reducing exophthalmos in a subject suffering from thyroid-associated eye disease (TAO), comprising administering to the subject an antibody of any one of embodiments 1 to 44, or a pharmaceutical composition comprising said antibody. 222. A method for treating thyroid eye disease in a subject, comprising administering to the subject an antibody according to any one of embodiments 211 to 213, or a pharmaceutical composition comprising said antibody. 223. A method for reducing the Clinical Activity Score (CAS) of Thyroid-Associated Eye Disease (TAO) in a subject, comprising administering to the subject an antibody according to any one of embodiments 211 to 213, or a pharmaceutical composition comprising said antibody. 224. A method for a) reducing exophthalmos by at least 2 mm and b) lowering clinical activity score (CAS) in a subject suffering from thyroid-associated eye disease (TAO), comprising administering to the subject an antibody described in any one of embodiments 211 to 213, or a pharmaceutical composition comprising said antibody. 225. The method of any of embodiments 220-224, wherein exophthalmos is reduced by at least 2 mm. 226. The method of any of embodiments 220-224, wherein exophthalmos is reduced by at least 3 mm. 227. The method of any of embodiments 220-224, wherein exophthalmos is reduced by at least 4 mm. 228. The method of any of embodiments 220-224, wherein the subject's Clinical Activity Score (CAS) is reduced by at least 2 points. 229. The method of any of embodiments 220-224, wherein the subject's Clinical Activity Score (CAS) is reduced to (1). 230. The method of any of embodiments 220-224, wherein the subject's Clinical Activity Score (CAS) is reduced to zero (0). 231. A method for treating thyroid-associated eye disease (TAO) or reducing the severity of TAO in a subject, comprising administering to the subject an antibody of any one of embodiments 1 to 3, or a pharmaceutical composition comprising the antibody, wherein treatment with the antibody (i) reduces exophthalmos in the eye by at least 2 mm; (ii) is not accompanied by a worsening of the other (i.e., fellow) eye by 2 mm or more; and (iii) reduces CAS in the subject to (1) or zero (0). 232. A method for improving quality of life in a subject suffering from thyroid-associated eye disease (TAO, also known as Graves' eye disease / Graves' ophthalmopathy), comprising administering to the subject an antibody described in any one of embodiments 211 to 213, or a pharmaceutical composition comprising said antibody. 233. The method of embodiment 232, wherein said quality of life is measured by the Graves' Eye Disease Quality of Life (GO-QoL) survey, or either its visual functioning or appearance subscale. 234. The method of embodiment 232, wherein the treatment results in an improvement of 8 or more points in GO-QoL. 235. The method described in embodiment 232, wherein the treatment results in an improvement in the functioning subscale of the GO-QoL. 236. The method of embodiment 232, wherein said treatment results in an improvement in the appearance subscale of the GO-QoL. 237. A method for treating or reducing the severity of diplopia in a subject suffering from thyroid-associated eye disease (TAO), comprising administering to the subject an antibody according to any one of embodiments 211 to 213, or a pharmaceutical composition comprising said antibody. 238. The method of embodiment 237, wherein the diplopia is stationary diplopia. 239. The method of embodiment 237, wherein the diplopia is non-stationary diplopia. 240. The method of embodiment 237, wherein the diplopia is intermittent diplopia. 241. The method of embodiment 237, wherein the improvement or reduction in the severity of diplopia persists for at least 20 weeks after discontinuing antibody administration. 242. The method of embodiment 237, wherein the improvement or reduction in the severity of diplopia persists for at least 50 weeks after discontinuing antibody administration. 243. The method of any one of embodiments 217-242, wherein the antibody is administered at a first dose of about 1 mg / kg to about 5 mg / kg, about 5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 20 mg / kg, about 20 mg / kg to about 30 mg / kg, about 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, or about 30 mg / kg of antibody. 244. The method of any one of embodiments 217-242, wherein the antibody is administered at a dosage of about 10 mg / kg to about 20 mg / kg of antibody as a first dose. 245. The method of any one of embodiments 217-242, wherein the antibody is administered at a dose of about 1 mg / kg to about 10 mg / kg, about 2 mg / kg to about 5 mg / kg, or about 5 mg / kg to about 20 mg / kg of antibody in subsequent doses. 246. The method of any one of embodiments 217-242, wherein the antibody is administered in the following amounts: about 10 mg / kg of antibody as a first dose; and about 20 mg / kg of antibody as a subsequent dose. 247. The method of embodiment 246, wherein the subsequent doses are administered every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, or every 8 weeks for at least 21 to 52 weeks, or more. 248. A method for increasing the internalization of IGF-1R in a cell, the method comprising contacting the cell with an antibody according to any one of embodiments 211 to 213 or a pharmaceutical composition comprising the antibody. 249. The method of embodiment 248, wherein said contacting comprises administering to the subject said antibody or a pharmaceutical composition comprising said antibody. 250. The method of embodiment 249, wherein the subject has or is at risk for thyroid eye disease (TED). 251. A method for inhibiting IGF-1 stimulated receptor phosphorylation in a cell, the method comprising contacting the cell with an antibody according to any one of embodiments 211 to 213, or a pharmaceutical composition comprising the antibody. 252. The method of embodiment 251, wherein the contacting comprises administering to the subject an antibody of any one of embodiments 211 to 213, or a pharmaceutical composition comprising said antibody. 253. The method of embodiment 252, wherein the subject has or is at risk for thyroid eye disease (TED). 254. The method of any one of embodiments 251-253, wherein the antibody has an IC50 of about 0.2 nm, 0.15 nm, 0.10 nm, 0.09 nm or less. 255. A method for treating thyroid eye disease in a subject, the method comprising administering to the subject an antibody of any one of embodiments 211 to 213, or a pharmaceutical composition comprising the antibody, wherein the antibody has a serum concentration in the subject of at least or about 10 μg / mL, or 20 μg / mL, 50 μg / mL, 70 μg / mL, 75 μg / mL, 80 μg / mL, 85 μg / mL, 90 μg / mL, 95 μg / mL, 100 μg / mL, or 105 μg / mL at least 1, 2, or 3 weeks after administration. 256. The method of embodiment 255, wherein the antibody or pharmaceutical composition is administered intravenously or subcutaneously.

[0232] The subject matter will now be described with reference to the following examples. These examples are provided for illustrative purposes only, and the claims should not be construed as being limited to these examples, but rather as encompassing any and all variations that become apparent as a result of the teachings provided herein. Those skilled in the art will recognize that there are a variety of non-critical parameters that could be changed or modified to yield essentially similar results. It will be easy to recognize that. [Example]

[0233] Example

[0234] Example 1: IGF-1R antibodies block IGF-1 stimulation.

[0235] Blockade of IGF-1 stimulation is measured by hyaluronan secretion in the presence of IGF-1R antibodies VRDN-2700, VRDN-03100, VRDN-02100, VRDN-02200, VRDN-02300, VRDN-02400, VRDN-02500, VRDN-01100, VRDN-02600, and VRDN-02301, all of which are disclosed herein. Immunoglobulins are purified from the serum of patients with Graves' eye disease (GO) and tested for their ability to activate TSHR and / or IGF-1R antibodies, as well as TSHR / IGF-1R crosstalk in primary cultures of GO fibroblasts. Cells are treated with M22 or GO-Ig with or without IGF-1R inhibitory antibodies, such as those provided herein, including, but not limited to, VRDN-2700, VRDN-03100, VRDN-02100, VRDN-02200, VRDN-02300, VRDN-02400, VRDN-02500, VRDN-01100, VRDN-02600, and VRDN-02301, all of which are disclosed herein. Hyaluronan (hyaluronic acid; HA) secretion is measured as the primary biological response to GO fibroblast stimulation. IGF-1R autophosphorylation is used as a direct indicator of IGF-1R activation. TSHR activation is measured by cyclic AMP (cAMP) production. The IGF-1R antibodies disclosed herein were found to effectively block HA secretion and, therefore, IGF stimulation.

[0236] Example 2: Treatment of patients with thyroid eye disease and clinical evaluation of IGF-1R antibodies in thyroid eye disease.

[0237] Subjects are provided with a fusion of an IGF-1R inhibitor antibody, such as those provided herein, including, but not limited to, VRDN-2700, VRDN-03100, VRDN-02100, VRDN-02200, VRDN-02300, VRDN-02400, VRDN-02500, VRDN-01100, VRDN-02600, and VRDN-02301, all of which are disclosed herein. The number of fusions is individualized for each subject and based on the investigator's clinical judgment. The Day 1 visit occurs within 14 days of the last visit in the previous trial. The visit window is ±1 day for weeks 1 and 4, and ±3 days for weeks 3, 6, 9, 12, 15, 18, 21, and 24. The follow-up period is for only subjects who were exophthalmos non-responders in the previous trial; subjects who relapsed in the previous trial did not participate in the follow-up period. The visit window during the follow-up period is ±7 days.

[0238] The treatment period is 24 weeks (6 months), during which teprotumumab will be administered by infusion eight times.

[0239] Subjects who are exophthalmos non-responders will participate in a 6-month follow-up period in this extension study; subjects who relapse in the lead-in study and are re-treated in this extension study will not participate in a follow-up period.

[0240] Efficacy assessments will be performed on both eyes at each assessment time point. The "study eye" (i.e., the more severely affected eye) will remain the same as that identified at the baseline (Day 1) visit of the previous study. Both eyes will be evaluated for efficacy, but the study eye will be used to assess the primary outcome. Evaluate the Tocam scale.

[0241] Efficacy will be assessed by exophthalmos (measured as an exophthalmos rating on a clinical scale of severity using a HERETER device for consistency in measurement), CAS (7-item scale), diplopia (measured as part of a clinical scale of severity), and a clinical scale of severity (including a mobility limitation rating).

[0242] Quality of life will be assessed using the GO-QoL questionnaire.

[0243] Safety will be assessed by AE and concomitant medication monitoring, immunogenicity testing, physical and ophthalmologic examinations, vital signs, clinical safety laboratory assessments (complete blood count, chemistries (including thyroid panel and HbA1C), and urinalysis), pregnancy testing (if applicable), and electrocardiogram (ECG). The study will also be monitored by a Data Safety Monitoring Board (DSMB).

[0244] Exophthalmos assessments will be performed using a Hereter exophthalmometer for consistency in measurements, and the same Hereter instrument and the same observer will be used for each assessment throughout the study (unless strictly unavoidable). Furthermore, the same intercanthal distance (ICD) will be used in each case.

[0245] Exophthalmos will be measured in each eye on Day 1 and at Weeks 6, 12, 18, and 24 (or early withdrawal (PW)) during the treatment period, and at Months 7, 9, and 12 (or PW) during the follow-up period. Measurements will be recorded under Exophthalmos on the Clinical Measures of Severity eCRF.

[0246] The antibodies are also found to be effective in treating thyroid eye disease and improving quality of life as provided herein.

[0247] Example 3: Antibodies with increased pK

[0248] Cynomolgus monkeys were administered an antibody containing the CDR of VRDN-2700 with a YTE mutation in the Fc domain at 10 mg / kg by either intravenous or subcutaneous routes, and samples were collected at 0.5, 2, and 8 hours, as well as on days 1, 3, 7, 10, 14, 21, and 28, for PK analysis by ELISA. Teprotumumab was also administered at 10 mg / kg IV as a comparator. The results, shown in Figure 1, indicate that the antibody had significantly improved PK compared to teprotumumab.

[0249] These results indicate that antibodies containing the CDRs of VRDN-2700 are likely to be administered at lower doses than teprotumumab, even when administered subcutaneously. These results were previously unexpected.

[0250] Example 4:

[0251] VRDN-1100 is an antagonist antibody against the insulin-like growth factor receptor (IGF-1R) in development for the treatment of thyroid eye disease (TED). TED is driven by thyroid-stimulating hormone receptor (TSHR) agonist autoantibodies and crosstalk between TSHR and IGF-1R. TED is characterized by the recruitment of IGF-1R- and TSHR-expressing fibrocytes within the orbital tissue, where they mediate hyaluronan deposition and orbital muscle and lipid swelling (1). IGF-1R antagonism prevents this orbital tissue swelling. It has been found to reverse and potently reduce symptoms in TED patients (2).

[0252] VRDN-1100 is a humanized monoclonal antibody that targets IGF-1R. The IGF-1R binding and antagonist properties of VRDN-1100 were analyzed.

[0253] method

[0254] Surface plasmon resonance (SPR): Antibodies were captured by immobilized anti-Fc and analyte was flowed over recombinant IGF-1R extracellular domain (ECD). Association and dissociation rate constants (ka and kd, respectively), as well as the equilibrium dissociation constant (KD), were derived by globally fitting the data to a single-site model.

[0255] Epitope binning: VRDN-1100 was immobilized on the chip surface by amine coupling and used to capture IGF-1R-ECD, after which teprotumumab was flowed over the chip.

[0256] Cell Binding: A549 human lung adenocarcinoma cells or primary human ocular choroidal fibroblasts (HOCF) were incubated with various concentrations of VRDN-1100 or teprotumumab. A single dose (50 nM) of IgG1 isotype control was used as a negative control. Unbound antibody was removed by washing, and cells were incubated with Alexa Fluor 488-goat anti-human antibody and a cell-impermeable dye to gate live cells. The median fluorescence intensity (MFI) of live cells was measured by flow cytometry, and data were analyzed using FlowJo software. Dose curves were fitted using a nonlinear regression model: log(agonist) vs. response—variable slope (four parameters).

[0257] Internalization: Cells were incubated with various concentrations of the antibody of interest for 60 minutes at 4°C and 37°C. Cells were then incubated with a 3-fold diluted FITC-labeled goat anti-human Fc secondary antibody for 30 minutes at 4°C. The MFI of live cells was measured by flow cytometry, and the data was analyzed using FlowJo software. The median fluorescence intensity (MFI) of live cells was measured by flow cytometry, and the data was analyzed using FlowJo software.

[0258] Cell surface marker expression: HOCF cells were incubated with directly labeled antibodies or IgG isotype controls at 10 μg / mL. Median fluorescence intensity (MFI) was measured by flow cytometry and data were analyzed using FlowJo software.

[0259] Antagonism: Serum-starved A549 or HOCF cells were preincubated with various concentrations of test antibody for 1 hour at 37°C, followed by stimulation with 100 ng / mL (A549) or 200 ng / mL (HOCF) IGF-1 for 7 minutes at 37°C. Phosphorylated IGF-1R (pIGF1R) in biological replicates was measured using an R&D Systems pIGF-1R ELISA according to the manufacturer's protocol, and pIGF-1R concentrations were normalized to the lowest test antibody concentration. Dose curves were fitted using a nonlinear regression model: log(inhibitor) vs. response with variable slope (four parameters).

[0260] result

[0261] VRDN-1100 binds to IGF-1R with subnanomolar affinity. Panel A of Figure 2 shows that increasing concentrations of IGF-1R-ECD bound to anti-Fc-captured VRDN-1100 or teprotumumab reveal a stepwise increase in SPR signal, supporting the binding model. This demonstrates that a global fit is possible. After washout of IGF-1R, VRDN-1100 exhibits a more sustained binding interaction. Panel B of Figure 2 shows IGF-1R-ECD strongly bound to immobilized VRDN-1100. Teprotumumab showed no binding to the IGF-1R:VRDN-1100 complex, indicating that teprotumumab and VRDN-1100 have overlapping epitopes. The data are also presented in the table shown in Figure 2.

[0262] VRDN-1100 binds with high affinity to IGF-1R on A549 cells. As shown in Figure 3, the binding of VRDN-1100 to A549 cells was investigated by flow cytometry and found to have a similar binding distribution to teprotumumab at three different concentrations. As also shown in Figure 3, the binding dose-response curve showed a VRDN-1100 EC50 of 0.1 nM. As shown in Figure 3, VRDN-1100, VRDN-2700 with M252Y, S254T, and T256E mutations in the Fc domain, and teprotumumab exhibit comparable binding at temperatures that block IGF-1R receptor internalization. Panel D shows that VRDN-1100, VRDN-2700 with M252Y, S254T, and T256E mutations in the Fc domain, and teprotumumab cause substantial levels of internalization (approximately 50%) as measured by a reduction in membrane IGF-1R receptor concentration at 37° C. and 4° C. In the bar graph in Figure 3, the leftmost bar is the isotype control, the second set of bars from the left is teprotumumab, the second set of bars from the right is VRDN-1100, and the rightmost set of bars is VRDN-2700.

[0263] HOCF as an in vitro model for TED pathology.

[0264] CD34+, Thy-1+ orbital fibroblasts have been implicated in extracellular matrix deposition and pathogenic fibrosis in TED (5). HOFCs were shown to express (Panel A) IGF-1R and (Panel B) TSHR, as well as (Panel C) CD34 and Thy-1, indicating that they can be used as an in vitro model system for IGF-1R function in TED.

[0265] VRDN-1100 binds with high affinity to IGF-1R on HOCF cells.

[0266] Figure 5 shows VRDN-1100 binding to HOCF cells, which was examined by flow cytometry and found to have similar binding to teprotumumab at three different concentrations. The bottom right corner panel of Figure 5 shows the binding dose-response curve, which showed VRDN-1100 with an EC50 of 0.4 nM.

[0267] VRDN-1100 is a subnanomolar IGF-1R antagonist. VRDN-1100 potently inhibited IGF-1-stimulated receptor phosphorylation in A549 cells (IC50 = 0.09 nM) and HOCF cells (IC50 = 0.09 nM), as shown in Figure 6, panels A and B.

[0268] These results demonstrate that the VRDN-1100 and teprotumumab epitopes on IGF-1R overlap, that VRDN-1100 binds to IGF-1R on cells with a subnanomolar EC50, that VRDN-1100 promotes IGF-1R internalization, and that VRDN-1100 inhibits IGF-1R phosphorylation with a subnanomolar IC50. Thus, VRDN-1100 binds to, antagonizes, and internalizes IGF-1R at subnanomolar concentrations, suggesting that VRDN-1100 should be used for potential and potent inhibition of the pathophysiology that drives TED.

[0269] Example 4. VRDN-2700, which has M252Y, S254T, and T256E mutations in the Fc domain, is a novel anti-IGF-1R antibody incorporating half-life extending modifications in its Fc region, as described herein, and can be used to treat thyroid eye disease (TED). The pharmacokinetic (PK) parameters of VRDN-2700 containing these Fc mutations were measured in cynomolgus monkeys relative to the marketed IGF-1R antibody, teprotumumab, and a PK model was constructed for potential human dosing regimen projects.

[0270] TED is an autoimmune condition most commonly associated with Graves' disease and hyperthyroidism, but may also be found in euthyroid or hypothyroid patients. Ophthalmopathy in TED is driven by thyroid-stimulating hormone receptor (TSHR) agonist autoantibodies and crosstalk between TSHR and IGF-1R. Pathological remodeling of the orbital and periorbital tissues results in a variety of symptoms, which may include dry eye, increased lacrimation, local inflammation, eyelid retraction, and eventual exophthalmos, diplopia, and optic nerve compression, leading to subsequent blindness.

[0271] The underlying pathology of TED is primarily the activation of the inflammatory cascade in the orbit through the recruitment of fibrocytes and immune cells. Overexpression of IGF-1R has been demonstrated in the orbit of TED patients, and it has been speculated that IGF-1R-blocking antibodies may disrupt the crosstalk between IGF-1R and TSHR, impairing the inflammatory cascade. Indeed, IGF-1R antagonism has been shown to dramatically alleviate most of the inflammatory symptoms affecting TED patients.

[0272] VRDN-2700 is a monoclonal antibody that inhibits IGF-1-mediated signaling through the IGF-1R with subnanomolar potency and incorporates clinically validated Fc modifications (M252Y, S254T, and T256E) to extend half-life. This antibody was discovered to have a more favorable PK profile than traditional IgG therapeutic antibodies, potentially offering a less burdensome treatment paradigm for patients.

[0273] VRDN-2700 with Fc mutations was administered to cynomolgus monkeys at 2, 10, and 50 mg / kg via 30-minute intravenous (IV) infusion and at 2 and 10 mg / kg via subcutaneous (SC) infusion. Teprotumumab at 10 mg / kg was also administered via 30-minute IV infusion. Serum concentrations of VRDN-2700 and teprotumumab were measured using a human IgG-specific ELISA assay. Data were analyzed using the WinNonlin noncompartmental model. A semi-mechanistic model incorporating target-mediated drug disposition was constructed using available human and cynomolgus monkey data. The data are presented below.

[0274] The table and graph in Figure 7 show a more favorable PK profile.

[0275] The table shows PK parameters ± SD. Evidence of target-mediated drug disposition (TMDD) was observed at 2 mg / kg but not at doses of 10 and 50 mg / kg, consistent with teprotumumab and other IGF-1R antibodies that reported saturation of TMDD at higher doses.

[0276] Modifications that extend the half-life of VRDN-2700 extend exposure

[0277] At equivalent doses, SC-administered VRDN-2700 with the YTE mutation achieves greater exposure than intravenously infused teprotumumab, achieving approximately twice the half-life in NHPs with an estimated bioavailability (F) of 62% for VRDN-2700 compared to SC administration using the pre-discovery formulation. Parameters are estimated ±SD and are shown in Figure 8.

[0278] The model predicts that administering VRDN-2700 at 10 mg / kg every 3 weeks or 20 mg / kg every 6 weeks will result in a Cmin greater than 100 μg / mL, similar to the approved teprotumumab regimen (10 mg / kg first dose, followed by seven 20 mg / kg doses q3w). The 10 mg / kg q3w regimen results in a lower Cmax. A longer dosing interval would increase patient convenience and reduce treatment costs, while a lower dose and lower Cmax may potentially mitigate toxicity. Furthermore, the model predicts that a fixed dose of 300 mg subcutaneously administered weekly VRDN-2700 may achieve a steady-state Cmin of approximately 130 μg / mL, allowing for self-administration at home. If lower Cmin values ​​are beneficial, a fixed dose of 300 mg administered subcutaneously every other week of VRDN-2700 is expected to achieve steady-state Cmin levels of approximately 50 μg / mL. In summary, the extended half-life of VRDN-2700 is expected to offer patients a wider range of options for more convenient dosing intervals and routes of administration.

[0279] Example 5: Properties of VRDN2700 During antibody investigation, the expression of VRDN-2700 was compared with other antibodies with mutations in the Fc domain, such as the L / S mutations described herein. Unexpectedly, the yield for the antibody with the YTE mutation in the Fc domain (VRDN2700) was approximately 80% higher than the yield for a similar antibody without the L / S mutation. This was surprising and unexpected, as other antibodies tested targeting IGF-1R with YTE or LS mutations had similar expression regardless of Fc mutation. The YTE version had fewer low molecular weight species compared to the LS version. Thus, the YTE antibody had fewer impurities, indicating a more homogeneous composition, which provides an advantage over antibodies with LS mutations. This was also unexpected, as other antibodies investigated showed the opposite effect on such species. Furthermore, during purification, it was found that the LS mutant formed more aggregates when purified on a cation exchange column compared to VRDN-2700. Aggregation of the LS variants poses significant manufacturing challenges that were not observed with VRDN-2700. Thus, this difference in the Fc variants for this antibody could not have been anticipated or predicted, and provides a significant and unexpected advantage to the antibody referred to herein as VRDN-2700.

[0280] The extended half-life of VRDN-2700(YTE) indicates that it can be used as a convenient SC injection or as an IV infusion, requiring less therapeutic doses and / or less frequent treatments than conventional therapeutic IgG antibodies, and has superior properties compared to other Fc variant versions of the same antibody (same variable region).

[0281] Example 6: VRDN-1100 with YTE or YTE / C22S mutations binds to IGF-1R and inhibits IGF-1R autophosphorylation. The binding of VRDN-1100 with the Fc YTE mutation (SEQ ID NO: 94) in its heavy chain or the C22S and Fc YTE mutations (SEQ ID NO: 95) in its heavy chain to IGF-1R was investigated in a cell-based binding assay (A549 cells). The light chain has the sequence of SEQ ID NO: 93. The YTE Fc mutant version of VRDN1100 was found to bind to A549 cells with an EC50 of 0.30 nM, while the C22S and Fc YTE mutants had an EC50 of 0.36 nM. The ability of the antibodies to inhibit IGF-1R autophosphorylation was also investigated. The YTE mutant alone had an IC50 of 0.40 nM, while the C22S and YTE mutants had an IC50 of 0.37 nM. Thus, the antibody was found to be capable of both binding to IGF-1R and inhibiting its autophosphorylation.

[0282] Example 7: VRDN-1100 with a C22S mutation binds to IGF-1R. VRDN-1100 with a C22S mutation (SEQ ID NO: 96) in the heavy chain and , a variant of VRDN-1100 having a VL comprising the sequence of SEQ ID NO: 97 was investigated for binding to IGF-1R. Using this assay, at a pH of 7.4, the antibody exhibited a 1.04 x 10 5 k a (1 / Ms), 2.18×10 -5 k d (1 / s) and 2.10×10 -10 K D (M) was found to bind to IGF-1R.

[0283] Each of these examples, and the embodiments provided herein, demonstrate that the antibodies provided herein can be used to treat TED and its associated conditions.

[0284] All references cited herein are incorporated by reference to the same extent as if each individual publication, database entry (e.g., GenBank sequence or GeneID entry), patent application, or patent were specifically and individually indicated and incorporated by reference. This statement of incorporation by reference is intended by applicants to relate to each and every individual publication, database entry (e.g., GenBank sequence or GeneID entry), patent application, or patent, each of which is expressly identified in compliance with 37 CFR §1.57(b)(1), even if such citation is not immediately adjacent to the specific statement of incorporation by reference, in compliance with 37 CFR §1.57(b)(2). The inclusion within this specification of a specific statement of incorporation by reference, if any, does not in any way weaken this general statement of incorporation by reference. The citation of any reference herein is not intended as an admission that the reference cites pertinent prior art, nor is it an admission as to the contents or date of such publication or document.

[0285] The present embodiments are not limited in scope by the specific embodiments described herein. Indeed, various modifications in addition to those described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are intended to be included within the scope of the embodiments and any accompanying claims.

[0286] The specification is considered to be sufficient to enable one skilled in the art to practice the embodiments. Various modifications, in addition to those shown and described herein, will become apparent to those skilled in the art from the foregoing description and will fall within the scope of the present disclosure and any appended claims.

Claims

1. A pharmaceutical composition comprising an antibody for treating thyroid eye disease in a subject, wherein the antibody comprises a light chain variable region having the amino acid sequence of SEQ ID NO: 13 and a heavy chain variable region having the amino acid sequence of SEQ ID NO:

14.

2. The pharmaceutical composition of claim 1 , wherein the antibody comprises an Fc region with M428L and N434S substitutions.

3. 2. The pharmaceutical composition of claim 1, wherein the antibody comprises an Fc region with M428L, N434S, M252Y, S254T, and T256E substitutions.

4. 2. The pharmaceutical composition of claim 1, wherein the antibody comprises an Fc region with M252Y, S254T, and T256E substitutions.

5. 2. The pharmaceutical composition of claim 1, wherein the antibody comprises a light chain having the amino acid sequence of SEQ ID NO:93 and a heavy chain having the amino acid sequence of SEQ ID NO:

92.

6. 2. The pharmaceutical composition of claim 1, wherein the antibody comprises a light chain having the amino acid sequence of SEQ ID NO:93 and a heavy chain having the amino acid sequence of SEQ ID NO:

94.

7. 2. The pharmaceutical composition of claim 1, wherein the antibody comprises a light chain having the amino acid sequence of SEQ ID NO:93 and a heavy chain having the amino acid sequence of SEQ ID NO:

95.

8. 10. A method for treating thyroid eye disease in a subject, said method comprising administering a pharmaceutical composition comprising the antibody of any one of claims 1 to 7.

9. 9. The method of claim 8, wherein the pharmaceutical composition is administered intravenously.

10. 9. The method of claim 8, wherein the pharmaceutical composition is administered subcutaneously.

11. 10. A method for treating thyroid-associated eye disease (TAO), or a symptom of TAO, or reducing the severity of TAO, or a symptom of TAO, said method comprising administering to a subject a pharmaceutical composition according to any one of claims 1 to 7.

12. 10. A method of reducing exophthalmos in a subject suffering from thyroid-associated eye disease (TAO), said method comprising administering to the subject a pharmaceutical composition according to any one of claims 1 to 7.

13. 10. A method of treating thyroid eye disease in a subject, said method comprising administering the pharmaceutical composition of any one of claims 2 to 4.

14. 10. A method for reducing Clinical Activity Score (CAS) of Thyroid Associated Eye Disease (TAO) in a subject, the method comprising administering to the subject the pharmaceutical composition of any one of claims 1 to 7.

15. 10. A method of a) reducing exophthalmos by at least 2 mm and b) lowering Clinical Activity Score (CAS) in a subject suffering from thyroid-associated eye disease (TAO), said method comprising administering to the subject a pharmaceutical composition according to any one of claims 1 to 7. method.

16. 16. The method of any one of claims 11 to 15, wherein exophthalmos is reduced by at least 2 mm.

17. 16. The method of any one of claims 11 to 15, wherein exophthalmos is reduced by at least 3 mm.

18. 16. The method of any one of claims 11 to 15, wherein exophthalmos is reduced by at least 4 mm.

19. 16. The method of any one of claims 11 to 15, wherein the subject's Clinical Activity Score (CAS) is reduced by at least 2 points.

20. The method of any one of claims 11 to 15, wherein the subject's Clinical Activity Score (CAS) is reduced to (1).

21. 16. The method of any one of claims 11 to 15, wherein the subject's Clinical Activity Score (CAS) is reduced to zero (0).

22. 10. A method of treating or reducing the severity of thyroid-associated eye disease (TAO) in a subject, comprising administering to the subject the pharmaceutical composition of any one of claims 1 to 7, wherein treatment with the antibody (i) reduces exophthalmos in one eye by at least 2 mm; (ii) is not accompanied by a worsening of the other (i.e., fellow) eye by 2 mm or more; and (iii) reduces CAS in the subject to (1) or zero (0).

23. 10. A method for improving quality of life in a subject suffering from thyroid-associated eye disease (TAO, also known as Graves' eye disease / Graves' ophthalmopathy), the method comprising administering to the subject a pharmaceutical composition according to any one of claims 1 to 7.

24. 24. The method of claim 23, wherein the quality of life is measured by the Graves' Eye Disease Quality of Life (GO-QoL) survey or either the visual functioning or appearance subscales thereof.

25. 24. The method of claim 23, wherein the treatment results in an improvement of 8 or more points in GO-QoL.

26. 24. The method of claim 23, wherein the treatment results in an improvement in the functioning subscale of the GO-QoL.

27. 24. The method of claim 23, wherein the treatment results in an improvement in the appearance subscale of the GO-QoL.

28. 10. A method for treating or reducing the severity of diplopia in a subject suffering from thyroid-associated eye disease (TAO), said method comprising administering to the subject a pharmaceutical composition according to any one of claims 1 to 7.

29. 29. The method of claim 28, wherein the diplopia is stationary diplopia.

30. 29. The method of claim 28, wherein the diplopia is non-stationary diplopia.

31. 29. The method of claim 28, wherein the diplopia is intermittent diplopia.

32. 29. The method of claim 28, wherein the improvement or reduction in the severity of diplopia persists for at least 20 weeks after administration of the antibody is discontinued.

33. 29. The method of claim 28, wherein the improvement or reduction in the severity of diplopia persists for at least 50 weeks after administration of the antibody is discontinued.

34. 34. The method of any one of claims 8-33, wherein the pharmaceutical composition is administered at a dosage of about 1 mg / kg to about 5 mg / kg, about 5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 20 mg / kg, about 20 mg / kg to about 30 mg / kg, about 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, or about 30 mg / kg of antibody as a first dose.

35. 34. The method of any one of claims 8 to 33, wherein the pharmaceutical composition is administered at a dosage of about 10 mg / kg to about 20 mg / kg of antibody as a first dose.

36. 34. The method of any one of claims 8-33, wherein the pharmaceutical composition is administered at a dosage of about 1 mg / kg to about 10 mg / kg, about 2 mg / kg to about 5 mg / kg, or about 5 mg / kg to about 20 mg / kg of antibody, with subsequent doses.

37. 34. The method of any one of claims 8 to 33, wherein the pharmaceutical composition is administered in the following amounts: about 10 mg / kg of antibody as a first dose; and about 20 mg / kg of antibody as subsequent doses.

38. 38. The method of claim 37, wherein the subsequent doses are administered every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, or every 8 weeks for at least 21 to 52 weeks, or more.

39. 8. A method for increasing the internalization of IGF-1R in a cell, said method comprising contacting said cell with the pharmaceutical composition of any one of claims 1 to 7.

40. 40. The method of claim 39, wherein said contacting comprises administering to the subject the pharmaceutical composition of any one of claims 1 to 7.

41. 41. The method of claim 40, wherein the subject has or is at risk for thyroid eye disease (TED).

42. 8. A method for inhibiting IGF-1 stimulated receptor phosphorylation in a cell, said method comprising contacting said cell with the pharmaceutical composition of any one of claims 1 to 7.

43. 43. The method of claim 42, wherein said contacting comprises administering to the subject the pharmaceutical composition of any one of claims 1 to 7.

44. 44. The method of claim 43, wherein the subject has or is at risk for thyroid eye disease (TED).

45. 45. The method of any one of claims 42-44, wherein the antibody has an IC50 of about 0.2 nm, 0.15 nm, 0.10 nm, 0.09 nm or less.

46. 10. A method of treating thyroid eye disease in a subject, the method comprising administering to the subject the pharmaceutical composition of any one of claims 1-7, wherein the antibody has a serum concentration in the subject of at least or about 10 μg / mL, or 20 μg / mL, 50 μg / mL, 70 μg / mL, 75 μg / mL, 80 μg / mL, 85 μg / mL, 90 μg / mL, 95 μg / mL, 100 μg / mL, or 105 μg / mL at least 1, 2, or 3 weeks after administration.

47. 47. The method of claim 46, wherein the pharmaceutical composition is administered intravenously or subcutaneously.

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