Compositions, Dosages and Methods for Treating Thyroid Eye Disease - Patent application

Abstract

Provided herein are antibodies and compositions against IGF-1R and their uses.The IGF-1R antibodies and antigen-binding fragments inhibit IGF-1R function or block the biological function of IGF-I-mediated IGF-1R signal transduction.The present invention relates to the use of IGF-1R antibodies and antigen-binding fragments to treat thyroid eye disease (TAO), also known as thyroid eye disease (TED), Graves' ophthalmopathy or orbitopathy (GO), thyrotoxic exophthalmopathy, thyroid dystrophic ophthalmopathy, and other thyroid eye disorders related to IGF-1R signal transduction.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 260,133, filed August 10, 2021, and U.S. Provisional Application No. 63 / 261,744, filed September 28, 2021, each of which is incorporated by reference in its entirety herein.

[0002] Reference to an Electronically Submitted Sequence Listing This application contains a Sequence Listing that has been submitted electronically in XML file format, which is hereby incorporated by reference in its entirety. The XML copy, created on August 10, 2022, is named "257635_000402_ST26.xml" and is 12,855 bytes in size. [Background technology]

[0003] background Thyroid eye disease (TAO), also known as thyroid eye disease (TED), Graves' ophthalmopathy or orbitopathy (GO), thyrotoxic exophthalmos, thyrotoxic ophthalmopathy, and several other terms, is an orbital disease associated with thyroid dysfunction. TAO is divided into two types. Active TAO, which typically lasts 1-3 years, is characterized by a continuing autoimmune / inflammatory response in the soft tissues of the orbit. Active TAO causes expansion and remodeling of the soft tissues of the eye. The autoimmune / inflammatory response of active TAO resolves spontaneously, and the condition transitions to inactive TAO. Inactive TAO is the term used to describe the long-term / permanent sequelae of active TAO. The cause of TAO is unknown. TAO is typically associated with Graves' hyperthyroidism, but may also occur as part of other autoimmune conditions that affect the thyroid gland and produce orbital and periorbital tissue pathology and rarely pretibial skin pathology (pretibial myxedema) or digital pathology (thyroid clubbing). TAO is an autoimmune orbital disease that primarily affects the orbit and periorbital soft tissues with secondary effects on the eyes and vision. In TAO, the eyes are pushed forward (distended) from their sockets as a result of inflammation and enlargement of the orbital soft tissues, primarily the eye muscles and fat. This is a phenomenon called proptosis or exophthalmos. Although most cases of TAO do not result in vision loss, the condition can cause sight-threatening lagophthalmos, bothersome diplopia (double vision), and compressive thyroid optic neuropathy. TAO may precede, occur simultaneously with, or occur after the systemic complications of hypothyroidism. Ocular manifestations of TAO include upper eyelid retraction, eyelid lag, swelling, redness (erythema), conjunctivitis, and eye globe enlargement (exophthalmos or proptosis), conjunctival edema, periorbital edema, and altered ocular motility with significant functional, social, and cosmetic consequences. Many of the signs and symptoms of TAO, including exophthalmos and ocular hyperemia, result from the expansion of orbital adipose tissue and periorbital muscles. Adipose tissue volume increases in part due to new fat cell development (adipogenesis) within the orbital fat. Accumulation of hydrophilic glycosaminoglycans, primarily hyaluronic acid, within the orbital adipose tissue and within the perimysial connective tissue between extraocular muscle fibers further expands the adipose compartment and increases the extraocular muscle bodies.Hyaluronan is produced by fibroblasts present in ocular fat and extraocular muscles, and its synthesis in vitro is stimulated by several cytokines and growth factors, including IL-1 beta, interferon-gamma, platelet-derived growth factor, thyroid-stimulating hormone (TSH), and insulin-like growth factor I (IGF-I).

[0004] Antibodies that activate the insulin-like growth factor I receptor (IGF-IR) have also been detected and are 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 blockade of IGF-IR appears to attenuate both IGF-1- and TSH-dependent signaling. It has been proposed that blockade of IGF-IR using antibody antagonists may reduce both TSHR- and IGF-I-dependent signaling, thus preventing the pathological activity of autoantibodies that act as agonists against either receptor.

[0005] IGF-IR is a widely expressed heterotetrameric protein involved in regulating the growth and metabolic functions of many cell types. It is a tyrosine kinase receptor that contains two subunits. IGF-IR alpha contains the ligand binding domain, while IGF-IR beta is involved in signal transduction and contains the tyrosine phosphorylation site.

[0006] Current treatments for hyperthyroidism due to Graves' disease are incomplete because there are no treatments that target the specific underlying pathogenic autoimmune mechanisms of the disease. The treatment of moderate to severe active TAO is much more complex. Although there has been a better understanding of its pathogenesis in recent years, TAO still presents therapeutic challenges and dilemmas. There are no approved drugs to treat active TAO. Intravenous glucocorticoids (ivGC) and oral glucocorticoids have been used to treat patients with moderate to severe active TAO, but the results are rarely satisfactory. Partial responses are common, and relapse (rebound) after withdrawal is not uncommon. Adverse events occur, and many patients eventually require functional restoration surgery, which is performed when their condition transitions to inactive TAO. Thus, there is still a need to provide alternative therapies for TAO and its associated symptoms. Summary of the Invention [Means for solving the problem]

[0007] Abstract The present disclosure generally relates to IGF-1R antibodies, and their antigen-binding fragments and their uses.Certain IGF-1R antibodies and antigen-binding fragments inhibit IGF-1R function or block the biological function of IGF-I-mediated IGF-1R signal transduction.In addition, the present invention generally relates to a method for treating thyroid eye disease (TAO), also known as thyroid eye disease (TED), Graves' ophthalmopathy or orbitopathy (GO), thyrotoxic exophthalmos, thyroid dystrophic ophthalmopathy, and other thyroid eye disorders related to IGF-1R signal transduction.

[0008] In some embodiments, the disclosure provides a method of treatment that includes treating thyroid eye disease in a subject in need of such treatment, comprising administering a first dose of an antibody intravenously or subcutaneously to the subject, wherein the first dose is selected from the group consisting of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg. and administering intravenously or subcutaneously to the subject one or more subsequent doses of the antibody, each subsequent dose being selected from the group consisting of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg, wherein the antibody is as defined herein.

[0009] In some embodiments, the disclosure provides a method of treating thyroid eye disease in a subject in need of such treatment, comprising administering a first dose of an antibody intravenously or subcutaneously to the subject, the first dose being selected from the group consisting of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg, and administering one or more subsequent doses of the antibody intravenously or subcutaneously to the subject. and wherein each subsequent dose is selected from the group consisting of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg, wherein the antibody is as defined herein, and the one or more subsequent doses are administered when the subject has not responded adequately to the one or more previous doses, as determined by clinical activity scores and / or exophthalmos measurements.

[0010] In some embodiments, the disclosure provides a first dose of about 2 mg / kg, about 3 mg / kg, about 2.5 mg / kg, about 5 mg / kg, about 7.5 mg / kg, about 10 mg / kg, about 15 mg / kg, or about 20 mg / kg.

[0011] In some embodiments, at least one of the one or more subsequent doses is about 2 mg / kg, about 3 mg / kg, about 5 mg / kg, about 7.5 mg / kg, about 10 mg / kg, about 15 mg / kg, or about 20 mg / kg.

[0012] In some embodiments, the first dose is about 10 mg / kg. In some embodiments, the one or more subsequent doses are about 10 mg / kg.

[0013] In some embodiments, the disclosure provides methods further comprising administering to the subject one or more loading doses of the antibody prior to administering the first dose.

[0014] In some embodiments, the disclosure provides a method, further comprising administering to the subject a first loading dose of the antibody prior to administering the first dose, wherein the first loading dose is selected from the group consisting of about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 12.5 mg / kg, or about 12.5 mg / kg to about 15 mg / kg.

[0015] In some embodiments, the first loading dose is about 5 mg / kg, 7.5 mg / kg, 10 mg / kg, 12.5 mg / kg, 15 mg / kg, or 20 mg / kg.

[0016] In some embodiments, the second loading dose is about 5 mg / kg, 7.5 mg / kg, 10 mg / kg, 12.5 mg / kg, 15 mg / kg, or 20 mg / kg.

[0017] In some embodiments, the disclosure provides a method of improving treatment of thyroid eye disease in a subject who has previously received one or more treatments, comprising administering to the subject intravenously or subcutaneously at least one dose of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg of an antibody, wherein the antibody is an antibody as defined herein, e.g., the antibody comprises a heavy chain and a light chain, and the heavy chain has the sequence The antibody comprises an HCDR1 of SEQ ID NO: 7, an HCDR2 of SEQ ID NO: 8, and an HCDR3 of SEQ ID NO: 9, and a light chain comprises an LCDR1 of SEQ ID NO: 4, an LCDR2 of SEQ ID NO: 5, and an LCDR3 of SEQ ID NO: 6; or the antibody comprises a light chain comprising a variable region having the amino acid sequence of SEQ ID NO: 2, and the heavy chain comprises a variable region sequence having the amino acid sequence of SEQ ID NO: 3; or the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 11, and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10; wherein at least one dose results in an improvement in one or more measured values ​​compared to the one or more measured values ​​prior to the at least one dose.

[0018] In some embodiments, the one or more measurements are selected from exophthalmos, CAS, level of deterioration in the fellow eye, score on GO-QoL, and combinations thereof.

[0019] In some embodiments, if the subject does not exhibit a satisfactory response after at least one dose, one or more subsequent doses of the antibody are administered to the subject, each dose being selected from the group consisting of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg.

[0020] In some embodiments, the one or more subsequent doses improve one or more of: exophthalmos, CAS, level of deterioration in the fellow eye, score on GO-QoL, and combinations thereof, compared to before the one or more subsequent doses.

[0021] The present disclosure provides a method of treating thyroid eye disease in a subject in need thereof, comprising the steps of administering a first dose of an antibody intravenously or subcutaneously to the subject, wherein the first dose is selected from the group consisting of about 250 mg, about 300 mg, about 350 mg, or about 400 mg, and administering one or more subsequent doses of the antibody intravenously or subcutaneously to the subject, wherein each subsequent dose is selected from the group consisting of about 250 mg, about 300 mg, about 350 mg, or about 400 mg, wherein the antibody is as defined herein.

[0022] As used herein, an antibody is defined herein and may include a heavy chain and a light chain. In some embodiments, the antibody includes a light chain comprising the amino acid sequence of SEQ ID NO: 11, and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10. In some embodiments, the heavy chain may include Fc mutations in the Fc domain, such as M252Y, S254T, and T256E mutations. In some embodiments, the heavy chain includes a VH having an amino acid sequence of SEQ ID NO: 3. In some embodiments, the light chain includes a VL comprising the amino acid sequence of SEQ ID NO: 2. Thus, in some embodiments, the antibody includes a VL of SEQ ID NO: 2, and a VH of SEQ ID NO: 3. In some embodiments, the antibody includes a heavy chain variable region (VH) comprising an HCDR1 of SEQ ID NO: 7, an HCDR2 of SEQ ID NO: 8, and an HCDR3 of SEQ ID NO: 9, and a light chain variable region (VL) comprising an LCDR1 of SEQ ID NO: 4, an LCDR2 of SEQ ID NO: 5, and an LCDR3 of SEQ ID NO: 6.

[0023] In some embodiments, the first dose is about 250 mg, 300 mg, 350 mg, or 400 mg.

[0024] In some embodiments, the one or more subsequent doses is about 250 mg, 300 mg, 350 mg, or 400 mg.

[0025] In some embodiments, the disclosure provides methods further comprising administering to the subject one or more loading doses of the antibody prior to administering the first dose.

[0026] In some embodiments, the disclosure provides a method, further comprising administering to the subject a first loading dose of the antibody prior to administering the first dose, wherein the first loading dose is selected from the group consisting of about 250 mg, 300 mg, 350 mg, or 400 mg.

[0027] In some embodiments, the first loading dose is about 250 mg, 300 mg, 350 mg, or 400 mg.

[0028] In some embodiments, the second loading dose is about 250 mg, 300 mg, 350 mg, or 400 mg.

[0029] In some embodiments, the one or more subsequent doses are the same in amount as the first dose.

[0030] In some embodiments, the amount of one or more subsequent doses is different from the amount of the first dose.

[0031] In some embodiments, at least one of the one or more subsequent doses is administered 1, 2, 3, 4, 5, 6, or 8 weeks after the first dose.

[0032] In some embodiments, only 1, 2, 3, 4, 5, 6, or 7 subsequent doses are administered to the subject.

[0033] In some embodiments, a total of 2, 3, 4, 5, 6, 7, or 8 doses are administered to the subject.

[0034] In some embodiments, after two or three doses of the antibody, the subject's clinical activity score is reduced.

[0035] In some embodiments, each subsequent dose is administered 1, 2, 3, 4, 5, 6, 7, or 8 weeks after the previous dose.

[0036] In some embodiments, at least one dose is administered by intravenous infusion over a period of from 45 minutes to about 90 minutes, or over a period of from 60 minutes to about 90 minutes.

[0037] In some embodiments, at least one dose is administered subcutaneously. In some such embodiments, the subcutaneous administration is self-administered.

[0038] In some embodiments, a second loading dose of the antibody is administered to the subject after the first loading dose, and the first and second loading doses are administered before the first dose.

[0039] In some embodiments, the first loading dose and the second loading dose are the same dose amount.

[0040] In some embodiments, the first loading dose and the second loading dose are different dose amounts.

[0041] In some embodiments, a first loading dose is administered to the subject 1, 2, 3, or 4 weeks before the first dose is administered.

[0042] In some embodiments, the antibody is administered as part of a pharma- ceutically acceptable composition comprising the antibody and at least one pharma- ceutically acceptable excipient, and the antibody has a solubility in the pharma- ceutically acceptable composition of at least about 150 mg / ml.

[0043] In some embodiments, the subject has shown unsatisfactory response to previous treatment for thyroid eye disease.In some such embodiments, unsatisfactory response is one or more of the following: cannot reduce exophthalmos by 2mm or more; cannot reduce CAS for one or more components, or by 2 points or more; worsening of other eye by 2mm or more; cannot reduce diplopia; cannot continue to improve diplopia for a period of time; cannot improve the score of Graves' Ophthalmopathy Quality of Life (GO-QoL) assessment by 8 points or more; and combinations thereof.

[0044] In some embodiments, the first loading dose and the second loading dose are administered about 1, about 2, or about 3 weeks apart.

[0045] In some embodiments, the second loading dose is administered about 1, about 2, or about 3 weeks before the first dose.

[0046] In some embodiments, a method is provided for treating thyroid eye disease in a subject in need of such treatment, comprising the step of intravenously administering a first dose of 10 mg / kg of an antibody to the subject, wherein the antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises an HCDR1 of SEQ ID NO: 7, an HCDR2 of SEQ ID NO: 8, and an HCDR3 of SEQ ID NO: 9, and the light chain comprises an LCDR1 of SEQ ID NO: 4, an LCDR2 of SEQ ID NO: 5, and an LCDR3 of SEQ ID NO: 6.

[0047] In some embodiments, the method further comprises administering a subsequent dose of about 10 mg / kg. In some embodiments, the subsequent dose is administered about 3 weeks after the first dose. In some embodiments, the method further comprises administering a subsequent dose of about 10 mg / kg every 3 weeks after the first dose. In some embodiments, the subsequent dose is administered every 3 weeks for a total of 4 subsequent doses. In some embodiments, the subsequent dose is administered every 3 weeks for a total of 7 subsequent doses. In some embodiments, the subject has a reduction in exophthalmos and an improvement in CAS score within 3 weeks or 6 weeks of the first dose. [Brief description of the drawings]

[0048] [Figure 1-1] 1A-D show various embodiments defined herein. [Figure 1-2] Same as above.

[0049] [Figure 2-1] 2A-B show various embodiments defined herein. [Figure 2-2] Same as above.

[0050] [Figure 3-1] 3A-F show various embodiments defined herein. [Figure 3-2] Same as above. [Figure 3-3] Same as above.

[0051] [Figure 4-1] 4A-C show various embodiments defined herein. [Figure 4-2] Same as above. [Figure 4-3] Same as above.

[0052] [Figure 5-1] 5A-B show various embodiments defined herein. [Figure 5-2] Same as above.

[0053] [Figure 6] 6A-B show various embodiments defined herein.

[0054] [Figure 7] FIG. 7 illustrates various embodiments defined herein.

[0055] [Figure 8] FIG. 8 illustrates various embodiments defined herein.

[0056] [Figure 9] FIG. 9 illustrates various embodiments defined herein.

[0057] [Figure 10-1] 10A-C show various embodiments defined herein. [Figure 10-2] Same as above. [Figure 10-3] Same as above.

[0058] [Figure 11] FIG. 11 illustrates various embodiments defined herein.

[0059] [Figure 12-1] 12A-C show various embodiments defined herein. [Figure 12-2] Same as above. [Figure 12-3] Same as above.

[0060] [Figure 13-1] 13A-B show various embodiments defined herein. [Figure 13-2] Same as above.

[0061] [Figure 14-1] 14A-B show various embodiments defined herein. [Figure 14-2] Same as above. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0062] Detailed Description Provided herein are antibodies that bind to and modulate the activity of IGF-1R. The antibodies can be used, for example, to treat thyroid eye disease.

[0063] As used herein, "thyroid eye disease" (TAO), "thyroid eye disease" (TED), "Graves' ophthalmopathy" or "Graves' orbitopathy" (GO) refer to the same disorder or condition and are used interchangeably. They all refer to an inflammatory orbital pathology associated with some autoimmune thyroid disorder, most commonly "Graves' disease" (GD), but sometimes with other diseases, such as Hashimoto's thyroiditis.

[0064] The terms "proptosis" and "exophthalmos" (also known as exophthalmus, exophthalmia, or exorbitism) refer to the forward projection, movement, expansion, or protrusion of an organ. As used herein, these terms refer to the forward projection, movement, expansion, or protrusion of the eye from the orbit to the front. Exophthalmos and exophthalmos are considered by some skilled in the art to have the same meaning and are often used interchangeably, while others attribute the difference to subtle differences in their meaning. Some use exophthalmos to refer to severe proptosis or to refer to endocrine-related exophthalmos. Additionally, others use the term exophthalmos to describe exophthalmos associated with the eye, for example, the eye of a subject with TAO (TED or GO).

[0065] As used herein, the terms "exophthalmos" and "exophthalmos" are used interchangeably and refer to the forward projection, movement, expansion or protrusion of the eye from the orbit to the front. Due to the hard bony structure of the orbit that only opens to the front for expansion, any increase in orbital soft tissue contents occurring from the side or back will cause the eye to move forward. Exophthalmos or exophthalmos can be the result of several disease processes, including infection, inflammation, tumor, trauma, metastasis, endocrine pathology, vascular disease and extraorbital pathology. TAO (TED or GO) is now considered the most common cause of exophthalmos in adults. Exophthalmos can be bilateral, as is common with TAO (TED or GO), or unilateral, as is common with orbital tumors.

[0066] Measurement of the degree of exophthalmos may be performed, for example, using an exophthalmometer, an instrument used to measure the degree of anterior movement of the eye. This device allows for the measurement of the anterior distance from the lateral orbital margin to the corneal apex. Computed tomography (CT) scanning and magnetic resonance imaging (MRI) may also be used to evaluate the degree of exophthalmos or proptosis. CT scanning is an excellent imaging technique for the diagnosis of TAO. In addition to allowing visualization of the enlarged extraocular muscles, CT scans provide the surgeon or clinician with a depiction of the bony anatomy of the orbit when orbital decompression is required. MRI, with its multiplanar and inherent contrast capabilities, provides excellent imaging of the orbital contents without the radiation exposure associated with CT scan studies. MRI provides better imaging of the optic nerve, orbital fat, and extraocular muscles, while CT scans provide better images of the bony structures of the orbit. Orbital ultrasonography may also be used to diagnose and evaluate TAO because it can be performed quickly and reliably. The high reflectivity and dilation of the extraocular muscles are easily assessed, and serial ultrasound examinations may also be used to assess the progression or stability of the ophthalmopathy. Based on currently available or future technology, one of skill in the art will be able to determine the best approach to diagnose and assess the degree of exophthalmos or exophthalmos.

[0067] As used herein, the term "antibody" refers to any form of antibody that exhibits the desired biological activity.The term is therefore used in the broadest sense, specifically including, but 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 that is obtained by exposure to an antigen of the immune system before modification of the antibody for intended use, for example, humanization of the antibody for use as a human therapeutic antibody.

[0068] As used herein, unless otherwise indicated, "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 and multispecific antibodies formed from antibody fragments.

[0069] A "Fab fragment" is a fragment of one light chain and one heavy chain. H 1 and the variable region of the heavy chain of a Fab molecule. The heavy chain of a Fab molecule cannot form disulfide bonds with another heavy chain molecule.

[0070] The "Fc" region is the C region of an antibody. H 1 and C H The two heavy chain fragments contain two domains. The two heavy chain fragments are connected by two or more disulfide bonds and by C H The three domains are held together by hydrophobic interactions.

[0071] In some embodiments, the antibody or antigen fragment herein comprises an Fc region. In some embodiments, the Fc region comprises a mutation that extends the half-life of the antibody when linked 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. 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, L235E, M428L, and N434S mutations. In some embodiments, the Fc region comprises M428L and N434S mutations. In some embodiments, the Fc region comprises 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 herein by reference in its entirety. The numbering referred to herein refers to the Kabat numbering system for the Fc region.

[0072] A "Fab' fragment" is a fragment that contains one light chain and one V H Domain and C H 1 domain and also C H 1 Domain and C H and a portion or fragment of one heavy chain that contains the region between the two domains, such that interchain disulfide bonds can form between the two heavy chains of the two Fab' fragments to form an F(ab')2 molecule.

[0073] "F(ab')2 fragment" is a fragment of two light chains and H 1 Domain and C H2 The F(ab')2 fragment contains two heavy chains that contain a portion of the interdomain constant region, such that an interchain disulfide bond is formed between the two heavy chains. Thus, the F(ab')2 fragment is composed of two Fab' fragments held together by disulfide bonds between the two heavy chains.

[0074] The "Fv region" comprises the variable regions from both the heavy and light chains, but lacks the constant regions.

[0075] The term "single chain Fv" or "scFv" antibody refers to the V H and V L It refers to an antibody fragment comprising scFv domains, which domains are present in a single polypeptide chain. Generally, the Fv polypeptide comprises a polypeptide linker between the V and V polypeptide chains that enables the scFv to form the desired structure for antigen binding. H Domain and V L Further included between the domains. For a review of scFvs, see 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. WO88 / 01649, and U.S. Patent Nos. 4,946,778 and 5,260,203.

[0076] A "domain antibody" is an immunologically functional immunoglobulin fragment that contains only the variable region of a heavy chain and the variable region of a light chain. In some cases, two or more V H The domains are covalently linked by a peptide linker to create a bivalent domain antibody. H The regions may target the same antigen or may target different antigens.

[0077] A "bivalent antibody" comprises 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).

[0078] In certain embodiments, the monoclonal antibodies herein also include camelized single domain antibodies. See, e.g., Muyldermans et al. (2001) Trends Biochem. Sci. 26:230; Reichmann et al. (1999) J. Immunol. Methods 231:25; WO94 / 04678; WO94 / 25591; U.S. Patent No. 6,005,079). In one embodiment, the present invention provides a method for the preparation of camelized single domain antibodies by the use of two V-domain antibodies with modifications such that single domain antibodies are formed. H The present invention provides single domain antibodies comprising the domains.

[0079] As used herein, the term "diabody" refers to a small antibody fragment with two antigen-binding sites, which contains a light chain variable domain (V L ) connected to a heavy chain variable domain (V H )(V H -V L or V L -V H ). The use of a linker that is too short to allow pairing between the two domains on the same chain forces the domains to pair with complementary domains on another chain, creating two antigen-binding sites. Diabodies are described in more detail, for example, in EP 404,097; WO 93 / 11161; and Holliger et al. (1993) Proc. Natl. Acad. Sci. USA 90: 6444-6448. For a review of engineered antibody variants, see generally Holliger and Hudson (2005) Nat. Biotechnol. 23:1126-1136.

[0080] 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 parent antibody that is modified) when the 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 contemplated that the antibodies or antigen-binding fragments of the present invention may include conservative or non-conservative amino acid substitutions, sometimes referred to as "conservative variants" or "function-conservative variants" of antibodies, whose biological activity is not substantially altered.

[0081] "Isolated antibody" refers to the purified state of the binding compound, meaning in such context that the molecule is substantially free of other biological molecules, such as nucleic acids, proteins, lipids, carbohydrates, or other substances, such as cell debris and growth medium. In general, the term "isolated" is not intended to refer to the complete absence of such substances, or the absence of water, buffers, or salts, so long as they are present in amounts that do not substantially interfere with the experimental or therapeutic uses of the binding compounds described herein.

[0082] The term "monoclonal antibody" as used herein refers to a population of substantially homogeneous antibodies, i.e., the antibody molecules comprising the population are identical in amino acid sequence, except for naturally occurring mutations that may be present in minor amounts. In contrast, conventional (polyclonal) antibody preparations typically contain a large number of different antibodies with different amino acid sequences in their variable domains, particularly in their CDRs, which are often specific for different epitopes. The modifier "monoclonal" indicates the character of the antibody as obtained from a substantially homogeneous population of antibodies, and this modifier should not be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies to be used according to the present invention can be produced by the hybridoma method first described by Kohler et al. (1975) Nature 256: 495, or can be produced by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). "Monoclonal" antibodies can also be isolated from phage antibody libraries using the techniques described, for example, in Clackson et al. (1991) Nature 352: 624-628 and Marks et al. (1991) J. Mol. Biol. 222: 581-597. See also Presta (2005) J. Allergy Clin. Immunol. 116:731.

[0083] As used herein, a "chimeric antibody" is an antibody having a variable domain from a first antibody and a constant domain from a second antibody, where the first and second antibodies are from different species. (U.S. Patent 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 from an experimental animal such as a rodent (the "parent antibody"), and the constant domain sequences are obtained from a human antibody; thus, the resulting chimeric antibody will be less likely to provoke an adverse immune response in a human subject than the parent (e.g., rodent) antibody.

[0084] As used herein, the term "humanized antibody" refers to a form of antibody that contains sequences from both human and non-human (e.g., mouse, rat) antibodies. In general, a humanized antibody will contain substantially all of at least one, typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the framework (FR) regions are those of a human immunoglobulin sequence. A humanized antibody may optionally contain at least a portion of a human immunoglobulin constant region (Fc).

[0085] The term "fully human antibody" refers to an antibody that contains only human immunoglobulin protein sequences. A fully human antibody may contain mouse glycochains if produced in a mouse, in a mouse cell, or in a hybridoma derived from a mouse cell. Similarly, a "mouse antibody" refers to an antibody that contains only mouse immunoglobulin sequences. Alternatively, a fully human antibody may contain rat glycochains if produced in a rat, in a rat cell, or in a hybridoma derived from a rat cell. Similarly, a "rat antibody" refers to an antibody that contains only rat immunoglobulin sequences.

[0086] Generally, the basic antibody structural unit comprises a tetramer. Each tetramer comprises two identical pairs of polypeptide chains, each pair having one "light" chain (about 25 kDa) and one "heavy" chain (about 50-70 kDa). The amino-terminal portion of each chain comprises a variable region of about 100-110 or more amino acids primarily responsible for antigen recognition. The carboxy-terminal portion of the heavy chain may define a constant region primarily responsible for effector function. Typically, human light chains are classified as kappa and lambda light chains. Furthermore, human heavy chains are typically classified as mu, delta, gamma, alpha or epsilon, defining the antibody's isotype as IgM, IgD, IgG, IgA and IgE, respectively. The variable and constant regions within the light and heavy chains are joined by a "J" region of about 12 or more amino acids, with the heavy chain also comprising a "D" region of about 10 amino acids. See generally, Fundamental Immunology Ch. 7 (Paul, W., ed., 2nd ed. Raven Press, NY (1989)).

[0087] The variable regions of each light / heavy chain pair form the antibody binding site. Thus, an intact antibody generally has two binding sites. Except in bifunctional or bispecific cases, the two binding sites are generally the same.

[0088] 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, which allow binding to a specific epitope. Generally, from N-terminus to C-terminus, both light and heavy chain variable domains contain FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. The assignment of amino acids to each domain is generally described in Sequences of Proteins of Immunological Interest, Kabat, et al.;National Institutes of Health, Bethesda, Md.;5th 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 Chothia, et al., (1989) Nature 342:878-883.

[0089] As used herein, the term "hypervariable region" refers to the amino acid residues of an antibody that are involved in antigen binding. The hypervariable region comprises 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; see Kabat et al. (1991) Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, US). Md.) and / or 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" residues 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 the antibody to an antigen or epitope. CDRs of interest are derived from donor antibody variable heavy and light chain sequences and include analogs of naturally occurring CDRs, which also share or retain the same antigen binding specificity and / or neutralizing ability as the donor antibody from which they were derived.

[0090] Additionally, in some embodiments, the antibody is a full length antibody, a single domain antibody, a recombinant heavy chain only antibody (VHH), a single chain antibody (scFv), a shark heavy chain only antibody (VNAR), a microprotein (cysteine ​​knot protein, knottin), DARPin; tetranectin; affibody; transbody; anticalin; AdNectin; affilin; microbody; peptide aptamer; alterase; plastic antibody; phylomer; stradobody; maxibody; shrimpbody; finomer, armadillo repeat protein, Kunitz domain, avimer, atrimer, probody, immunobody, triomab, troibody; pepbody; vaccibody, UniBody; affimer; DuoBody, Fv, Fab, Fab', F(ab')2, a peptidomimetic molecule, or a peptide mimetic molecule as described in U.S. Pat. No. 7,417,130, U.S. Patent Application Publication No. 2004 / 1133634. 32094, U.S. Patent No. 5,831,012, U.S. Patent Application Publication No. 2004 / 023334, U.S. Patent No. 7,250,297, U.S. Patent No. 6,818,418, U.S. Patent Application Publication No. 2004 / 209243, U.S. Patent No. 7,838,629, U.S. Patent No. 7,186,524, U.S. Patent No. 6,004,746, U.S. Patent No. 5,475,096, U.S. Patent Application Publication No. 2004 / 146938, U.S. Patent Application Publication No. The medicament may take the form of a synthetic molecule as described in U.S. Patent Application Publication No. 2004 / 157209, U.S. Patent No. 6,994,982, U.S. Patent No. 6,794,144, U.S. Patent Application Publication No. 2010 / 239633, U.S. Patent No. 7,803,907, U.S. Patent Application Publication No. 2010 / 119446, and / or U.S. Patent No. 7,166,697, each of which is hereby incorporated by reference in its entirety. See also Storz MAbs. 2011 May-Jun;3(3): 310-317, which is hereby incorporated by reference.

[0091] The term "antigen" as used herein means any molecule capable of raising or binding to an antibody, either directly or indirectly. The definition of "antigen" includes protein-encoding nucleic acid. "Antigen" may also refer to a binding partner of an antibody. In some embodiments, the antigen is an IGF-1R protein expressed on the surface of a cell. In some embodiments, the cell is an intact cell. An intact cell is a cell that has not been lysed or ruptured by the use of 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, a method for producing an antibody that binds to an IGF-1R protein is provided herein, the method comprising culturing a cell that contains a nucleic acid molecule encoding an IGF-1R antibody.

[0092] As used herein, "specific binding" or "immunospecific binding" or "immunospecifically binds" refers to an antibody that binds to a given antigen (e.g., IGF-1R) or binds to an epitope present on an antigen. In some embodiments, the antibody is -7 M or less dissociation constant (K D ) and its K for binding to a non-specific antigen other than the predetermined antigen (e.g., BSA, casein, or another non-specific polypeptide) D K is less than half of D The phrases "antibody that recognizes IGF-1R" and "antibody specific for IGF-1R" are used herein synonymously with the term "antibody that immunospecifically binds to IGF-1R". The present disclosure may refer 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 purpose. In some embodiments, the antibody, or binding compound derived from the antigen-binding site of the antibody, of the contemplated method binds to its antigen (IGF-1R) with an affinity at least 2-fold greater, at least 10-fold greater, at least 20-fold greater, or at least 100-fold greater than the affinity for any other antigen.

[0093] Methods for determining mAb specificity and affinity by competitive inhibition can be found 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.

[0094] The term "homolog" refers to a protein sequence having between 40% and 100% sequence homology or identity to a reference sequence. The percent identity between two peptide chains can be determined by pairwise alignment using default settings in the AlignX module of Vector NTI v.9.0.0 (Invitrogen Corp., Carslbad, Calif). 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% homology or identity to a sequence described herein. In some embodiments, the antibody has conservative substitutions compared to the sequences described herein. Exemplary conservative substitutions are shown in Table 1 and are encompassed within the scope of the disclosed subject matter. Conservative substitutions may be present in framework regions or in the antigen-binding site as long as they do not adversely affect the properties of the antibody. Substitutions can be made to improve the properties of the antibody, such as stability or affinity. Conservative substitutions will result in a molecule that has similar functional and chemical properties as the molecule without such alteration. Exemplary amino acid substitutions are shown in the table below. [Table 1]

[0095] In some embodiments, variants of the proteins and peptides provided herein are provided. In some embodiments, the variants include substitutions, deletions, or insertions. In some embodiments, the variants include 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 (e.g., 1-10) substitutions. As described herein, the substitutions may be conservative substitutions. In some embodiments, the substitutions are non-conservative substitutions. In some embodiments, the variants include 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 (e.g., 1-10) deletions. In some embodiments, the variants include 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 defined herein. In some embodiments, the substitutions, deletions, or insertions are not present in the CDRs defined herein.

[0096] The term "in combination with" as used herein means that the agents described can be administered to an animal or subject together in a mixture, simultaneously as a single agent, or sequentially in any order as a single agent.

[0097] Techniques for raising antibodies against small peptide sequences that recognize and bind to those sequences when presented as native sequences in free or conjugated form or in the context of large 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 human, mouse, sheep, rat, rabbit, shark, llama or chicken. In some embodiments, antibodies are produced in chicken. Antibodies can also be produced in other small animals.

[0098] The term "epitope" is intended to refer to the portion of any molecule that an antibody can recognize or bind to at one or more of the antigen-binding regions of the Ab. Epitopes usually consist of chemically active surface groups 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 non-denatured protein. In some embodiments, the epitope is present only in denatured protein.

[0099] 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.

[0100] The hybrid cells are formed by fusion of non-human antibody-producing cells, typically spleen cells of an animal that is immune to either the natural or recombinant antigen, or a peptide fragment 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 an animal immunized with the antigen.

[0101] The second fusion partner that provides the immortalization function can be lymphoblastoid cells or plasmacytoma or myeloma cells, which are not themselves antibody-producing cells and are malignant. Fusion partner cells include, but are not limited to, hybridoma SP2 / 0-Ag14 (ATCC CRL1581), abbreviated as SP2 / 0, and myeloma P3X63Ag8 (ATCC TIB9), or derivatives thereof. See, for example, Ausubel, infra; Harlow, infra; and Colligan, infra, the contents of which are incorporated herein by reference in their entirety.

[0102] Antibodies can be generated according to the examples provided herein. Once the sequence is known, the antibodies can also be generated according to known methods. Antibodies can also be converted to a different type, such as to a human IgG. By converting the antibody to a human antibody, the human subject should not see the antibody as foreign. Conversion of non-human IgG antibodies to human IgG antibodies is well known and can be done routinely once the native sequence is known. Antibodies can be modified according to known methods, as discussed herein. 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 that contribute nucleotide sequences encoding the antigen-binding region of a chimeric antibody can also be produced by transformation of non-human, e.g., primate, or human cells. For example, antibody-producing B lymphocytes can be infected and transformed with a virus, such as Epstein-Barr virus, to give rise to 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 in the art. See, e.g., Ausubel, infra, Harlow, infra, and Colligan, infra, the contents of which are incorporated herein by reference in their entireties. Cell fusion is accomplished 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, infra, Harlow, infra, and Colligan, infra, the contents of which are incorporated herein by reference in their entireties.

[0103] In some embodiments, the antibody is a MAb that binds to IGF-1R, hi some embodiments, the antibody binds to an amino acid of an epitope of IGF-1R.

[0104] In some embodiments, the antibody comprises a sequence defined herein.

[0105] The antibody sequence can be modified to produce a human IgG antibody. The conversion of the sequences provided herein can be modified to produce other types of antibodies. The CDRs can also be linked to other antibodies, proteins or molecules to generate antibody fragments that bind to IGF-1R. This can be in the form of an antibody drug conjugate ("ADC"), a multispecific molecule, or a chimeric antigen receptor. The CDRs and antibody sequences provided herein can also be humanized or made fully human according to known methods. The sequences can also be made into chimeric antibodies as described herein.

[0106] In some embodiments, the antibody comprises an amino acid sequence comprising a sequence defined herein or a fragment thereof. In some embodiments, the antibody comprises one or more amino acid sequences as provided herein, an antigen-binding fragment thereof, or a human IgG variant thereof. "Human IgG variant thereof" refers to an antibody that has been modified to become a human IgG when the starting antibody is not a human IgG antibody.

[0107] As described herein, the production of antibodies with known sequences is routine and can be performed by any method. Thus, in some embodiments, a nucleic acid is provided that encodes an antibody or a fragment thereof. In some embodiments, the nucleic acid encodes a sequence as defined herein. The antibody can also be modified to be a chimeric or human antibody. The antibody can also be used in an injectable pharmaceutical composition. Also, as described herein, the antibody can be an isolated antibody or an engineered antibody.

[0108] In some embodiments, "derivatives" of antibodies, fragments, regions or derivatives thereof are provided, which term includes proteins encoded by genes that have been truncated or modified to produce molecular species that resemble the immunoglobulin fragments in function. Modifications include, but are not limited to, the addition of gene sequences that code for toxic proteins, such as plant and bacterial toxins. Modifications may also include reporter proteins, such as fluorescent or chemiluminescent tags. Fragments and derivatives can be produced by any method.

[0109] 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 utilities comparable to the embodiments of the present application.

[0110] The nucleic acid sequence encoding the antibody described herein may be genomic or cDNA or RNA (e.g., mRNA) encoding at least one of the variable regions described herein. A convenient alternative to the use of chromosomal gene fragments as a source of DNA encoding V-region antigen-binding segments is the use of cDNA for the construction of 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), which references are hereby incorporated by reference in their entirety). The use of cDNA requires gene expression elements suitable for the host cell to be combined with the gene to achieve synthesis of the desired protein. The use of cDNA sequences has an advantage over genomic sequences (containing introns) in that the cDNA sequences can be expressed in bacteria or other hosts that lack a suitable RNA splicing system.

[0111] For example, cDNAs encoding V-region antigen-binding segments capable of detecting, binding to, or neutralizing IGF-1R antigens can be obtained using known methods based on the use of the amino acid sequences provided herein. Because the genetic code is degenerate, more than one codon can be used to code for a particular amino acid (Watson, et al., infra). Using the genetic code, one or more different oligonucleotides can be identified, each of which could code for an amino acid. The probability that a particular oligonucleotide actually constitutes an actual XXX coding sequence can be estimated by considering unusual base pairing relationships and the frequency with which a particular codon is actually used (to code for a particular amino acid) in a eukaryotic or prokaryotic cell expressing the antibody or fragment. Such "codon usage rules" are disclosed by Lathe, et al., J. Molec. Biol. 183:1 12 (1985). Using Lathe's "codon usage rules," a single oligonucleotide or set of oligonucleotides is identified that contains the theoretically "most likely" nucleotide sequence that can code for an antibody variable or constant region sequence.

[0112] 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 derived from human fetal liver libraries by any method. Human C region genes can be derived from any human cell, including those that express and produce human immunoglobulins. Human C H The regions may be derived from any of the known classes or isotypes of human H chains, including gamma, μ, α, δ, or ε, as well as subtypes thereof, such as G1, G2, G3, and G4. The H chain isotype is responsible for the various effector functions of the antibody, and therefore the C H The choice of region will be guided by the desired effector function, e.g., complement fixation, or antibody-dependent cellular cytotoxicity (ADCC) activity. HThe regions are derived from gamma 1 (IgG1), gamma 3 (IgG3), gamma 4 (IgG4), or μ (IgM). L The region may be derived from either human light chain isotype, kappa or lambda. In some embodiments, the antibody comprises an Fc 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 hereby incorporated by reference in its entirety. In some embodiments, the constant region comprises a mutation at amino acid residue 428, as compared to a wild-type human IgG constant domain, numbered according to the EU numbering index of Kabat. Without being bound to any particular theory, an antibody 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 a substitution of the native residue with threonine, leucine, phenylalanine, or serine. In some embodiments, the antibody further comprises one or more amino acid substitutions at one or more of amino acid residues 251-256, 285-290, 308-314, 385-389, and 429-436 according to the Kabat EU numbering index, relative to the corresponding wild-type human IgG constant domain. Specific mutations or substitutions at these positions are described in U.S. Patent No. 7,670,600, which is hereby incorporated by reference in its entirety.

[0113] Genes encoding human immunoglobulin C regions can be obtained from human cells by 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 are readily available from known clones that contain genes representing the two classes of L chains, the five classes of H chains, and their subclasses. Chimeric antibody fragments, such as F(ab')2 and Fab, can be prepared by designing chimeric H chain genes that are appropriately truncated. For example, a chimeric gene encoding the H chain portion of the F(ab')2 fragment will contain DNA sequences encoding the CH1 domain and hinge region of the H chain, followed by a translation stop codon to generate a truncated molecule.

[0114] In some embodiments, the antibodies, murine, human, humanized or chimeric antibodies, fragments and regions of the antibodies 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 synthesizing these DNA segments in a C H and C L These are produced by combining DNA segments encoding these regions, respectively, to generate mouse, human, or chimeric immunoglobulin-encoding genes.

[0115] Thus, in some embodiments, a fusion chimeric gene is created that includes 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, linked to a second DNA segment encoding at least a portion of a human C region.

[0116] Thus, the method of producing antibodies according to some of the embodiments described herein, including cDNA encoding antibody V and C regions, comprises several steps as exemplified below: 1. Isolation of messenger RNA (mRNA) from a cell line producing an anti-IGF-1R antigen antibody, and from an additional antibody as needed to provide heavy and light chain constant regions; cloning and producing cDNA therefrom; 2. Preparation of a full-length cDNA library from the purified mRNA, which can be (i) identified with a suitable probe, (ii) sequenced, and (iii) matched with a C or V gene segment from another antibody for chimeric antibodies; 3. Construction of a complete H or L chain coding sequence by ligation of a particular cloned V region gene segment to the cloned C region gene described above; 4. Expression and production of L and H chains in a selected host, including prokaryotic and eukaryotic cells, to provide mouse-mouse, human-mouse, human-human or human-mouse antibodies.

[0117] Two coding DNA sequences are said to be "operably linked" if the linkage results in a continuously translatable sequence without altering or disrupting the triplet reading frame. A DNA coding sequence is operably linked to a gene expression element if the linkage results in the proper function of the gene expression element leading to expression of the coding sequence.

[0118] As used herein, and unless otherwise indicated, the term "about" is intended to mean ±5% of the value it modifies. Thus, about 100 means from 95 to 105.

[0119] 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 for detecting the presence of an antigen.

[0120] The term "purified" with respect to an antibody refers to an antibody that is substantially free of other materials that accompany the molecule in its natural environment. For example, a purified protein is substantially free of cellular material and other proteins from the cell or tissue from which it is derived. The term refers to preparations in which the isolated protein is sufficiently pure to be analyzed, 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, the antibody is purified.

[0121] As an alternative to preparing hybridomas that secrete monoclonal antibodies, 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 polypeptide described herein, thereby isolating 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. In addition, examples of methods and reagents that are 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 for therapy, diagnosis, or as research tools.

[0122] Antibody conjugates

[0123] The antibody as defined herein can also be conjugated to a chemical moiety. The chemical moiety can be, inter alia, 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 half-life of the antibody molecule in the subject's body. Suitable polymers include, but are not limited to, polyethylene glycol (PEG) (e.g., PEG with a molecular weight of 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 drugs, such as calicheamicins (e.g., ozogamicin), monomethyl auristatin E, mertansine, and the like. Other examples include, but are not limited to, bioactive microtubule inhibitors, alkylating agents, and DNA minor groove binders. Other examples are provided herein below. Chemical moieties can be linked to antibodies by linking groups (maleimides), cleavable linkers, such as cathepsin-cleavable linkers (valine-citrulline), and in some embodiments, one or more spacers (e.g., para-aminobenzyl carbamates). Without being bound to any particular theory, when the antibody conjugate binds to IGF-1R it can be internalized and the chemical moiety can kill or otherwise inhibit the growth of the cell. In some embodiments, the cell is a thyroid cell.

[0124] The antibodies and antibody fragments of the invention can be labeled with a label, e.g. 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 can also be conjugated with Fe.

[0125] Antibodies and antibody fragments can be coupled to fluorophores, such as rare earth chelates, fluorescein and its derivatives, rhodamine and its derivatives, isothiocyanates, phycoerythrin, phycocyanin, allophycocyanin, o-phthaladehyde, fluorescamine, 152 They may also be conjugated with fluorescent or chemiluminescent labels, including 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.

[0126] The antibody molecule may also be conjugated to a cytotoxic agent, 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 protein, Phytoiacca americana proteins PAPI, PAPII, and PAP-S, momordica charantia inhibitor, curcin, crotin, saponaria officinalis inhibitor, mitogenin, restrictocin, phenomycin, and enomycin.

[0127] To conjugate the antibody molecules of the present invention to various moieties, any method known in the art can be utilized, including those described in 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.

[0128] Chimeric Antigen Receptor

[0129] The antibodies provided herein can also be incorporated into chimeric antigen receptors ("CARs"), which 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 so that they will attack cells that express IGF-1R. T cells are taken from the patient's blood. A special receptor gene that binds to a certain 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 defined herein. CAR-T cells containing the IGF-1R antibody can then be used to treat conditions such as those defined herein.

[0130] In some embodiments, an antibody (e.g., an anti-IGF-1R antibody) is provided herein. In some embodiments, the antibody is a recombinant antibody that binds to IGF-1R protein. In some embodiments, the IGF-1R protein is a human IGF-1R protein. In some embodiments, the IGF-1R protein recognized by the antibody is in its native conformation (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 is not naturally occurring. In some embodiments, the term "recombinant antibody" refers to an antibody that is not isolated from a human subject.

[0131] In some embodiments, the antibody comprises one or more peptides having the following sequences, or variants thereof: [Table 3]

[0132] The VH and VL sequences may be of any format, including but not limited to an scFv format in which the VH and VL regions are linked by a peptide linker. An example of a peptide linker that may be used to link the various peptides defined herein is (GGGGS) n (SEQ ID NO: 12); (GGGGA) n (SEQ ID NO: 13), or any combination thereof, where each n is independently 1 to 5. In some embodiments, the variable regions are not linked by a peptide linker. In some embodiments, the antibody comprises or consists of a polypeptide set forth in SEQ ID NOs: 10 and 11. In some embodiments, the antibody comprises a polypeptide including SEQ ID NOs: 3, 4, 5, 6, 7, 8, and 9.

[0133] 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.

[0134] [Table 4]

[0135] In some embodiments, the antibody comprises one or more peptides having the following sequence, or a variant thereof, including one or more variable domains (italics), CDRs (italics and bold), and a human IgG1 / kappa constant domain (underlined): [Table 5]

[0136] In some embodiments, the antibody, or antibody binding fragment thereof, comprises a heavy or light chain having the sequence of SEQ ID NO: 10 and 11. In some embodiments, the antibody, or antibody binding fragment thereof, comprises a heavy chain having the sequence of SEQ ID NO: 10. In some embodiments, the antibody, or antibody binding fragment thereof, comprises a heavy chain having a sequence that is 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% or substantially 100% identical to that of SEQ ID NO: 10. In some embodiments, the sequence that is 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% or substantially 100% identical to that of SEQ ID NO: 10 comprises the CDRs of SEQ ID NOs: 7, 8 and / or 9 shown above.

[0137] In some embodiments, the antibody, or antibody binding fragment thereof, comprises a light chain having the sequence of SEQ ID NO: 11. In some embodiments, the antibody, or antibody binding fragment thereof, comprises a light chain having a sequence that is 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% or substantially 100% identical to that of SEQ ID NO: 11. In some embodiments, the sequence that is 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% or substantially 100% identical to that of SEQ ID NO: 11 comprises the CDRs of SEQ ID NOs: 7, 8 and / or 9 shown above.

[0138] In some embodiments, the antibody, or antibody binding fragment thereof, comprises a light chain CDR having the sequence of SEQ ID NO: 4, 5 or 6. In some embodiments, the antibody, or antibody binding fragment thereof, comprises a heavy chain CDR having the sequence of SEQ ID NO: 7, 8 or 9.

[0139] In some embodiments, the antibody, or antibody-binding fragment thereof, comprises a light chain having LCDR1, LCDR2 and LCDR3, wherein LCDR1 has the sequence of SEQ ID NO:4, LCDR2 has the sequence of SEQ ID NO:5, and LCDR3 has the sequence of SEQ ID NO:6.

[0140] 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:7, HCDR2 has the sequence of SEQ ID NO:8, and HCDR3 has the sequence of SEQ ID NO:9.

[0141] Different CDR motifs can be combined in any combination, including those not depicted in the table above. For example, the following embodiments are provided as non-limiting examples of such combinations.

[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, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 6; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2 and CDR3, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 7, the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 8, and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 9; or any variant 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, wherein the light chain CDR1 sequence has an amino acid sequence that is 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% or substantially 100% identical to that of SEQ ID NO:4, the light chain CDR2 has an amino acid sequence that is 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% or substantially 100% identical to that of SEQ ID NO:5, and the light chain CDR3 sequence has an amino acid sequence that is 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% or substantially 100% identical to that of SEQ ID NO:6; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2 and CDR3, wherein the heavy chain CDR1 sequence has an amino acid sequence that is 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% or substantially 100% identical to that of SEQ ID NO:7, the heavy chain CDR2 sequence has an amino acid sequence that is 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% or substantially 100% identical to that of SEQ ID NO:8, and the heavy chain CDR3 sequence has an amino acid sequence that is 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% or substantially 100% identical to that of SEQ ID NO:9; or any variant of the foregoing.

[0144] In some embodiments, an antibody, or antigen-binding fragment thereof, or protein is provided that comprises a peptide having a sequence set forth in any of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11.

[0145] In some embodiments, the antibody, or antigen-binding fragment thereof, comprises a sequence of any of those mentioned above, or a variant of any of those mentioned above.

[0146] Pharmaceutical Compositions

[0147] 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 pharma- ceutically acceptable carrier or excipient.See, e.g., Remington's Pharmaceutical Sciences and US Pharmacopeia: National Formulary, Mack Publishing Company, Easton, PA (1984).

[0148] Formulations of the therapeutic and diagnostic agents can be prepared by mixing with acceptable carriers, excipients or stabilizers, for example, in the form of a lyophilized powder, a slurry, an aqueous solution or an aqueous 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, and NaCl or sucrose is added for isotonicity. Additional agents, such as polysorbate 20 or polysorbate 80, may be added to enhance stability.

[0149] The toxicity and therapeutic efficacy of the antibody composition, administered alone or in combination with another agent, can be determined, for example, by the LD 50 (a 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 cultures or experimental animals. The dose ratio between toxic and therapeutic effects is the therapeutic index (LD 50 / ED 50 In certain embodiments, antibodies that exhibit high therapeutic indices are desirable. The data obtained from these cell culture assays and animal studies can be used in formulating a range of dosages for use in humans. The dosage of such compounds is preferably within the ED 50 The dosage may vary within this range depending upon the dosage form utilized, and the route of administration.

[0150] In some embodiments, compositions of the invention are administered to a subject in accordance with the Physicians' Desk Reference 2003 (Thomson Healthcare; 57th edition (November 1, 2002)).

[0151] The method of administration can be varied. Suitable routes of administration include oral, rectal, transmucosal, intestinal, parenteral, intramuscular, subcutaneous, intradermal, intramedullary, intrathecal, direct intracerebroventricular, intravenous, intraperitoneal, intranasal, intraocular, inhalation, insufflation, topical, dermal, transdermal or intra-arterial routes.

[0152] In some embodiments, the antibody or antigen-binding fragment thereof can be administered by invasive route, for example, by 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, aerosol delivery. Administration by non-invasive route (e.g., orally; e.g., by pill, capsule, or tablet) is also within the scope of this embodiment.

[0153] 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, any therapeutic agent used to treat thyroid eye disease or a condition related thereto. Examples of such treatments and therapeutic agents include antithyroid drugs, diabetes drugs, 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, acebuside, ce ... In some embodiments, the medicament may be administered intravenously orally, for example, by administering to the patient an effective amount of the medicament. In some embodiments, the medicament may be administered intravenously orally, for example, by administering to the patient an effective amount of the medicament. In some embodiments, the medicament may be administered intravenously orally, for example, by administering to the patient an effective amount of the medicament.

[0154] The compositions can be administered with medical devices known in the art, for example, pharmaceutical compositions of the invention can be administered by injection with a hypodermic needle, including, for example, a prefilled syringe or an autoinjector.

[0155] The pharmaceutical compositions can also be administered with a needleless hypodermic injection device, such as those disclosed in U.S. Pat. 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 4,596,556.

[0156] Pharmaceutical compositions can also be administered by injection. Examples of well-known implantable devices and module forms for administering pharmaceutical compositions include U.S. Patent No. 4,487,603, which discloses an implantable microinfusion pump for dispensing therapeutic agents at a controlled rate; U.S. Patent No. 4,447,233, which discloses a therapeutic agent infusion pump for delivering drugs at a precise infusion rate; U.S. Patent No. 4,447,224, which discloses a variable flow rate implantable infusion device for continuous drug delivery; U.S. Patent No. 4,439,196, which discloses an osmotic drug delivery system with multiple compartments. Many other such implantable devices, delivery systems, and modules are known to those skilled in the art.

[0157] Alternatively, the antibody can be administered locally, rather than systemically, for example, by injection of the antibody, often in a depot or sustained release formulation, directly into arthritic joints or pathogen-induced lesions characterized by immunological pathology.Furthermore, the antibody can be administered in a targeted drug delivery system, for example, in liposomes coated with tissue-specific antibodies, which target, for example, arthritic joints or pathogen-induced lesions characterized by immunological pathology.The liposomes will be targeted to and selectively taken up by the affected tissue.

[0158] The administration regimen depends on several factors, including the serum or tissue turnover rate of the therapeutic antibody, the level of symptoms, the immunogenicity of the therapeutic antibody, and the accessibility of the target cells in the biological matrix.Preferably, the administration regimen delivers sufficient therapeutic antibody to simultaneously cause improvement of the target disease state while minimizing undesirable side effects.Thus, the amount of biologic delivered depends in part on the specific therapeutic antibody and the severity of the condition being treated. Guidance regarding the selection of 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 (see, e.g., Ghosh et al. (2003) New Engl. J. Med. 348:24-32; Lipsky et al. (2000) New Engl. J. Med. 343:1594-1602).

[0159] The determination of the appropriate dose is made by the clinician, for example, using parameters or factors known or believed in the art to affect treatment.Generally, the dose is started at a somewhat lower amount than the optimal dose, and then increased in small increments until the desired or optimal effect is achieved relative to any negative side effects.Important diagnostic measures include symptoms, for example, symptoms of inflammation, or the level of inflammatory cytokines produced.Generally, it is desirable that the biologics to be used are derived from the same species as the animal that is the target of treatment, thereby minimizing any immune response to the reagent.For human subjects, for example, chimeric, humanized and fully human antibodies are desirable.

[0160] The antibody or antigen-binding fragment thereof can be provided by continuous infusion or by a dose administered, for example, once daily, 1-7 times per week, once per week, once every two weeks, once per month, once every two months, quarterly, twice per year, annually, etc. The dose can be provided, for example, intravenously, subcutaneously, topically, orally, nasally, rectally, intramuscularly, intracerebrally, intraspinally, or by inhalation. The total weekly dose will generally be 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 (see, 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 (See Immunol. Immunother. 52:133-144). 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 or higher. In other embodiments, fully human antibodies are administered subcutaneously or intravenously at 10, 20, 50, 80, 100, 200, 500, 1000 or 2500 mg / subject once a week, once every two weeks, "every four weeks", once a month, once every two months or quarterly, or as otherwise defined herein.

[0161] As used herein, "suppress" or "treat" or "treatment" includes delaying the onset of symptoms associated with a disorder and / or reducing the severity of symptoms of such disorders. These terms further include alleviating existing uncontrolled or undesirable symptoms, preventing additional symptoms, and alleviating or preventing the underlying disease of such symptoms. Thus, these terms mean that beneficial results are achieved in a vertebrate subject having a disorder, disease or condition or a vertebrate subject that may develop such a disorder, disease or condition.

[0162] 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 is effective when administered alone or in combination with an additional therapeutic agent to a cell, tissue or subject 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 cause at least partial alleviation of symptoms, such as treating, curing, preventing or alleviating an associated medical condition, or increasing the treatment, cure, prevention or alleviation rate 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 active ingredients that produces a therapeutic effect, whether administered sequentially or simultaneously in combination. An effective amount of a therapeutic agent results in an improvement in a diagnostic measure or parameter by 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 a subjective scale when the subjective scale is 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 ameliorate a condition as defined herein.

[0163] The term "subject" as used throughout includes any organism, such as an animal, including mammals (e.g., rats, mice, dogs, cats, rabbits) and, for example, humans. A subject may also be referred to as a patient. In some embodiments, a subject is a subject in need of treatment. A subject "in need of treatment" refers to a subject who has been identified as needing treatment for a condition to be treated, and who is treated with the specific intent of treating such condition. The condition may be, for example, any of the conditions described herein.

[0164] The isolated antibodies bind to an epitope on the IGF-1R protein or other proteins described herein and exhibit in vitro and / or in vivo IGF-1R inhibitory or therapeutic activity, while the antibodies or antigen-binding fragments thereof capable of inhibiting IGF-1R function are suitable as therapeutic agents for treating IGF-1R-related conditions in humans and in animals. These conditions include thyroid eye disease. Thus, methods of treating such conditions are also provided, comprising administering the antibody or antigen-binding fragment thereof to a subject having such a condition.

[0165] In some embodiments, the methods include administering a therapeutically or prophylactically effective amount of one or more monoclonal antibodies or antigen-binding fragments of the antibodies described herein to a susceptible subject or to a person exhibiting a condition known or suspected to be IGF-1R caused pathology observed. 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 antibodies defined herein.

[0166] As used herein, IGF-1R-related pathology refers to conditions caused by modulation of IGF-1R, including, but not limited to, thyroid eye disease and other conditions defined herein.

[0167] In some embodiments, the antibodies used are compatible with the recipient species, such 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.

[0168] Treatment of an individual may include administering a therapeutically effective amount of the 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 a mixture with another therapeutic, analgesic or diagnostic agent, such as those defined herein.When providing a patient with an antibody or its fragment that can bind to IGF-1R, or an antibody that can provide protection from IGF-1R pathology in the recipient patient, the dosage of the agent administered will vary depending on factors such as the patient's age, weight, height, sex, general medical condition, previous medical history, etc.

[0169] Antibodies capable of treating conditions associated with IGF-1R activity, or for use in treating IGF-1R-related pathologies, are intended to be provided to a subject in an amount sufficient to affect the reduction, elimination or amelioration of an IGF-1R-related symptom or pathology, including thyroid eye disease, and the like.

[0170] Thus, in some embodiments, a method for treating a subject having an IGF-1R-related disorder is provided.In some embodiments, the method comprises administering a pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof as provided herein.In some embodiments, the disorder is thyroid eye disease.As defined herein, the antibody or an antigen-binding fragment thereof can be administered together with other therapeutic agents.They can be administered simultaneously or sequentially.

[0171] In some embodiments, the antibodies or antigen-binding fragments thereof can be used to treat thyroid eye disease. In some embodiments, the antibodies or antigen-binding fragments thereof can be used to treat or reduce the severity of thyroid eye disease (TAO) or its symptoms.

[0172] In some embodiments, a method or use is provided for reducing exophthalmos in an eye of a subject with thyroid eye disease (TAO).

[0173] In some embodiments, the subject is one who has been previously treated with an antibody different from those provided herein.

[0174] In some embodiments, the method or use provides for a Clinical Activity Score (CAS) in a subject having or suspected of having Thyroid Eye Disease (TAO).

[0175] In some embodiments, the method or use is provided to reduce exophthalmos by at least 2 mm. In some embodiments, the method or use is provided to reduce exophthalmos by at least 3 mm. In some embodiments, the method or use is provided to reduce exophthalmos by at least 2-3 mm or 2-4 mm. In some embodiments, exophthalmos is reduced by at least 2, 3 or 4 mm. In some embodiments, the reduction in exophthalmos is seen within 3 weeks of administration of the first dose. In some embodiments, the reduction in exophthalmos is seen within 6 weeks of administration of the first dose.

[0176] In some embodiments, the subject has a reduced Clinical Activity Score (CAS) in Subjects with Thyroid Eye Disease (TAO).

[0177] As used herein, the term Clinical Activity Score (CAS) refers to a protocol described and scored according to Table 2. According to this protocol, one point is given for the presence of each parameter evaluated in the table below. The sum of all points defines clinical activity and gives the CAS, where 0 or 1 corresponds to inactive disease and 7 corresponds to severe active ophthalmopathy. [Table 2]

[0178] As provided in Table 2, the CAS consists of seven components: spontaneous retrobulbar pain, pain on attempted eye movement (upward, left, right, and downward gaze), conjunctival redness, eyelid redness, chemosis, swelling of the caruncle / fold, and swelling of the eyelid. Each component is scored as present (score 1) or absent (score 0). The score at each efficacy assessment is the sum of all items present, ranging from 0 to 7, where 0 or 1 corresponds to inactive disease and 7 corresponds to severe active ophthalmopathy. A change of >2 points is considered clinically meaningful. In some embodiments, the subject's score improves by at least 2, 3, or 4 points. In some embodiments, the subject's score improves within 3 weeks of the first dose. In some embodiments, the subject's score improves within 6 weeks of the first dose.

[0179] Item 1, spontaneous orbital pain, can be a pain or pressure on or in the eye. This pain can be caused by increased intraorbital pressure when the orbital tissue volume increases due to excess synthesis of extracellular matrix, fluid accumulation, and cellular infiltration and expansion. Item 2, gaze-induced orbital pain, can be pain in the eye when looking up, down, or sideways, or when attempting to do so, i.e., pain associated with upward, downward, or lateral eye movement, or pain when attempting to move the eye. This type of pain can be caused by stretching of the inflamed muscles, especially when attempting to look upward. "Stretching pain" is not induced by digital pressure on the eye, as would be expected if it were a manifestation of elevated intraorbital pressure. Both types of pain can be relieved after anti-inflammatory treatment. These types of pain are therefore considered to be directly related to autoimmune inflammation in the orbit and therefore useful for assessing TAO activity.

[0180] Swelling in TAO is seen as chemosis (edema of the conjunctiva), item number 6 in Table 1, and swelling of the caruncle and / or semilunar fold. Both are signs of TAO activity. Eyelid swelling may be due to edema, fat herniation from the orbital septum, or fibrous degeneration. In addition to swelling, other symptoms suggestive of active TAO include redness and / or pain of the conjunctiva, eyelid, caruncle, and / or semilunar fold.

[0181] In some embodiments, the subject being treated has exophthalmos and it is reduced by at least 2 mm. In some embodiments, the subject being treated has exophthalmos and it is reduced by at least 3 mm. In some embodiments, the subject being treated has exophthalmos and it is reduced by at least 4 mm.

[0182] In some embodiments, the subject's Clinical Activity Score (CAS) is reduced by at least 2 points in the treated subject. In some embodiments, the subject's Clinical Activity Score (CAS) is reduced to one (1). In some embodiments, the subject's Clinical Activity Score (CAS) is reduced to zero (0).

[0183] In some embodiments, a method is provided for treating or reducing the severity of thyroid eye disease (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 2 mm or more in the other (or fellow) eye; and (iii) reduces CAS in the subject to one (1) or zero (0).

[0184] In some embodiments, methods are provided for improving quality of life in a subject with thyroid eye disease (TAO, also known as Graves' ophthalmopathy / Graves' orbitopathy). In some embodiments, quality of life is measured by a Graves' Ophthalmopathy Quality of Life (GO-QoL) assessment or by either its visual function or appearance subscales. In some embodiments, treatment results in an improvement of greater than or equal to 8 points on the GO-QoL. In some embodiments, treatment results in an improvement in the function subscale of the GO-QoL. In some embodiments, treatment results in an improvement in the appearance subscale of the GO-QoL.

[0185] In some embodiments, a method is provided for treating or reducing the severity of diplopia in a subject with thyroid eye disease (TAO). In some embodiments, the diplopia is constant diplopia. In some embodiments, the diplopia is non-constant diplopia. In some embodiments, the diplopia is intermittent diplopia. In some embodiments, the improvement in diplopia or the reduction in its severity is sustained at least 20 weeks after cessation of antibody administration. In some embodiments, the improvement in diplopia or the reduction in its severity is sustained at least 50 weeks after cessation of antibody administration. In some embodiments, the diplopia is improved in the subject within 3 weeks or within 6 weeks of the first dose.

[0186] Severity of the disease may be measured in the following non-limiting embodiments. For example, for eyelid opening, the distance between the eyelid margins is measured (in mm) with the patient sitting in a relaxed position and fixated at a distance, looking straight ahead. For eyelid swelling, the scale / 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 fold is either absent or present. Exophthalmos is measured in millimeters using the same Hertel exophthalmometer and the same intercanthal distance for each individual patient. Subjective diplopia is scored from 0 to 3 (0=no diplopia; 1=intermittent, i.e., diplopia in straight gaze position when fatigued or first awakened; 2=non-constant, i.e., diplopia in extreme gaze; 3=constant, i.e., continuous diplopia in straight gaze or reading position). For ocular muscle lesions, the twitching is measured in degrees. Corneal lesions are either absent / punctate or keratopathy / ulcer. For optic nerve lesions, i.e., best corrected visual acuity, color vision, optic disc, relative afferent pupillary defect, the status is either absent or present. In addition, visual fields are checked if optic nerve compression is suspected. In some embodiments, patients may be classified according to the following severity classifications: For example, vision-threatening thyroid eye disease: patients with thyroid optic neuropathy (DON) and / or corneal destruction. This category warrants immediate intervention. Moderate to severe thyroid eye disease: Patients without vision-threatening disease whose eye disease has sufficient impact on daily life to justify the risk of immunosuppression (if active) or surgical intervention (if inactive). Patients with moderate to severe thyroid eye disease usually have one or more of the following: eyelid retraction greater than or equal to 2 mm; moderate or severe soft tissue involvement; exophthalmos greater than or equal to 3 mm above normal for race and sex; non-constant or constant diplopia.Mild thyroid eye disease: Patients whose features of thyroid eye disease have minimal impact on daily life that is insufficient to justify immunosuppression or surgical procedures. They usually have only one or more of the following: slight eyelid retraction (<2 mm), mild soft tissue involvement, exophthalmos <3 mm above normal for race and sex, transient or absent diplopia, and corneal exposure that responds to lubricants.

[0187] In some embodiments, patients can be characterized by Graves' Ophthalmopathy Quality of Life (GO-QoL) score. In addition to exophthalmos (or exophthalmos) and CAS, quality of life is also assessed by using the GO Quality of Life (GO-QoL) questionnaire. This questionnaire is designed to determine the improvement of quality of life after treatment with the methods disclosed herein. In some embodiments, the questionnaire can determine 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. GO-QoL is a 16-item self-administered questionnaire divided into two subsets, and is used to evaluate the impact of TED as perceived by patients on (i) their daily physical activity as related to visual function, and (ii) psychosocial function, and quality of life is assessed by using the GO QoL questionnaire. The GO-QoL questionnaire [CB Terwee 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 assessment subscales, one covering the impact of visual function on daily activities, the other assessing the impact of self-perceived appearance. The visual function subscale covers activities such as driving, walking, reading, watching television, etc. The appearance subscale covers questions such as whether the ophthalmopathy has altered the subject's appearance, whether it causes other people to react negatively to the subject, whether it causes social isolation, and whether it causes the subject to try to hide their appearance. Each subscale has eight questions answered with high--very much; high--somewhat; or no--not at all. Each question is scored from 0 to 2, and then the total raw score is mathematically converted to a 0 to 100 scale, with 0 representing the worst impact on quality of life and 100 representing no impact. A change of >8 points, or greater than or equal to 8 points, on the 0 to 100 scale has been shown to be clinically meaningful. The composite score takes the raw scores from both subscales and converts them back to a single 0 to 100 scale.The questionnaire has two self-rating subscales. Each subscale has eight questions that are answered with (i) high--very much; (ii) high--somewhat; or (iii) no--not at all. Each question is scored from 0 to 2, and the total raw score is then mathematically converted to a 0 to 100 scale, with 0 representing the worst impact on quality of life and 100 representing no impact. A change of >8 points on the 0 to 100 scale is considered clinically meaningful. The composite score takes the raw scores from both subscales and converts them back to a single 0 to 100 scale.

[0188] Patients can also be assessed by the presence or absence of a Gorman grading system of diplopia. The Gorman assessment of subjective diplopia includes four categories: no diplopia (absent), diplopia when the patient is tired or awake (intermittent), diplopia on extreme gaze (non-constant), and continuous diplopia in straight-ahead or reading position (constant). Patients are scored according to what grade of diplopia they are experiencing. Improvement of greater than or equal to grade 1 is considered clinically meaningful.

[0189] In some embodiments, the method includes 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 in 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 3 weeks for at least 21 weeks.

[0190] 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 pharmacologic active compounds for the treatment of TAO.In some embodiments, the pharmaceutical composition further comprises a corticosteroid; rituximab or other anti-CD20 antibody; tricizumab or other anti-IL-6 antibody; or serine, infliximab or other TNF alpha antibody or thyroid stimulating hormone receptor (TSHR) inhibitor.

[0191] 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.

[0192] Also provided is a kit that is useful for carrying out the embodiments described herein. The kit includes a first container that contains or is packaged with the antibody described above. The kit also includes another container that contains or is packaged with a solution necessary or convenient for carrying out the embodiment. The container can be made of glass, plastic or foil, and can be a vial, bottle, pouch, tube, bag, etc. The kit can also include written information, such as a procedure for carrying out the embodiment, or analytical information, such as the amount of a reagent contained in the first container means. The container can also be in another container device, such as a box or bag, together with the written information.

[0193] Yet another embodiment defined herein is a kit for detecting IGF-1R protein in a biological sample.The kit comprises a container that holds one or more antibodies that bind to an epitope of IGF-1R protein, and instructions for using the antibodies to bind to IGF-1R protein to form an immunological complex, and to detect the formation of the immunological complex, the presence or absence of which correlates with the presence or absence of IGF-1R protein in the sample.An example of the container is a multi-well plate that allows simultaneous detection of IGF-1R protein in multiple samples.

[0194] In some embodiments, an antibody is provided that binds to IGF-1R protein. In some embodiments, the antibody is isolated. In some embodiments, the antibody specifically binds. In some embodiments, the antibody binds to correctly folded IGF-1R protein. In some embodiments, the antibody is specific to a particular IGF-1R conformational state (open or closed). In some embodiments, the antibody binds to IGF-1R protein in a cell membrane. In some embodiments, the antibody binds to IGF-1R protein in the cell membrane of an intact cell. 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. The 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 percent inhibition can be based on the function or activity of the protein in the absence of the antibody. In some embodiments, the antibody inhibits IGF-1R facilitated glucose transport, hi some embodiments, the antibody inhibits internalization of IGF-1R protein.

[0195] In some embodiments, the antibody comprises a sequence as defined herein, or an antigen-binding fragment thereof. In some embodiments, the antibody comprises a heavy chain CDR or an antigen-binding fragment thereof as described herein. The heavy chain can be one or more of the heavy chains described herein. In some embodiments, the antibody comprises a light chain or an antigen-binding fragment thereof as described herein.

[0196] In some embodiments, a method of treating, inhibiting or ameliorating an IGF-1R, associated pathology is provided. In some embodiments, the method comprises administering an antibody described herein or a pharmaceutical composition described herein to a subject to treat, inhibit or ameliorate an IGF-1R, associated pathology. In some embodiments, the pathology is as described herein.

[0197] In some embodiments, a method for detecting the presence or absence of IGF-1R in a sample is provided, comprising contacting the sample with one or more antibodies described herein, thereby detecting the binding of the antibody to an IGF-1R antigen. In some embodiments, the detection of binding indicates the presence of an IGF-1R antigen, or the absence of detection of binding to an IGF-1R antigen indicates the absence of an IGF-1R antigen. Detection can be performed in any known manner, for example, using a biosensor, ELISA, sandwich assay, etc. However, in some embodiments, the method comprises detecting the presence of a protein under non-denaturing conditions. Non-denaturing conditions can be used to detect a protein of interest in its native form or correctly folded form.

[0198] In some embodiments, a method is provided for identifying a test antibody that binds to an epitope on an IGF-1R protein, comprising contacting the test antibody with an epitope on an IGF-1R protein and determining whether the test antibody binds to the epitope.In some embodiments, the determining step comprises determining whether the test antibody binds to the protein and is competitively inhibited by an antibody comprising a sequence as provided herein.In some embodiments, the determining step comprises mutating one or more residues of the epitope or protein and determining the binding of the test antibody to the mutated epitope, and if the mutation reduces the binding of the test antibody compared to the non-mutated epitope, the test antibody is considered to bind to the epitope.

[0199] In some embodiments, a method is 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 as provided herein and detecting the presence of IGF-1R in the cell or on the surface of the cell. The difference 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 that modulates the internalization of IGF-1R protein. Thus, the antibody defined herein can be used to identify a test agent that modulates (increases or decreases) the internalization of IGF-1R protein. Test molecules that increase internalization, which will be measured as a decrease in binding of the anti-IGF-1R antibody to the IGF-1R protein on the cell surface, can be identified according to the methods provided herein. Test molecules that decrease internalization, which will be measured as an increase in binding of the anti-IGF-1R antibody to the IGF-1R protein on the cell surface, 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 surface expression can be measured by labeling the anti-IGF-1R antibody as defined herein.

[0200] In some embodiments, a method of treating thyroid eye disease in a subject in need thereof is provided. In some embodiments, the method comprises intravenously administering a dose of 10 mg / kg of an anti-IGF-1R antibody to the subject at regular intervals for a period sufficient to alleviate one or more symptoms associated with thyroid eye disease, the anti-IGF-1R antibody comprising a heavy chain comprising an HCDR1 of SEQ ID NO:7, an HCDR2 of SEQ ID NO:8, and an HCDR3 of SEQ ID NO:9, and a light chain comprising an LCDR1 of SEQ ID NO:4, an LCDR2 of SEQ ID NO:5, and an LCDR3 of SEQ ID NO:6. In some embodiments, the anti-IGF-1R antibody comprises a light chain and a heavy chain, the light chain comprising a variable region having an amino acid sequence of SEQ ID NO:2, and the heavy chain comprising a variable region having an amino acid sequence of SEQ ID NO:3. In some embodiments, the light chain comprises an amino acid sequence of SEQ ID NO:11. In some embodiments, the heavy chain comprises an amino acid sequence of SEQ ID NO:10. In some embodiments, the heavy chain comprises an amino acid sequence of SEQ ID NO:10, and the light chain comprises an amino acid sequence of SEQ ID NO:11. In some embodiments, the anti-IGF-1R antibody is administered by intravenous infusion. In some embodiments, the anti-IGF-1R antibody is administered every 3 weeks. In some embodiments, the anti-IGF-1R antibody is administered for a period sufficient for 5 doses. In some embodiments, the anti-IGF-1R antibody is administered for a period sufficient for 8 doses. In some embodiments, the anti-IGF-1R antibody is administered for a period selected from 3 weeks, 6 weeks, 9 weeks, 12 weeks, 15 weeks, 18 weeks, 21 weeks, 24 weeks or longer.

[0201] In some embodiments, an antibody is provided for use in a method of treating thyroid eye disease in a subject in need of such treatment.In some embodiments, an anti-IGF-1R antibody is provided for use in a method of treating thyroid eye disease in a subject in need of such treatment.

[0202] In some embodiments, the embodiments provided herein also include, but are not limited to, the following: 1. A method of treating thyroid eye disease in a subject in need of such treatment, comprising: administering a first dose of an antibody intravenously or subcutaneously to the subject, wherein the first dose is selected from the group consisting of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg; and administering one or more subsequent doses of the antibody intravenously or subcutaneously to the subject, wherein each subsequent dose is selected from the group consisting of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg. Including, the antibody comprises a heavy chain and a light chain, the heavy chain comprises an HCDR1 of SEQ ID NO:7, an HCDR2 of SEQ ID NO:8, and an HCDR3 of SEQ ID NO:9, and the light chain comprises an LCDR1 of SEQ ID NO:4, an LCDR2 of SEQ ID NO:5, and an LCDR3 of SEQ ID NO:6; or the antibody comprises a light chain comprising a variable region having the amino acid sequence of SEQ ID NO:2 and the heavy chain comprises a variable region sequence having the amino acid sequence of SEQ ID NO:3; or A method, wherein the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:11, and a heavy chain comprising the amino acid sequence of SEQ ID NO:10. 2. The method of claim 1, wherein the first dose is about 2 mg / kg. 3. The method of claim 1, wherein the first dose is about 3 mg / kg. 4. The method of claim 1, wherein the first dose is about 2.5 mg / kg. 5. The method of claim 1, wherein the first dose is about 5 mg / kg. 6. The method of claim 1, wherein the first dose is about 7.5 mg / kg. 7. The method of claim 1, wherein the first dose is about 10 mg / kg. 8. The method of claim 1, wherein the first dose is about 15 mg / kg. 9. The method of claim 1, wherein the first dose is about 20 mg / kg. 10. The method of any one of claims 2 to 9, wherein the amount of the one or more subsequent doses is the same as the amount of the first dose. 11. The method of any one of claims 2 to 9, wherein the amount of one or more subsequent doses is different from the amount of the first dose. 12. The method of any one of claims 2 to 11, wherein at least one of the one or more subsequent doses is about 2 mg / kg. 13. The method of any one of claims 2 to 11, wherein at least one of the one or more subsequent doses is about 3 mg / kg. 14. The method of any one of claims 2 to 11, wherein at least one of the one or more subsequent doses is about 5 mg / kg. 15. The method of any one of claims 2 to 11, wherein at least one of the one or more subsequent doses is about 7.5 mg / kg. 16. The method of any one of claims 2 to 11, wherein at least one of the one or more subsequent doses is about 10 mg / kg. 17. The method of any one of claims 2 to 11, wherein at least one of the one or more subsequent doses is about 15 mg / kg. 18. The method of any one of claims 2 to 11, wherein at least one of the one or more subsequent doses is about 20 mg / kg. 19. The method of any one of claims 1 to 18, wherein at least one of the one or more subsequent doses is administered 1, 2, 3, 4, 5, 6 or 8 weeks after the first dose. 20. The method of any one of claims 1 to 19, wherein only 1, 2, 3, 4, 5, 6 or 7 subsequent doses are administered to the subject. 21. The method of any one of claims 1 to 20, comprising administering a total of 2, 3, 4, 5, 6, 7 or 8 doses to the subject. 22. The method of any one of claims 1 to 21, wherein the subject's clinical activity score is reduced after two or three doses of the antibody. 23. The method of any one of claims 1 to 22, wherein each subsequent dose is administered 1, 2, 3, 4, 5, 6, 7, or 8 weeks after the previous dose. 24. The method of any one of claims 1 to 23, comprising administering at least one dose by intravenous infusion over 45 minutes to about 90 minutes or over 60 minutes to about 90 minutes. 25. The method of any one of claims 1 to 24, comprising administering at least one dose by subcutaneous administration. 26. The method of claim 25, wherein the subcutaneous administration is self-administration. 27. The method of any one of claims 1 to 26, further comprising administering to the subject one or more loading doses of the antibody prior to administering the first dose. 28. The method of any one of claims 1 to 26, further comprising administering to the subject a first loading dose of the antibody prior to administering the first dose, wherein the first loading dose is selected from the group consisting of about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 12.5 mg / kg, or about 12.5 mg / kg to about 15 mg / kg. 29. The method of claim 27 or 28, wherein a second loading dose of the antibody is administered to the subject after the first loading dose, and the first and second loading doses are administered before the first dose. 30. The method of claim 29, wherein the first loading dose and the second loading dose are the same dose amount. 31. The method of claim 29, wherein the first loading dose and the second loading dose are different dose amounts. 32. The method of any one of claims 27 to 31, wherein the first loading dose is about 5 mg / kg. 33. The method of any one of claims 27 to 31, wherein the first loading dose is about 7.5 mg / kg. 34. The method of any one of claims 27 to 31, wherein the first loading dose is about 10 mg / kg. 35. The method of any one of claims 27 to 31, wherein the first loading dose is about 12.5 mg / kg. 36. The method of any one of claims 27 to 31, wherein the first loading dose is about 15 mg / kg. 37. The method of any one of claims 27 to 36, wherein the second loading dose is about 5 mg / kg. 38. The method of any one of claims 27 to 36, wherein the second loading dose is about 7.5 mg / kg. 39. The method of any one of claims 27 to 36, wherein the second loading dose is about 10 mg / kg. 40. The method of any one of claims 27 to 36, wherein the second loading dose is about 12.5 mg / kg. 41. The method of any one of claims 27 to 36, wherein the second loading dose is about 15 mg / kg. 42. The method of any one of claims 27 to 41, wherein a first loading dose is administered to the subject 1, 2, 3, or 4 weeks before the first dose is administered. 43. The method of any one of claims 1 to 42, wherein the antibody is administered as part of a pharma- ceutically acceptable composition comprising the antibody and at least one pharma- ceutically acceptable excipient, and the antibody has a solubility in the pharma- ceutically acceptable composition of at least about 150 mg / ml. 44. The method of any one of claims 1-43, wherein the subject has had an unsatisfactory response to previous therapeutic agents for thyroid eye disease. 45. The method of claim 44, wherein the unsatisfactory response is one or more of the following: failure to reduce exophthalmos by 2 mm or more; failure to reduce CAS in one or more components or by 2 or more points; deterioration of 2 mm or more in the fellow eye; failure to reduce diplopia; failure to continue to improve diplopia for a period of time; failure to improve the Graves' Ophthalmopathy Quality of Life (GO-QoL) score by 8 or more points; and combinations thereof. 46. ​​A method of treating thyroid eye disease in a subject in need of such treatment, comprising: administering a first dose of an antibody intravenously or subcutaneously to the subject, wherein the first dose is selected from the group consisting of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg; and administering one or more subsequent doses of the antibody intravenously or subcutaneously to the subject, wherein each subsequent dose is selected from the group consisting of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg. and the antibody is as provided herein, including, but not limited to, the antibody comprises a heavy chain and a light chain, the heavy chain comprises an HCDR1 of SEQ ID NO:7, an HCDR2 of SEQ ID NO:8, and an HCDR3 of SEQ ID NO:9, and the light chain comprises an LCDR1 of SEQ ID NO:4, an LCDR2 of SEQ ID NO:5, and an LCDR3 of SEQ ID NO:6; or the antibody comprises a light chain comprising a variable region having the amino acid sequence of SEQ ID NO:2 and the heavy chain comprises a variable region sequence having the amino acid sequence of SEQ ID NO:3; or The antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:11 and a heavy chain comprising the amino acid sequence of SEQ ID NO:10. This includes: A method wherein one or more subsequent doses are administered when the subject has not responded adequately to one or more previous doses, as determined by clinical activity scores and / or exophthalmos measurements. 47. A method of improving treatment of thyroid eye disease in a subject who has previously received one or more treatments, comprising: The method includes administering to a subject intravenously or subcutaneously at least one dose of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg of an antibody, wherein the antibody is exemplified by an antibody as provided herein, e.g., the antibody comprises a heavy chain and a light chain, the heavy chain comprises an HCDR1 of SEQ ID NO:7, an HCDR2 of SEQ ID NO:8, and an HCDR3 of SEQ ID NO:9, and the light chain comprises an LCDR1 of SEQ ID NO:4, an LCDR2 of SEQ ID NO:5, and an LCDR3 of SEQ ID NO:6; or the antibody comprises a light chain comprising a variable region having the amino acid sequence of SEQ ID NO:2 and the heavy chain comprises a variable region sequence having the amino acid sequence of SEQ ID NO:3; or the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:11 and a heavy chain comprising the amino acid sequence of SEQ ID NO:10; A method wherein at least one dose results in an improvement in one or more measured values ​​as compared to the one or more measured values ​​prior to the at least one dose. 48. The method of claim 47, wherein the one or more measurements are selected from exophthalmos, CAS, level of deterioration in the fellow eye, score on the GO-QoL, and combinations thereof. 49. The method of claim 47 or 48, wherein if the subject does not exhibit a satisfactory response after at least one dose, one or more subsequent doses of the antibody are administered to the subject, each dose being selected from the group consisting of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg. 50. The method of claim 49, wherein the one or more subsequent doses improve one or more of: exophthalmos, CAS, level of deterioration in the fellow eye, score on GO-QoL, and combinations thereof, compared to before the one or more subsequent doses. 51. A method of treating thyroid eye disease in a subject in need of such treatment, comprising: administering a first dose of an antibody to a subject intravenously or subcutaneously, said first dose being selected from the group consisting of about 250 mg, about 300 mg, about 350 mg, or about 400 mg; and administering one or more subsequent doses of the antibody intravenously or subcutaneously to the subject, wherein each subsequent dose is selected from the group consisting of about 250 mg, about 300 mg, about 350 mg, or about 400 mg. wherein the antibody is represented by SEQ ID NO:1. 52. The method of claim 51, wherein the first dose is about 250 mg. 53. The method of claim 52, wherein the first dose is about 300 mg. 54. The method of claim 52, wherein the first dose is about 350 mg. 55. The method of claim 52, wherein the first dose is about 400 mg. 56. The method of any one of claims 51 to 55, wherein the amount of the first dose and the amount of the one or more subsequent doses are the same dose amount. 57. The method of any one of claims 51 to 55, wherein the amount of the first dose and the amount of the one or more subsequent doses are different dose amounts. 58. The method of any one of claims 51 to 57, wherein at least one of the one or more subsequent doses is about 250 mg. 59. The method of any one of claims 51 to 57, wherein at least one of the one or more subsequent doses is about 300 mg. 60. The method of any one of claims 51 to 57, wherein at least one of the one or more subsequent doses is about 350 mg. 61. The method of any one of claims 51 to 57, wherein at least one of the one or more subsequent doses is about 400 mg. 62. The method of any one of claims 51 to 61, wherein at least one of the one or more subsequent doses is administered 1, 2, 3, 4, 5, 6, or 8 weeks after the first dose. 63. The method of any one of claims 51 to 62, wherein only 1, 2, 3, 4, 5, 6 or 7 subsequent doses are administered to the subject. 64. The method of any one of claims 51 to 63, comprising administering to the subject a total of 2, 3, 4, 5, 6, 7 or 8 doses. 65. The method of any one of claims 51 to 64, wherein the subject's clinical activity score is reduced after two or three doses of the antibody. 66. The method of any one of claims 51 to 65, wherein each subsequent dose is administered 1, 2, 3, 4, 5, 6, 7, or 8 weeks after the previous dose. 67. The method of any one of claims 51 to 66, comprising administering at least one dose by intravenous infusion over 45 minutes to about 90 minutes, or over 60 minutes to about 90 minutes. 68. The method of any one of claims 51 to 67, comprising administering at least one dose by subcutaneous administration. 69. The method of claim 68, wherein the subcutaneous administration is self-administration. 70. The method of any one of claims 51 to 69, further comprising administering to the subject one or more loading doses of the antibody prior to administering the first dose. 71. The method of any one of claims 51 to 69, further comprising administering to the subject a first loading dose of the antibody prior to administering the first dose, wherein the first loading dose is selected from the group consisting of about 250 mg, about 300 mg, about 350 mg, or about 400 mg. 72. The method of claim 70 or 71, wherein a second loading dose of the antibody is administered to the subject after the first loading dose, and the first and second loading doses are administered before the first dose. 73. The method of claim 72, wherein the first loading dose and the second loading dose are the same dose amount. 74. The method of claim 73, wherein the first loading dose and the second loading dose are different dose amounts. 75. The method of any one of claims 70 to 73, wherein the first loading dose is about 250 mg. 76. The method of any one of claims 70 to 73, wherein the first loading dose is about 300 mg. 77. The method of any one of claims 70 to 73, wherein the first loading dose is about 350 mg. 78. The method of any one of claims 70 to 73, wherein the first loading dose is about 400 mg. 79. The method of any one of claims 70-78, wherein the second loading dose is about 250 mg. 80. The method of any one of claims 70-78, wherein the second loading dose is about 300 mg. 81. The method of any one of claims 70-78, wherein the second loading dose is about 350 mg. 82. The method of any one of claims 70-78, wherein the second loading dose is about 400 mg. 83. The method of any one of claims 70-78, wherein a first loading dose is administered to the subject 1, 2, 3, or 4 weeks before the first dose is administered. 84. The method of any one of claims 51 to 83, wherein the antibody is administered as part of a pharma- ceutically acceptable composition comprising the antibody and at least one pharma- ceutically acceptable excipient, and the antibody has a solubility in the pharma-ceutically acceptable composition of at least about 150 mg / ml. 85. The method of any one of claims 51-84, wherein the subject has had an unsatisfactory response to previous therapeutic agents for thyroid eye disease. 86. The method of claim 85, wherein unsatisfactory response is selected from failure to reduce exophthalmos by 2 mm or more; failure to reduce CAS in one or more components or by 2 or more points; deterioration of 2 mm or more in the fellow eye; failure to reduce diplopia; failure to continue to improve diplopia for a period of time; failure to improve the Graves' Ophthalmopathy Quality of Life (GO-QoL) score by 8 or more points; and combinations thereof. 87. The method of any one of claims 27-45 or 70-86, wherein the first loading dose and the second loading dose are administered about 1, about 2, or about 3 weeks apart. 88. The method of claim 87, wherein the second loading dose is administered about 1, about 2, or about 3 weeks before the first dose. 89. A method of treating thyroid eye disease in a subject in need of such treatment, comprising: administering a first dose of an antibody intravenously or subcutaneously to the subject, wherein the first dose is selected from the group consisting of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg; and administering one or more subsequent doses of the antibody intravenously or subcutaneously to the subject, wherein each subsequent dose is selected from the group consisting of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg. Including, A method wherein the antibody comprises a heavy chain and a light chain, the heavy chain comprises an HCDR1 of SEQ ID NO: 7, an HCDR2 of SEQ ID NO: 8, and an HCDR3 of SEQ ID NO: 9, and the light chain comprises an LCDR1 of SEQ ID NO: 4, an LCDR2 of SEQ ID NO: 5, and an LCDR3 of SEQ ID NO: 6. 90. The method of embodiment 89, wherein the light chain comprises a variable region having the amino acid sequence of SEQ ID NO: 2, and the heavy chain comprises a variable region sequence having the amino acid sequence of SEQ ID NO: 3. 91. The method of embodiment 89 or 90, wherein the light chain comprises the amino acid sequence of SEQ ID NO:11. 92. The method of any one of embodiments 89-91, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:10. 93. The method of embodiment 89, wherein the first dose is about 10 mg / kg. 94. The method of embodiment 93, wherein the one or more subsequent doses are about 10 mg / kg. 95. The method of any one of embodiments 89-93, wherein the amount of the one or more subsequent doses is the same as the amount of the first dose. 96. The method of any one of embodiments 89-94, wherein the amount of the one or more subsequent doses is different from the amount of the first dose. 97. The method of any one of embodiments 89-96, wherein at least one of the one or more subsequent doses is administered 1, 2, 3, 4, 5, 6, or 8 weeks after the first dose. 98. The method of embodiment 94, wherein the one or more subsequent doses are administered three weeks after the first dose. 99. The method of embodiment 94, wherein the subsequent doses are administered every 3 weeks for 4, 5, 6, 7 or 8 cycles after the first dose. 100. The method of embodiment 94, wherein the subsequent doses are administered every 3 weeks after the first dose for 5 or 8 cycles. 101. The method of embodiment 94, comprising administering a total of 5, 6, 7, or 8 doses to the subject. 102. The method of any one of embodiments 89-101, wherein after the first dose of the antibody, the subject's clinical activity score is reduced. 103. The method of any one of embodiments 89-102, wherein after two doses of the antibody, the subject's clinical activity score is reduced. 104. The method of embodiments 89-103, wherein after one or more subsequent doses, the subject's clinical activity score is reduced within 6 weeks of the first dose. 105. The method of embodiments 89-103, wherein after one or more subsequent doses, the subject's clinical activity score is reduced within 3 weeks of the first one or more subsequent doses. 106. The method of any one of embodiments 89-105, wherein the antibody is administered by intravenous infusion over 45 minutes to about 90 minutes, or over 60 minutes to about 90 minutes. 107. The method of embodiment 89, wherein the first dose is about 10 mg / kg and the one or more subsequent doses are about 10 mg / kg. 108. The method of any one of embodiments 89-106, wherein the antibody is administered as part of a pharma- ceutically acceptable composition comprising the antibody and at least one pharma- ceutically acceptable excipient, and the pharmaceutical composition comprises the antibody at a concentration of 20 mg / mL to about 30 mg / mL. 109. The method of embodiment 108, wherein the pharmaceutical composition comprises the antibody at a concentration of about 25 mg / mL. 110. The method of any one of embodiments 89-109, wherein the exophthalmos of the treated subject is reduced by at least, or about, 1-4 mm. 111. The method of embodiment 110, wherein exophthalmos is reduced by at least, or about, 2-3 mm. 112. The method of embodiment 110 or 111, wherein exophthalmos is reduced within 3 weeks of the first dose. 113. The method of embodiment 110 or 111, wherein exophthalmos is reduced within 6 weeks of the first dose. 114. The method of any one of embodiments 89-113, wherein the treated subject has a reduction in diplopia. 115. The method of embodiment 114, wherein the diplopia is relieved within 3 or 6 weeks of the first dose. 116. The method of any one of embodiments 89-115, wherein the subject has an improvement in Clinical Activity Score (CAS) within 3 or 6 weeks. 117. The method of embodiment 37, wherein the CAS score has an improvement of at least -2, -3, or -4. 118. The method of any one of embodiments 89-117, wherein the subject has a reduction in proptosis and an improvement in CAS score within 3 weeks or 6 weeks of the first dose. 119. A method of treating thyroid eye disease in a subject in need of such treatment, comprising: administering a first dose of 10 mg / kg of the antibody intravenously to the subject. Including, A method wherein the antibody comprises a heavy chain and a light chain, the heavy chain comprises an HCDR1 of SEQ ID NO: 7, an HCDR2 of SEQ ID NO: 8, and an HCDR3 of SEQ ID NO: 9, and the light chain comprises an LCDR1 of SEQ ID NO: 4, an LCDR2 of SEQ ID NO: 5, and an LCDR3 of SEQ ID NO: 6. 120. The method of embodiment 119, wherein the light chain comprises a variable region having the amino acid sequence of SEQ ID NO:2, and the heavy chain comprises a variable region sequence having the amino acid sequence of SEQ ID NO:3. 121. The method of embodiment 119, wherein the light chain comprises the amino acid sequence of SEQ ID NO:11. 122. The method of embodiment 119, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:10. 123. The method of embodiment 119, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 10, and the light chain comprises the amino acid sequence of SEQ ID NO: 11. 124. The method of any one of embodiments 119-123, further comprising administering a subsequent dose of about 10 mg / kg. 125. The method of embodiment 124, wherein the subsequent dose is administered about 3 weeks after the first dose. 126. The method of any one of embodiments 119-125, further comprising the step of administering, after the first dose, subsequent doses of about 10 mg / kg every 3 weeks. 127. The method of embodiment 126, wherein the subsequent doses are administered every 3 weeks for a total of 4 subsequent doses. 128. The method of embodiment 126, wherein the subsequent doses are administered every 3 weeks for a total of 7 subsequent doses. 129. The method of any one of embodiments 119-128, wherein the subject's exophthalmos is reduced by at least, or about, 1-4 mm. 130. The method of embodiment 129, wherein exophthalmos is reduced by at least, or about, 2-3 mm. 131. The method of embodiment 129 or 130, wherein exophthalmos is reduced within 3 weeks of the first dose. 132. The method of embodiment 129 or 130, wherein exophthalmos is reduced within 6 weeks of the first dose. 133. The method of any one of embodiments 119-132, wherein the treated subject has a reduction in diplopia. 134. The method of embodiment 133, wherein the diplopia is relieved within 3 or 6 weeks of the first dose. 135. The method of any one of embodiments 119-134, wherein the subject has an improvement in Clinical Activity Score (CAS) within 3 weeks or 6 weeks. 136. The method of embodiment 135, wherein the CAS score has an improvement of at least -2, -3, or -4. 137. The method of any one of embodiments 119-136, wherein the subject has a reduction in proptosis and an improvement in CAS score within 3 weeks or 6 weeks of the first dose. 138. A method of treating thyroid eye disease in a subject in need of such treatment, comprising: administering a 10 mg / kg dose of an anti-IGF-1R antibody intravenously to the subject at regular intervals for a period sufficient to reduce one or more symptoms associated with thyroid eye disease. Including, A method, wherein the anti-IGF-1R antibody comprises a heavy chain comprising an HCDR1 of SEQ ID NO: 7, an HCDR2 of SEQ ID NO: 8, and an HCDR3 of SEQ ID NO: 9, and a light chain comprising an LCDR1 of SEQ ID NO: 4, an LCDR2 of SEQ ID NO: 5, and an LCDR3 of SEQ ID NO: 6. 139. The method of embodiment 138, wherein the anti-IGF-1R antibody comprises a light chain and a heavy chain, the light chain comprising a variable region having the amino acid sequence of SEQ ID NO:2, and the heavy chain comprising a variable region having the amino acid sequence of SEQ ID NO:3. 140. The method of embodiment 138, wherein the light chain comprises the amino acid sequence of SEQ ID NO:11. 141. The method of embodiment 138, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:10. 142. The method of embodiment 138, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 10, and the light chain comprises the amino acid sequence of SEQ ID NO: 11. 143. The method of any one of embodiments 138-142, wherein the anti-IGF-1R antibody is administered by intravenous infusion. 144. The method of any one of embodiments 138-143, wherein the anti-IGF-1R antibody is administered every three weeks. 145. The method of any one of embodiments 138-144, wherein the anti-IGF-1R antibody is administered for a period sufficient for five doses. 146. The method of any one of embodiments 138-144, wherein the anti-IGF-1R antibody is administered for a period sufficient for eight doses. 147. The method of any one of embodiments 138-144, wherein the anti-IGF-1R antibody is administered for a period selected from 3 weeks, 6 weeks, 9 weeks, 12 weeks, 15 weeks, 18 weeks, 21 weeks, 24 weeks or longer. 148. The method of any one of embodiments 138-147, wherein the subject's exophthalmos is reduced by at least, or about, 1-4 mm. 149. The method of embodiment 148, wherein exophthalmos is reduced by at least, or about, 2-3 mm. 150. The method of embodiment 148 or 149, wherein exophthalmos is reduced within 3 weeks of the first dose. 151. The method of embodiment 148 or 149, wherein exophthalmos is reduced within 6 weeks of the first dose. 152. The method of any one of embodiments 148-151, wherein the treated subject has a reduction in diplopia. 153. The method of embodiment 152, wherein the diplopia is alleviated within 3 or 6 weeks of the first dose. 154. The method of any one of embodiments 138-153, wherein the subject has an improvement in Clinical Activity Score (CAS) within 3 weeks or 6 weeks. 155. The method of embodiment 154, wherein the CAS score has an improvement of at least -2, -3, or -4. 156. The method of any one of embodiments 138-155, wherein the subject has a reduction in proptosis and an improvement in CAS score within 3 weeks or 6 weeks of the first dose. 157. An antibody for use in treating thyroid eye disease in a subject in need of such treatment, comprising a heavy chain and a light chain, wherein the heavy chain comprises an HCDR1 of SEQ ID NO:7, an HCDR2 of SEQ ID NO:8, and an HCDR3 of SEQ ID NO:9, and the light chain comprises an LCDR1 of SEQ ID NO:4, an LCDR2 of SEQ ID NO:5, and an LCDR3 of SEQ ID NO:6; the antibody is administered intravenously or subcutaneously to the subject as a first dose of the antibody, the first dose being selected from the group consisting of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg; The antibody is administered intravenously or subcutaneously to the subject as one or more subsequent doses of the antibody, each subsequent dose being selected from the group consisting of about 1 mg / kg to about 2 mg / kg, about 2 mg / kg to about 5 mg / kg, about 3 mg / kg to about 5 mg / kg, about 5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 15 mg / kg, or about 15 mg / kg to about 20 mg / kg of the antibody. 158. The antibody of embodiment 157, wherein the light chain comprises a variable region having the amino acid sequence of SEQ ID NO: 2, and the heavy chain comprises a variable region sequence having the amino acid sequence of SEQ ID NO: 3. 159. The antibody of embodiment 157 or 158, wherein the light chain comprises the amino acid sequence of SEQ ID NO:11. 160. The antibody of any one of embodiments 157 to 159, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 10. 161. The antibody of embodiment 157, wherein the first dose is about 10 mg / kg. 162. The antibody of embodiment 161, wherein the one or more subsequent doses are about 10 mg / kg. 163. The antibody of any one of embodiments 157-161, wherein the amount of the one or more subsequent doses is the same as the amount of the first dose. 164. The antibody of any one of embodiments 157-161, wherein the amount of one or more subsequent doses is different from the amount of the first dose. 165. The antibody of any one of embodiments 157-164, wherein at least one of the one or more subsequent doses is administered 1, 2, 3, 4, 5, 6, or 8 weeks after the first dose. 166. The antibody of embodiment 165, wherein the one or more subsequent doses are administered 3 weeks after the first dose. 167. The antibody of embodiment 165, wherein the subsequent doses are administered every 3 weeks for 4, 5, 6, 7 or 8 cycles after the first dose. 168. The antibody of embodiment 165, wherein the subsequent doses are administered every 3 weeks after the first dose for 5 or 8 cycles. 169. The antibody of embodiment 165, wherein the treatment comprises administering a total of 5, 6, 7, or 8 doses to the subject. 170. The antibody of any one of embodiments 157-169, wherein after the first dose of the antibody, the subject's clinical activity score is reduced. 171. The antibody of any one of embodiments 157-170, wherein the subject's clinical activity score is reduced after two doses of the antibody. 172. The antibody of embodiments 157-171, wherein after one or more subsequent doses, the subject's clinical activity score is reduced within 6 weeks of the first dose. 173. The antibody of embodiments 1-171, wherein after one or more subsequent doses, the subject's clinical activity score is reduced within 3 weeks of the first one or more subsequent doses. 174. The antibody of any one of embodiments 1-173, wherein the antibody is administered by intravenous infusion over 45 minutes to about 90 minutes, or over 60 minutes to about 90 minutes. 175. The antibody of embodiment 157, wherein the first dose is about 10 mg / kg and the one or more subsequent doses are about 10 mg / kg. 176. The antibody of any one of embodiments 157 to 19, which is administered as part of a pharma- ceutical acceptable composition comprising the antibody and at least one pharma- ceutical acceptable excipient, wherein the pharmaceutical composition comprises the antibody at a concentration of 20 mg / mL to about 30 mg / mL. 177. The antibody of embodiment 176, wherein the pharmaceutical composition comprises the antibody at a concentration of about 25 mg / mL. 178. The antibody of any one of embodiments 157-177, wherein the exophthalmos of the treated subject is reduced by at least, or about, 1-4 mm. 179. The antibody of embodiment 178, in which exophthalmos is reduced by at least, or about, 2 to 3 mm. 180. The antibody of embodiment 178 or 179, in which exophthalmos is reduced within 3 weeks of the first dose. 181. The antibody of embodiment 178 or 179, in which exophthalmos is reduced within 6 weeks of the first dose. 182. The antibody of any one of embodiments 157-181, wherein the treated subject has reduced diplopia. 183. The antibody of embodiment 182, wherein the diplopia is alleviated within 3 or 6 weeks of the first dose. 184. The antibody of any one of embodiments 157-183, wherein the subject has an improvement in Clinical Activity Score (CAS) within 3 or 6 weeks. 185. The antibody of embodiment 184, having an improvement in CAS score of at least -2, -3, or -4. 186. The antibody of any one of embodiments 157-185, wherein the subject has a reduction in exophthalmos and an improvement in CAS score within 3 weeks or 6 weeks of the first dose. 187. An anti-IGF-1R antibody for use in treating thyroid eye disease in a subject in need of such treatment, comprising a heavy chain comprising an HCDR1 of SEQ ID NO:7, an HCDR2 of SEQ ID NO:8, and an HCDR3 of SEQ ID NO:9, and a light chain comprising an LCDR1 of SEQ ID NO:4, an LCDR2 of SEQ ID NO:5, and an LCDR3 of SEQ ID NO:6; An anti-IGF-1R antibody, wherein the anti-IGF-1R antibody is administered intravenously to a subject at regular intervals at a dose of 10 mg / kg of the anti-IGF-1R antibody for a period sufficient to alleviate one or more symptoms associated with thyroid eye disease. 188. The antibody of embodiment 187, wherein the anti-IGF-1R antibody comprises a light chain and a heavy chain, the light chain comprising a variable region having the amino acid sequence of SEQ ID NO:2, and the heavy chain comprising a variable region having the amino acid sequence of SEQ ID NO:3. 189. The antibody of embodiment 187, wherein the light chain comprises the amino acid sequence of SEQ ID NO:11. 190. The antibody of embodiment 187, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO:10. 191. The antibody of embodiment 187, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 10, and the light chain comprises the amino acid sequence of SEQ ID NO: 11. 192. The antibody of any one of embodiments 187-191, wherein the anti-IGF-1R antibody is administered by intravenous infusion. 193. The antibody of any one of embodiments 187-192, wherein the anti-IGF-1R antibody is administered every three weeks. 194. The antibody of any one of embodiments 187-193, wherein the anti-IGF-1R antibody is administered for a period sufficient for 5 doses. 195. The antibody of any one of embodiments 187-193, wherein the anti-IGF-1R antibody is administered for a period sufficient for 8 doses. 196. The antibody of any one of embodiments 187-195, wherein the anti-IGF-1R antibody is administered for a period selected from 3 weeks, 6 weeks, 9 weeks, 12 weeks, 15 weeks, 18 weeks, 21 weeks, 24 weeks or longer. 197. The antibody of any one of embodiments 187-196, wherein the subject's exophthalmos is reduced by at least, or about, 1-4 mm. 198. The antibody of embodiment 197, wherein exophthalmos is reduced by at least, or about, 2-3 mm. 199. The antibody of embodiment 197 or 198, wherein exophthalmos is reduced within 3 weeks of the first dose. 200. The antibody of embodiment 197 or 198, wherein exophthalmos is reduced within 6 weeks of the first dose. 201. The antibody of any one of embodiments 187-200, wherein the treated subject has reduced diplopia. 202. The antibody of embodiment 201, wherein the diplopia is alleviated within 3 or 6 weeks of the first dose. 203. The antibody of any one of embodiments 187-202, wherein the subject has an improvement in Clinical Activity Score (CAS) within 3 or 6 weeks. 204. The antibody of embodiment 203, having an improvement in CAS score of at least -2, -3, or -4. 205. The antibody of any one of embodiments 187-204, wherein the subject has a reduction in exophthalmos and an improvement in CAS score within 3 weeks or 6 weeks of the first dose.

[0203] The subject matter will now be described in connection with the following examples.These examples are merely for illustrative purposes, and the claims should in no way be considered as being limited to these examples, but rather should be considered as encompassing any and all variations that become evident as a result of the teachings provided herein.Those skilled in the art will know various non-critical parameters that can be changed or modified to produce essentially similar results. EXAMPLES

[0204] Example 1 VRDN-5000 exhibits better binding and increased antagonism compared to teprotumumab In vitro cell-based assays were performed in two different IFG1R-expressing cell lines: A549 cells (FIGS. 1A and 1B) and human ocular choroidal fibroblast (HOCF) cells (FIGS. 1C and 1D).

[0205] In cell-based antibody binding assays, VRDN-5000, sometimes referred to as VRDN-001, consistently demonstrated increasing levels of binding (as measured by mean fluorescence intensity) with increasing antibody concentration compared to teprotumumab or IgG controls in both A549 cells (Figure 1A; filled circles: VRDN-001; triangles: teprotumumab; open circles: IgG) and HOCF cells (Figure 1C; filled circles = VRDN; triangles: teprotumumab; open circles: IgG).

[0206] Similarly, VRDN-5000 consistently demonstrated increased antagonism (as measured by mean fluorescence intensity) at increasing antibody concentrations compared to teprotumumab in both A549 cells (Figure 1B; filled circles: VRDN-001; triangles: teprotumumab) and HOCF cells (Figure 1D; filled circles = VRDN; triangles: teprotumumab).

[0207] VRDN-5000 binds more strongly to IGFR1 than teprotumumab at picomolar and nanomolar antibody concentrations and exerts a stronger antagonistic effect. Example 2 Treatment of patients with thyroid eye disease and clinical evaluation of IGF-1R antibodies for thyroid eye disease

[0208] Subjects will receive an infusion of VRDN-5000 as disclosed herein. The number of infusions will be individualized for each subject and based on the clinical judgment of the investigator. The day 1 visit will be within 14 days after the last visit of the previous study. 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 only for subjects who were exophthalmos non-responders in the previous study, and subjects who relapsed in the previous study did not participate in the follow-up period. The visit window during the follow-up period is ±7 days.

[0209] The treatment period will be 24 weeks (6 months), during which time 8 infusions of teprotumumab will be administered.

[0210] Subjects who are exophthalmos non-responders will be scheduled to participate in the 6-month follow-up period in this extension study; subjects who relapse during the lead-in study and are re-treated during this extension study will not participate in the follow-up period.

[0211] Efficacy assessments will be performed on both eyes at each evaluation 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 measures.

[0212] Efficacy will be assessed by exophthalmos (measured as the exophthalmos rating of the clinical scale of severity using the Hertel instrument for consistency of measurement), CAS (7-item scale), diplopia (measured as part of the clinical scale of severity) and clinical scale of severity (including a movement limitation rating).

[0213] Quality of life will be assessed using the GO-QoL questionnaire.

[0214] Safety will be assessed by monitoring for AEs and concomitant medication use, immunogenicity studies, physical and ophthalmic examinations, vital signs, clinical safety laboratory assessments (complete blood count, chemistry (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).

[0215] Exophthalmos assessments will be performed using a Hertel exophthalmometer for consistency of measurements, with the same Hertel instrument and the same observer used at each assessment throughout the entire study period (unless strictly unavoidable), and the same intercanthal distance (ICD) will be used on each occasion.

[0216] Exophthalmos is measured for each eye on day 1 and 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 are recorded on the Clinical Measure of Exophthalmos Severity eCRF.

[0217] The antibodies are found to be effective in treating thyroid eye disease and in improving quality of life as defined herein. Example 3 A multiple ascending dose safety and efficacy study of anti-IGF-1R in normal healthy volunteers and subjects with thyroid eye disease (Ph1 / 2 study)

[0218] Normal healthy volunteers (NHV) and subjects with thyroid eye disease (TED subjects) will be treated with VRDN-5000 to study the safety, tolerability, preliminary efficacy, pharmacokinetic (PK) and pharmacodynamic (PD) profiles.

[0219] NHV and subjects with TED are screened for eligibility before treatment begins using the inclusion and exclusion criteria for NHV and TED subjects. NHV and TED subjects are administered intravenous infusions of VRDN-5000 (SEQ ID NO: 71). Each subject receives two infusions at a dose of 3 mg / kg, 10 mg / kg or 20 mg / kg, with each infusion being 3 weeks apart. Each infusion is administered over 90 minutes. For a given subject, the amount of the first and second doses is the same (e.g., a subject receives a first dose of 3 mg / kg and a second dose of 3 mg / kg, 3 weeks apart, etc.). NHV subjects are monitored for 6 weeks after the first dose, and TED subjects are monitored for 6 months after the first dose.

[0220] VRDN-5000 is supplied as a solution of 25 mg / mL antibody in a 5.1 mL fill volume in a 6 mL clear single-dose glass vial with a rubber septum, aluminum seal and plastic cap. Storage is at 2-8°C or frozen at -20°C.

[0221] VRDN-5000 will be dosed at a range of 3 mg / kg to 20 mg / kg. All subjects (NHV and those with TED) will be monitored for safety, efficacy and other endpoints.

[0222] Safety endpoints include adverse events (AEs), serious adverse events (SAEs) and laboratory evaluations, which will be monitored and recorded throughout the duration of the study.

[0223] Primary efficacy endpoints included exophthalmos responder rates (i.e., reduction in exophthalmos from baseline ≥ 2 mm [Hertel] among the "study" [larger exophthalmos] eyes) at weeks 6 and 12.

[0224] Other endpoints include blood levels of VRDN-001, IGF-1 and ADA at various time points before and after injection, as well as change from baseline in orbital fat volume as determined by magnetic resonance imaging (MRI); change in extraocular muscles as determined by MRI; Clinical Activity Score (CAS); change in subjective diplopia score; change in objective assessment of ocular motility as measured in five primary gaze positions with prism deviation; and change in Graves' Orbitopathy-Quality of Life (GO-QoL) score.

[0225] Dosing will be at 21-day intervals. Up to 48 subjects will be enrolled in the multiple ascending dose study (12-16 NHV and 16-32 subjects with TED in the multiple ascending dose study).

[0226] Single-dose PK measurements are determined after the first of two infusions, and repeat-dose PK measurements are determined after the second dose. PK and PD determinations are performed in NHV subjects to minimize the number of study visits required for TED subjects. Preliminary efficacy data are collected from TED subjects at 6, 12, and 24 weeks after the first of two infusions. Safety and tolerability data are collected from both NHV and TED subjects after treatment with VRDN-001. All measurements for NHV and TED subjects are performed as described herein.

[0227] The study is a randomized, double-blind (sponsor excluded), placebo-controlled study. Subjects and site personnel will be blinded to treatment. Pharmacists who prepare the infusion bags will not be blinded to treatment and will be provided with a 250 mL bag of saline with or without VRDN-5000 according to an Interactive Web-Based Response System (IWRS). The 250 mL bag will be infused over 90 minutes.

[0228] Three dose levels are evaluated: 3 mg / kg ("low"), 10 mg / kg ("medium"), and 20 mg / kg ("high"). Each subject receives two doses, three weeks apart, with each dose administered by intravenous infusion. The amount of dose given to each subject does not vary between doses (e.g., a subject receives a first 3 mg / kg dose and a second 3 mg / kg dose, three weeks apart, etc.).

[0229] The low-dose cohort included 4 NHVs randomized 3:1 (VRDN-5000 vs. placebo), and the medium- and high-dose cohorts included 4 NHVs and 8 TED subjects randomized 3:1 (VRDN-5000 vs. placebo) in each group.

[0230] The two NHV subjects of the low dose cohort are treated and followed up until one week after their first infusion, after which the remaining two subjects of the second cohort are treated. Dose-limiting toxicity (DLT) is a drug-related safety event of a severity that requires discontinuation of treatment and / or prevents dose escalation of VRDN-001. If a subject experiences a DLT, an additional four NHV subjects are enrolled, and escalation to the next dose is performed only if no further subjects experience a DLT.

[0231] The fourth NHV subject in the low dose cohort will begin escalation to the mid dose level one week after receiving their second infusion, following review of safety data by the Data Safety Monitoring Board (DSMB). Two NHV subjects will then be enrolled in the mid dose cohort and followed until one week after their first infusion, after which additional NHV and TED subjects will be enrolled at that dose level.

[0232] Escalation to the high dose level will occur once the fourth NHV subject in the medium dose cohort has been followed for one week after their second infusion, and if no more than one subject at that dose has experienced a DLT. This will occur after review of safety data by the DSMB. If both the medium and high dose cohorts show similar evidence of clinical activity in terms of exophthalmos response rates, eight TED subjects will be enrolled at the low dose (3.0 mg / kg) and a further cohort of 12 subjects (4 NHV and 8 TED subjects) will be enrolled at the medium dose (5.0 mg / kg) to generate a dose-response curve for clinical activity.

[0233] Study procedures for NHV are as follows: All NHV subjects are screened within 28 days prior to treatment and undergo a physical examination and ECG to rule out any abnormalities that would preclude study participation. Subjects are admitted to the Clinical Pharmacology site no later than 24 hours prior to each infusion and remain for 7 days after each of the two infusions for PK sample collection. Subjects' vital signs and ECG (telemetry) are continuously monitored during the infusion and skin injection sites are regularly tested for local tolerance. Subjects return to the site at designated times for further blood sampling and evaluations as outlined below. Web-based, physician-supervised hearing tests are performed prior to and 3 weeks after each infusion.

[0234] Blood samples for PK analysis and measurement of IGF-1 levels are collected by an indwelling venous catheter inserted in the forearm opposite the infusion arm. PK samples are collected before the start of each infusion, 5 minutes before the end of the infusion, and 2, 4, 8, and 12 hours after the infusion, with additional samples collected 1, 3, 7, 14, and 21 days after each infusion, and a final sample collected 28 days after the second infusion. Blood samples for IGF-1 levels are collected before each infusion, and 1, 2, 3, 7, 14, and 21 days after each infusion. Additional blood samples are collected for measurement of anti-drug antibodies (ADA) before each infusion of VRDN-001, and again 21 days after each infusion. Fasting blood and urine samples are collected for hematology, chemistry, and coagulation parameters and standard urinalysis at screening, immediately before each infusion, and 7 days after each infusion. NHVs undergo a full body examination and ECG at their 7-week visit.

[0235] The study procedures for TED subjects are as follows: TED subjects are screened for eligibility, medical history, and duration of TED 28 days prior to inclusion in the study. On the day prior to each injection, subjects undergo exophthalmos measurement, CAS assessment, diplopia score assessment, prism measurement of eye movements in five gaze positions, completion of the GO-QoL questionnaire, fundus examination, biomicroscopy, intraocular pressure (IOP), and hearing testing. These assessments are repeated at the follow-up visits on days 43 and 85 (weeks 6 and 12). Subjects undergo a full body examination and have an ECG performed on them, which is recorded at screening and repeated at the week 6 follow-up visit. Orbital MRI is performed within 3 days prior to both injections and repeated within (±) 3 days either side of the week 6 and 12 visits. Facial photography is performed at screening and at the week 12 and 24 visits. Injections are administered to TED subjects at the injection clinic, and subjects' vital signs and ECG are continuously monitored during the injections. Skin injection sites will be regularly inspected for local tolerance. Study site personnel will call TED subjects the day after each injection to ensure their physical and mental well-being and to inquire if any AEs have occurred since their discharge from the injection clinic the previous day. Subjects will be instructed to call the study site if they have any health concerns, and additional study visits will be arranged at the request of either the PI or the subject. All ocular evaluations will be performed on both eyes. These will be performed immediately prior to each injection on days 1 and 21, and again 3 weeks after the second injection. Follow-up visits for evaluation of exophthalmos will be performed 12 and 24 weeks after the first injection. Web-based, physician-supervised hearing tests will be performed before and 3 weeks after each injection.

[0236] Blood samples for PK and IGF-1 levels are taken by an indwelling venous catheter inserted in the forearm opposite to the injecting arm before starting the first infusion, 5 minutes before the end of the infusion, and 2 and 4 hours after the infusion, and are repeated at the same time points for the second infusion. Further samples are taken at each visit on days 43 and 50. Additional blood samples are taken before each infusion for the measurement of ADA, and again 3 weeks after the second infusion. Fasting blood and urine samples are taken for hematology, chemistry and coagulation parameters and standard urinalysis at screening, 3 weeks after the first infusion (the day before the second infusion), and 3 weeks after the second infusion.

[0237] The antibodies are found to be safe in NHVs, and are both safe and effective in treating thyroid eye disease and improving quality of life as defined herein in subjects with TED. Example 4 Extension Study of Anti-IGF-1R in Thyroid Eye Disease and in Subjects with Thyroid Eye Disease

[0238] Subjects with thyroid eye disease (TED subjects) will be treated with VRDN-5000 in an extension study following completion of a prior study for safety, tolerability, preliminary efficacy, pharmacokinetic (PK) and pharmacodynamic (PD) profiles.

[0239] TED subjects are administered an intravenous infusion of VRDN-5000. Each subject receives two infusions at a dose of 3 mg / kg, 10 mg / kg, or 20 mg / kg, with each infusion separated by 3 weeks. Each infusion is administered over 90 minutes. For a given subject, the first and second doses are the same (e.g., first and second doses of 3 mg / kg; first and second doses of 10 mg / kg; or first and second doses of 20 mg / kg). TED subjects are monitored for 6 months after the first dose.

[0240] VRDN-5000 is supplied as a solution of 25 mg / mL antibody in a 5.1 mL fill volume in a 6 mL clear single-dose glass vial with a rubber septum, aluminum seal and plastic cap. Storage is at 2-8°C or frozen at -20°C.

[0241] VRDN-5000 will be dosed at doses ranging from 3 mg / kg to 20 mg / kg. All subjects will be monitored for safety, efficacy and other endpoints.

[0242] Safety endpoints include adverse events (AEs), serious adverse events (SAEs) and laboratory evaluations, which will be monitored and recorded throughout the duration of the study.

[0243] Primary efficacy endpoints included the rate of exophthalmos responders (i.e., reduction in exophthalmos from baseline ≥ 2 mm [Hertel] among the "study" [larger exophthalmos] eyes) at 24 weeks.

[0244] Other endpoints include blood levels of VRDN-001, IGF-1 and ADA at various time points before and after injection, as well as change from baseline in orbital fat volume as determined by magnetic resonance imaging (MRI); change in extraocular muscles as determined by MRI; Clinical Activity Score (CAS); change in subjective diplopia score; change in objective assessment of ocular motility as measured in five primary gaze positions with prism deviation; and change in Graves' Orbitopathy-Quality of Life (GO-QoL) score.

[0245] Dosing will occur at 21-day intervals. Up to 48 subjects will participate in the extension study. The total number of subjects will depend on the results from the repeat dose escalation study in Example 3. If all 48 subjects are enrolled, the extension study will include 3 randomized (1:1:1; 16 per cohort / arm) in a double-blind (with sponsor) placebo-controlled design comparing 2 active treatment arms (4 vs. 8 infusions) to a placebo arm.

[0246] Assuming a 50% difference in exophthalmos reduction for each active arm versus placebo, the extension study has 80% power to test each dose regimen versus placebo. To this end, a one-sided type I error level of 0.025 is divided equally between both comparisons (Bonferroni correction), resulting in a pairwise type I error level of 0.0125.

[0247] The extension study will begin following completion of the multiple dose ascending study described in Example 3. This study will continue to investigate the clinical activity of VRDN-5000 at the lowest dose that demonstrates a clinically meaningful efficacy signal in the multiple dose ascending study of Example 3 and will address dose response exploration.

[0248] The specific parameters for dose and regimen in the extension cohorts will be data driven, including PK data determined therein, from the repeat dose escalation study in Example 3. If the doses tested in the repeat dose escalation study in Example 3 show comparable efficacy signals, additional lower dose cohorts will be explored. If not, one extension cohort will compare 4 infusions with another extension cohort receiving 8 infusions at the selected dose, and the defined treatment duration in all extension cohorts will be fully supported by toxicology data available at the time of the first infusion.

[0249] Subjects in all three cohorts receive the same number of injections to maintain blinding. Ophthalmic evaluations are performed on both eyes, and subjects undergo the following assessments performed: exophthalmos, CAS assessment, diplopia score assessment, prism measurement of eye movements in five gaze positions, completion of the GO-QoL questionnaire, fundus examination, biomicroscopy, and IOP. These assessments are performed the day before each injection and repeated at the 24 and 52 week visits. Web-based, physician-supervised hearing tests are performed one day before each injection and again at the 24 week visit. Orbital MRI is performed within 3 days before the first injection and repeated within (±) 3 days of the 12, 24, and 52 week visits. Facial photography is performed at screening and at the 24 and 52 week visits. Injections are administered to TED subjects at the injection clinic, and subjects' vital signs and ECG are continuously monitored during the injection. Skin injection sites are regularly inspected for local tolerance.

[0250] Blood samples for PK and IGF-1 levels are taken via an indwelling venous catheter inserted in the forearm opposite the injecting arm before the start of the first infusion, 5 minutes before the end of the infusion, and 2 and 4 hours after the infusion, with additional samples taken 1, 3, 7, 14, and 21 days after the infusion. These sample collection times are repeated immediately before and after the fourth infusion. A single sample is taken before each infusion at weeks 3, 6, 12, 15, 18, and 21. Additional samples are taken at weeks 24, 25, and 52. Subjects are offered the option of having a blood sample taken at their home or workplace by a phlebotomist / nurse if visiting the study site proves inconvenient due to other commitments. Additional blood samples are taken for measurement of ADA before each infusion and again at the week 24, 25, and 52 visits.

[0251] Fasting blood and urine samples will be collected for hematology, chemistry, and coagulation parameters and standard urinalysis at screening, and again 3 weeks after the 4th (visit the day before the 12th week infusion) and 8th infusion (week 24). Study site personnel will call TED subjects the day after each infusion to ensure their physical and mental well-being and to ask if any AEs have occurred since their discharge from the infusion clinic the day before. Subjects will be evaluated for any AEs at all study visits, and subjects will be instructed to call the study site if they have any health concerns at any time during the study. Additional study visits will be coordinated at the request of either the PI or the subject. The DSMB will review safety and laboratory data at 6-month intervals during the study.

[0252] The antibodies are found to be effective in treating thyroid eye disease and improving quality of life as defined herein in subjects with TED.

[0253] Example 5 VRDN-5000 is an antagonist antibody against insulin-like growth factor-1 receptor (IGF-1R) in development for the treatment of thyroid eye disease (TED). TED is caused 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 fibroblasts to the orbital tissue where they mediate the deposition of hyaluronic acid and the expansion of orbital muscle and fat. IGF-1R antagonism has been shown to reverse this orbital tissue expansion and provide robust relief of symptoms in TED patients.

[0254] VRDN-5000 is a humanized monoclonal antibody that targets IGF-1R. The IGF-1R binding and antagonist properties of VRDN-5000 were analyzed.

[0255] method

[0256] Surface plasmon resonance (SPR): Antibodies were captured by immobilized anti-Fc and recombinant IGF-1R extracellular domain (ECD) was run as analyte. Association and dissociation rate constants (ka and kd, respectively), as well as the equilibrium dissociation constant KD, were derived by global fitting of the data to a single-site model.

[0257] Epitope binning: VRDN-5000 was immobilized on the chip surface by amine coupling and used to capture IGF-1R-ECD, after which teprotumumab was flowed over the chip.

[0258] Cell binding: A549 human lung adenocarcinoma cells or primary human ocular choroidal fibroblasts (HOCF) were incubated with various concentrations of VRDN-5000 or teprotumumab. A single dose 50 nM 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 on live cells. Median fluorescence intensity (MFI) of viable 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).

[0259] Internalization: Cells were incubated with various concentrations of the antibody of interest for 60 min at 4° C. and 37° C. Cells were then washed three times and incubated with FITC-labeled goat anti-human Fc secondary antibody for 30 min at 4° C. The MFI of viable cells was measured by flow cytometry and data was analyzed using FlowJo software.

[0260] Cell surface marker expression: HOCF cells were incubated with 10ug / mL of directly labeled antibodies or IgG isotype control. Median fluorescence intensity (MFI) was measured by flow cytometry and data was analyzed using FlowJo software.

[0261] Antagonism: Serum-starved A549 or HOCF cells were preincubated with various concentrations of test antibodies for 1 h at 37°C and then stimulated by addition of 100 ng / mL (A549) or 200 ng / mL (HOCF) IGF-1 for 7 min at 37°C. Phosphorylated IGF-1R (pIGF1R) in biological duplicates 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-variable slope(four parameters)).

[0262] result

[0263] VRDN-5000 binds to IGF-1R with sub-nanomolar affinity. The bottom figure, Figure 2A, shows that increasing concentrations of IGF-1R-ECD bound to anti-FC capture VRDN-5000 or teprotumumab manifest a stepwise increase in SPR signal, allowing a global fit to the binding model. After washout of IGF-1R, VRDN-5000 shows a more persistent binding interaction. Figure 2B shows that IGF-1R-ECD bound strongly to immobilized VRDN-5000. Teprotumumab showed no binding to the IGF-1R:VRDN-5000 complex, suggesting that teprotumumab and VRDN-5000 have no epitope overlap.

[0264] VRDN-5000 binds to IGF-1R on A549 cells with high affinity. As shown in Figure 3A-C, binding of VRDN-5000 to A549 cells was assessed by flow cytometry and found to have a similar binding distribution as teprotumumab at three different concentrations. As shown in Figure 3D, the binding dose-response curve demonstrated a VRDN-5000 EC50=0.1 nM. As shown in Figure 3E, VRDN-5000, VRDN-2700 (VRDN-5000 with M252Y, S254T and T256E mutations in the Fc domain), and teprotumumab show comparable binding at temperatures that block IGF-1R receptor internalization. FIG. 3F shows that VRDN-5000, VRDN-2700, which has M252Y, S254T, and T256E mutations in the Fc domain, and teprotumumab cause comparable levels of internalization (approximately 50%) as measured by reduced membrane IGF-1R receptor levels at 37° C. versus 4° C.

[0265] HOCF as an in vitro model of TED pathology

[0266] CD34+, Thy-1+ orbital fibroblasts are involved in extracellular matrix deposition and pathogenic fibrosis in TED5. Here, we show that HOFCs express (A) IGF-1R and (B) TSHR, as well as (C) CD34 and Thy-1. This demonstrates that they can be used as an in vitro model system for IGF-1R function in TED5. Data are shown in Figure 4A-C.

[0267] VRDN-5000 binds with high affinity to IGF-1R on HOCF cells.

[0268] Figures 5A-B show the binding of VRDN-5000 to HOCF cells, which was assessed by flow cytometry and found to have approximately the same binding as teprotumumab at three different concentrations. Panel D shows a binding dose response curve that demonstrated VRDN-5000 with an EC50=0.4 nM.

[0269] VRDN-5000 is a subnanomolar IGF-1R agonist. VRDN-5000 potently inhibits IGF-1 stimulated receptor phosphorylation on A549 cells (IC50=0.09 nM) and HOCF cells (IC50=0.09 nM), as shown in Figure 6A-B.

[0270] Example 6 VRDN-5000 is a more potent inhibitor of IGF-1 binding to IGF1R compared to teprotumumab.

[0271] The binding of IGF-1 to IGF-1R present on the surface of cells was determined. Briefly, labeled IGF1 was incubated with cells in the presence of VRDN-5000 or teprotumumab, or in the absence of antibody (negative control). Cells were washed, and then the IGF-1 bound to cells was determined by detecting the presence of IGF1 label. As shown in Figure 7, VRDN-5000 was found to be a more potent inhibitor. Maximum inhibition was found to be 94% for VRDN-5000 and 48% for teprotumumab.

[0272] Inhibition was also evaluated for IGF-1-induced IGF1-R phosphorylation. Cell cultures were preincubated with antibodies (VRDN-5000 or teprotumumab) and IGF1 stimulants. Cells were lysed and pIGF1R was measured. VRDN-5000 was found to have a maximal inhibition of autophosphorylation of 96%, while teprotumumab was found to have a maximal inhibition of autophosphorylation of 76%. These results are shown in Figure 8.

[0273] A more potent inhibition of VRDN-5000 on IGF1 activity was also found by measuring Akt phosphorylation. Briefly, cell cultures were incubated with antibodies (VRDN-5000 or teprotumumab) and stimulated with IGF1. Cells were lysed and pAKT was measured using a standard assay. VRDN-5000 was found to have a maximal inhibition of Akt phosphorylation of 93%, while teprotumumab was found to have a maximal inhibition of Akt phosphorylation of 66%.

[0274] These results demonstrate that the VRDN-5000 and teprotumumab epitopes on IGF-1R overlap, that VRDN-5000 binds to IGF-1R on cells with a subnanomolar EC50, that VRDN-5000 promotes IGF-1R internalization, and that VRDN-5000 inhibits IGF-1R phosphorylation with a subnanomolar IC50. Thus, VRDN-5000 binds, antagonizes, and internalizes IGF-1R at subnanomolar concentrations. This suggests that VRDN-5000 should be used for potential, potent inhibition of the pathophysiological functions that cause TED.

[0275] Example 7 VRDN-5000 treats thyroid eye disease in subjects, with reduction in exophthalmos occurring within 3 weeks of the first dose. Two injections of VRDN-5000 at a dose of 10 mg / kg, spaced 3 weeks apart, resulted in rapid and significant improvement in exophthalmos, CAS, and diplopia at week 6. Results demonstrate that at week 6, patients treated with VRDN-5000 have an exophthalmos response: 5 / 6 patients (83%); median time to exophthalmos response: 3 weeks; CAS response: 6 / 6 (100%) CAS score 0 or 1: 4 / 6 (67%); overall response: 5 / 6 patients (83%); and diplopia resolution: 3 / 4 (75%). The mean reduction in exophthalmos and improvement in diplopia observed as soon as 6 weeks after the first injection of VRDN-5000 was significantly faster than that published for teprotumumab (Smith et al., Teprotumumab for Thyroid-Associated Ophthalmopathy, N Engl J Med 2017;376:1748-61; Douglas et al., Teprotumumab for the Treatment of Active Thyroid Eye Disease, N. Engl J Med 2020;382:341-52; and Douglas et al., Teprotumumab Efficacy, Safety, and Durability in Longer-Duration Thyroid Eye Disease and Re-treatment, Ophthalmology 2022, vol 129, no. 4). Comparative data are shown in Figures 9-14B.

[0276] As the data demonstrate, on average, subjects had a reduction or improvement in at least two of the following: exophthalmos, diplopia, and CAS scores, which was not observed in the placebo cohort. These data demonstrate an unexpected result of how quickly VRDN-5000 can achieve a therapeutic effect that could not have been predicted.

[0277] 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 was specifically and individually indicated to be incorporated by reference. Applicants intend, in accordance with 37 CFR §1.57(b)(1), that this statement of incorporation by reference relates to each and every individual publication, database entry (e.g., Genbank sequence or GeneID entry), patent application, or patent, each of which is clearly identified in accordance with 37 CFR §1.57(b)(2), even if such citation is not immediately adjacent to the dedicated statement of incorporation by reference. The incorporation of a dedicated statement of incorporation by reference, if any, in this specification does not in any way weaken this general statement of incorporation by reference. Citation of any references herein is not intended as an admission that the references are pertinent prior art, nor does it constitute any admission as to the contents or date of these publications or documents.

[0278] 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 and accompanying figures. Such modifications are intended to be within the scope of the embodiments and any accompanying claims.

[0279] 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 fall within the scope of the disclosure and any appended claims.

Claims

1. 1. A pharmaceutical composition for use in treating thyroid eye disease in a subject in need thereof, comprising: comprising an antibody, the antibody is administered at 10 mg / kg as a first dose; and the antibody is administered at 10 mg / kg in one or more subsequent doses; A pharmaceutical composition wherein the antibody comprises a heavy chain and a light chain, the heavy chain comprising an HCDR1 of SEQ ID NO: 7, an HCDR2 of SEQ ID NO: 8, and an HCDR3 of SEQ ID NO: 9, and the light chain comprising an LCDR1 of SEQ ID NO: 4, an LCDR2 of SEQ ID NO: 5, and an LCDR3 of SEQ ID NO:

6.

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

3.

3. 3. The pharmaceutical composition of claim 1 or 2, wherein the light chain comprises the amino acid sequence of SEQ ID NO: 11 and the heavy chain comprises the amino acid sequence of SEQ ID NO:

10.

4. 10. The pharmaceutical composition of claim 1, wherein the one or more subsequent doses are administered three weeks after the first dose.

5. The pharmaceutical composition of claim 1, wherein the antibody is administered by intravenous infusion.

6. 6. The pharmaceutical composition of claim 5, wherein the antibody is administered by intravenous infusion over a period of from 45 minutes to about 90 minutes, or from 60 minutes to about 90 minutes.

7. 10. The pharmaceutical composition of claim 1, wherein the subject's exophthalmos is reduced within three weeks of the first dose.

8. 10. The pharmaceutical composition of claim 1, wherein the subject's diplopia is alleviated within 3 or 6 weeks of the first dose.

9. 10. The pharmaceutical composition of claim 1, wherein the subject has an improvement in Clinical Activity Score (CAS) within 3 or 6 weeks.

10. 10. The pharmaceutical composition of claim 1, wherein the subject has a reduction in proptosis and an improvement in CAS score within 3 weeks or 6 weeks of the first dose.

Images