A method of treating an inflammatory condition of the eye using an IGF-1R ligand conjugated to a disease modifier

Abstract

The present invention provides a method for treating an inflammatory condition of the eye in a subject, the method comprising administering to the subject an effective amount of a conjugate comprising an IGF-1R ligand, or a portion or variant thereof, and a disease modifying agent.

Description

Technical Field

[0001] (Cross - Reference to Related Applications) This application claims the benefit of U.S. Provisional Patent Application No. 63 / 367,238, filed Jun. 29, 2022.

[0002] The subject matter of the present disclosure generally relates to a method of treating ophthalmic inflammatory conditions, particularly by administering to a subject an insulin - like growth factor 1 receptor (IGF - 1R) ligand conjugated to a disease - modifying agent.

[0003] (Reference to Sequence Listing) The sequence listing, which is contained in the file named 597948SEQLIST.xml, is 17 kilobytes in size, was created on Jun. 29, 2023, and is hereby incorporated by reference into this specification.

Background Art

[0004] Ophthalmic inflammatory conditions encompass a wide range of conditions, from easily treatable states to severe vision - threatening emergencies that can lead to permanent vision loss. Inflammation can occur in response to infection, allergy, irritation, injury, or trauma to the eye or surrounding tissues. Additionally, many systemic autoimmune diseases have ocular manifestations.

[0005] One example is thyroid eye disease (TED), an autoimmune disease characterized by progressive inflammation and damage to the tissues surrounding the eye. TED occurs in approximately 40% of patients with Graves' disease and is estimated to affect 16 in 100,000 women and 2.9 in 100,000 men, with men more frequently having the severe form of the disease. Pathological changes involve enlargement of the extraocular muscles and increased orbital fat, leading to proptosis, which causes orbital congestion, periorbital edema, and other symptoms that can sometimes be severely vision - threatening. Other symptoms include strabismus, diplopia, limitation of eye movement, eyelid retraction, pain, astigmatism, facial deformity, and decreased quality of life.

[0006] In this technical field, there remains a need for improved methods of treating TED and other inflammatory conditions of the eye. SUMMARY OF THE INVENTION

[0007] According to this specification, the present application describes a method of treatment using an IGF-1R ligand conjugated to a disease-modifying agent.

[0008] In one embodiment, the present application provides a method for treating an inflammatory condition of the eye in a subject in need thereof, the method comprising administering to the subject an effective amount of a conjugate, the conjugate comprising an IGF-1R ligand and a disease-modifying agent.

[0009] In some embodiments, the inflammatory condition of the eye is TED, uveitis, scleritis, keratitis, or conjunctivitis.

[0010] In some embodiments, the inflammatory condition of the eye is an orbital inflammatory disease. In some embodiments, the orbital inflammatory disease is idiopathic orbital inflammation, orbital inflammatory pseudotumor, orbital myositis, inflammatory orbital cellulitis, optic nerve sheath meningitis, episcleritis, diffuse orbital inflammation, orbital apex syndrome, or sclerosing orbital inflammation.

[0011] In some embodiments, the inflammatory condition of the eye is TED.

[0012] In some embodiments, the TED is acute TED, inactive TED, chronic TED, moderate to severe TED, or vision-threatening (super-severe) TED.

[0013] In some embodiments, the treatment of TED using the conjugates of the present invention results in a reduction of proptosis in the subject. In certain embodiments, the reduction of proptosis is reduced by at least 2 mm, at least 3 mm, or at least 4 mm. In another embodiment, the reduction of proptosis is evaluated by a proptometer or orbital imaging. In certain embodiments, the orbital imaging is computed tomography scan or magnetic resonance imaging. In some embodiments, the reduction of proptosis is related to the reduction of extraocular muscle volume and / or orbital fat volume.

[0014] In some embodiments, the treatment of TED using the conjugates of the present invention results in a reduction of the clinical activity score (CAS) in the subject. In certain embodiments, the CAS is reduced by at least 2 points or at least 3 points. In another specific embodiment, the CAS is reduced to 1 or reduced to 0.

[0015] In some embodiments, the treatment of TED using the conjugates of the present invention results in a reduction of the severity of diplopia in the subject. In another embodiment, the reduction of the severity of diplopia is measured by the Gorman subjective diplopia score. In some embodiments, the diplopia is constant diplopia, intermittent diplopia, or indeterminate diplopia. In some embodiments, the treatment results in complete remission of diplopia. In another embodiment, the severity of diplopia in the subject is reduced by at least 1 grade.

[0016] In some embodiments, the treatment of TED using the conjugates of the present invention results in an improvement in the quality of life of the subject. In another embodiment, the improvement in the quality of life is measured by the Graves' ophthalmopathy quality of life (GO-QoL) assessment scale, or the visual function subscale of GO-QoL, or the appearance subscale of GO-QoL. In certain embodiments, the quality of life is improved by at least 8 points on the GO-QoL scale, or the visual function subscale of GO-QoL, or the appearance subscale of GO-QoL.

[0017] In some embodiments, the treatment results in an improvement in neuropathy of the optic nerve.

[0018] In some embodiments, the treatment results in a reduction in retro-orbital edema.

[0019] In some embodiments, the treatment results in an improvement in monocular retraction movement. In certain embodiments, the improvement in monocular retraction movement is measured by a light reflex test. In another specific embodiment, the improvement in monocular retraction movement is at least 10 degrees.

[0020] In some embodiments, the treatment results in a change or inhibition of the function of IGF-1R-expressing cells in the subject. In other embodiments, the treatment results in a reduction in the number of IGF-1R-expressing cells in the subject. In certain embodiments, the reduction is caused by killing of IGF-1R-expressing cells. Without being bound by theory, simple inhibition of the IGF-1R pathway may continue to act as a disease effector, or may be upregulated, resulting in compensatory mechanisms or pathways that can lead to adaptive resistance, and thus may be insufficient as a completely curative treatment due to other potentially redundant signaling resulting from such compensatory mechanisms or pathways. Therefore, targeted delivery of a disease-modifying agent, including an agent that results in killing of IGF-1R-expressing cells, may provide benefits that exceed (including being preferential to) simple IGF-1R pathway inhibition in order to bypass the effects of such potentially redundant pathways and / or compensatory pathways.

[0021] In another specific embodiment, the IGF-1R-expressing cells are orbital fibroblasts (OF). In another embodiment, the treatment results in a reduction or inhibition of hyaluronan synthesis in the retrobulbar cavity of the subject. In another embodiment, the treatment results in a reduction or inhibition of adipogenesis in the retrobulbar cavity of the subject. In another embodiment, the treatment results in a reduction in the levels of IL-6, IL-16, and / or RANTES in the serum of the subject.

[0022] In another specific embodiment, the IGF-1R-expressing cells are fibroblasts, B lymphocytes, and / or T lymphocytes.

[0023] In another embodiment, IGF-1R is overexpressed by the retrobulbar cells in the subject as compared to retrobulbar cells from a healthy subject or a subject not diagnosed with an ocular inflammatory condition. In another embodiment, the frequency of cells expressing IGF-1R in the subject is increased as compared to cells from a healthy subject or a subject not diagnosed with an ocular inflammatory condition. In another embodiment, the frequency of IGF-1R-expressing cells is measured by flow cytometry or immunohistochemistry.

[0024] In some embodiments, the treatment results in disease modification in the subject.

[0025] In some embodiments, the conjugate comprises wild-type insulin-like growth factor 1 (IGF-1) (SEQ ID NO: 3), wild-type insulin (SEQ ID NOs: 10 and 11), or wild-type insulin-like growth factor 2 (IGF-2) (SEQ ID NO: 12). In some embodiments, the conjugate comprises a variant of wild-type IGF-1 (SEQ ID NO: 3), a variant of wild-type insulin (SEQ ID NOs: 10 and 11), or a variant of wild-type IGF-2 (SEQ ID NO: 12). In some embodiments, the variant of wild-type IGF1 is at least 90% identical to IGF-1 (SEQ ID NO: 3), the variant of wild-type insulin is at least 90% identical to insulin (SEQ ID NOs: 10 and 11), or the variant of wild-type IGF-2 is at least 90% identical to IGF-2 (SEQ ID NO: 12).

[0026] In some embodiments, the conjugate comprises a variant of IGF-1 with a reduced binding affinity for an IGF-binding protein (IGFBP) compared to wild-type IGF-1 (SEQ ID NO: 3), or a variant of IGF-2 with a reduced binding affinity for an IGF-binding protein compared to wild-type IGF-2 (SEQ ID NO: 12). In some embodiments, the variant of IGF-1 has an increased affinity for IGF-1R compared to wild-type IGF-1 (SEQ ID NO: 3), or the variant of IGF-2 has an increased affinity for IGF-1R compared to wild-type IGF-2 (SEQ ID NO: 12). In certain embodiments, the conjugate comprises 765IGF (SEQ ID NO: 2), IGF132 (SEQ ID NO: 4), long-R3-IGF-1 (SEQ ID NO: 5), R3-IGF-1 (SEQ ID NO: 6), des(1-3)-IGF-1 (SEQ ID NO: 7), long-IGF-1 (SEQ ID NO: 8), or long-G3-IGF-1 (SEQ ID NO: 9).

[0027] In some embodiments, the IGF-1R ligand, or a portion or variant thereof, comprises a leader sequence. In certain embodiments, the leader sequence comprises SEQ ID NO: 1.

[0028] In certain embodiments, the conjugate comprises 765IGF (SEQ ID NO: 2).

[0029] In some embodiments, the disease modifier of the conjugate comprises a cytotoxic agent.

[0030] In some embodiments, the cytotoxic agent comprises a chemotherapeutic agent. In certain embodiments, the chemotherapeutic agent is amsacrine, azacitidine, bleomycin, busulfan, capecitabine, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, cyclophosphamide, cytarabine, dactinomycin, daunorubicin, dacarbazine, docetaxel, doxorubicin, epirubicin, estramustine, etoposide, floxuridine, fludarabine, fluorouracil, gemcitabine, hexamethylmelamine, idarubicin, ifosfamide, irinotecan, lomustine, mechlorethamine, melphalan, mercaptopurine, methotrexate, mitomycin C, mitotane, mitoxantrone, oxaliplatin, paclitaxel, pemetrexed, pentostatin, plicamycin, procarbazine, raltitrexed, semustine, streptozocin, temozolomide, teniposide, thioguanine, thiotepa, topotecan, trimethoprim, valrubicin, vincristine, vinblastine, vindesine, or vinorelbine. In another specific embodiment, the chemotherapeutic agent is methotrexate.

[0031] In some embodiments, the cytotoxic agent comprises a toxin. In certain embodiments, the toxin is Clostridium perfringens enterotoxin, diphtheria toxin, ricin A chain, deglycosylated ricin A chain, Pseudomonas exotoxin, A chain toxin, ribosome-inactivating protein, α-sarcin, aspergillin, abrin, restrictocin, bacterial endotoxin, the lipid A moiety of bacterial endotoxin, cholera toxin, or ribonuclease. In certain embodiments, the toxin comprises Clostridium perfringens enterotoxin, or a part or variant thereof. In certain embodiments, the conjugate comprises SEQ ID NO: 14, or SEQ ID NO: 15. In other specific embodiments, the toxin comprises diphtheria toxin, or a part or variant thereof. In certain embodiments, the conjugate comprises SEQ ID NO: 13, or SEQ ID NO: 16.

[0032] In other embodiments, the disease modifying agent of the conjugate of the invention is selected from glucocorticoids (dexamethasone), corticosteroids (methylprednisolone, prednisone, triamcinolone acetonide), TSHR inhibitors, mycophenolate mofetil, simvastatin, metformin, phenformin, cyclosporine, rapamycin or other mTOR inhibitors, or azathioprine.

[0033] In some embodiments, the subject having TED has Graves' hyperthyroidism and / or has at least 3, or at least 4 CAS.

[0034] In some embodiments, a subject having an ophthalmic inflammatory condition treated according to the methods described herein has not previously received treatment for the ophthalmic inflammatory condition, has previously received treatment for the ophthalmic inflammatory condition, has relapsed from a previous treatment for the ophthalmic inflammatory condition, or has been refractory to a previous treatment for the ophthalmic inflammatory condition.

[0035] In some embodiments, the previous treatment included administration of an IGF-1R inhibitor to the subject.

[0036] In certain embodiments, the IGF-1R inhibitor was an antibody. In another specific embodiment, the antibody was teprotumumab, ganitumab, figitumumab, MEDI-573, cicutumumab, daratumumab, robatumumab, AVE1642, BIIB022, xentuzumab, isothiazumab, lonigutumab, VB421, VRDN-001, VRDN-002, or VRDN-003.

[0037] In certain embodiments, the previous treatment included administration of an antibody, or antibody fragment, that targets the neonatal Fc receptor (FcRn). In certain embodiments, the antibody, or antibody fragment, is batoclimab, IMVT-1402, or efgartigimod.

[0038] In certain embodiments, previous treatment included administration of an antibody that targets IL-6. In certain embodiments, the antibody is TOUR006.

[0039] In certain embodiments, the IGF-1R inhibitor was a small molecule. In another certain embodiment, the small molecule was linsitinib, picropodophyllin, AXL1717, BMS-754807, BMS-536924, BMS-554417, GSK1838705A, GSK1904529A, NVP-AEW541, NVP-ADW742, GTx-134, AG1024, KW-2450, PL-2258, NVP-AEW541, NSM-18, AZD3463, AZD9362, BI885578, BI893923, TT-100, XL-228, or A-928605.

[0040] In some embodiments, the IGF-1R inhibitor was a radioimmunoconjugate. In certain embodiments, the radioimmunoconjugate was FPI-1434.

[0041] In some embodiments, the IGF-1R inhibitor was an antibody-drug conjugate. In certain embodiments, the antibody-drug conjugate was W0101.

[0042] In some embodiments, the conjugate is administered in combination with one or more other therapies. In certain embodiments, the one or more other therapies include glucocorticoids (dexamethasone), corticosteroids (methylprednisolone, prednisone, triamcinolone acetonide), rituximab or other anti-CD20 antibodies, tocilizumab, TOUR006, or other anti-IL-6 antibodies, selenium, infliximab, adalimumab, or other anti-tumor necrosis factor (TNF) α antibodies, thyroid stimulating hormone receptor (TSHR) inhibitors, orbital radiotherapy, vascular endothelial growth factor (VEGF) inhibitors, mycophenolate mofetil, isocablimab or other anti-CD40 antibodies, IMVT-1401, batoclimab, efgartigimod, or other neonatal crystallizable fragment (Fc) receptor (FcRN) inhibitors, simvastatin, metformin, phenformin, cyclosporine, azathioprine, methotrexate, rapamycin, or other mammalian targets of rapamycin (mTOR) inhibitors, and / or one or more of intravenous immunoglobulin therapy.

[0043] In some embodiments, the conjugate is administered at a dose of about 0.05, 0.10, 0.20, 0.40, 0.80, 1.0, 1.5, 1.6, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, or 10.0 microequivalents / kg body weight, or in a dose range of about 0.05 - 0.5, 0.5 - 1.0, 1.0 - 1.5, 1.5 - 2.0, 2.0 - 2.5, 2.5 - 3.0, 3.0 - 3.5, 3.5 - 4.0, 4.0 - 4.5, 4.5 - 5.0, 5.0 - 5.5, 5.5 - 6.0, 6.0 - 6.5, 6.5 - 7.0, 7.0 - 7.5, 7.5 - 8.0, 8.0 - 8.5, 8.5 - 9.0, 9.0 - 9.5, or 9.5 - 10.0 microequivalents / kg body weight.

[0044] In some embodiments, the conjugate is administered at a dose of about 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13.0, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14.0, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, 15.0, 15.1, 15.2, 15.3, 15.4, 15.5, 15.6, 15.7, 15.8, 15.9, 16.0, 16.1, 16.2, 16.3, 16.4, or 16.5 mg / kg body weight, or in the range of about 0.05 - 0.5, 0.5 - 1.0, 1.0 - 1.5, 1.5 - 2.0, 2.0 - 2.5, 2.5 - 3.0, 3.0 - 3.5, 3.5 - 4.0, 4.0 - 4.5, 4.5 - 5.0, 5.0 - 5.5, 5.5 - 6.0, 6.0 - 6.5, 6.5 - 7.0, 7.0 - 7.5, 7.5 - 8.0, 8.0 - 8.5, 8.5 - 9.0, 9.0 - 9.5, 9.5 - 10.0, 10.0 - 10.5, 10.It is administered in a dosage range of 5 to 11.0, 11.0 to 11.5, 11.5 to 12.0, 12.0 to 12.5, 12.5 to 13.0, 13.0 to 13.5, 13.5 to 14.0, 14.0 to 14.5, 14.5 to 15.0, 15.0 to 15.5, 15.5 to 16.0, or 16.0 to 16.5 mg / kg body weight.

[0045] In some embodiments, the conjugate is administered at a dosage that is the maximum tolerated dose (MTD).

[0046] In some embodiments, the conjugate is administered daily, every other day, once every 3 days, once every 4 days, once every 5 days, once every 6 days, once a week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once a month, once every 2 months, or once every 3 months.

[0047] In some embodiments, the conjugate is administered via intravenous, subcutaneous, intravitreal injection, via retrobulbar injection, or via an eye dropper.

[0048] In some embodiments, the methods of the invention do not cause unacceptable hyperglycemia in a subject.

[0049] In certain embodiments, provided herein is a method for treating TED in a subject in need thereof by administering an effective amount of a conjugate, said conjugate comprising 765IGF (SEQ ID NO: 2), and methotrexate.

[0050] These, and other embodiments, are described in full below.

DETAILED DESCRIPTION OF THE INVENTION

[0051] The subject matter described herein relates to a method of treating ophthalmic inflammatory conditions using a targeted therapy directed to IGF-1R, comprising an IGF-1R ligand, or a variant thereof, in combination with a disease modifying agent.

[0052] Here, the subject matter of the present disclosure will be described more fully. However, many modifications and other embodiments of the subject matter of the present disclosure described herein will come to mind to those skilled in the art regarding the benefits of the teachings presented in the foregoing description. Accordingly, it is to be understood that the subject matter of the present disclosure is not limited to the specific embodiments disclosed, and that modifications and other embodiments are intended to be included within the scope of the appended claims. In other words, the subject matter described herein encompasses all alternatives, modifications, and equivalents. If one or more of the incorporated documents, patents, and similar materials are different from or conflict with this application, including but not limited to defined terms, the use of terms, the described techniques, etc., this application controls. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. If one or more of the incorporated documents, patents, and similar materials are different from or conflict with this application, including but not limited to defined terms, the use of terms, the described techniques, etc., this application controls.

[0053] I. Definitions As used herein, a patient, or subject, etc. is any mammal suffering from an inflammatory condition of the eye. As used herein, the term "mammal" includes, but is not limited to, humans, mice, rats, guinea pigs, monkeys, dogs, cats, horses, cows, pigs, and sheep.

[0054] As used herein, the term "conjugate" refers to a molecule comprising an IGF-1R ligand, or a portion or variant thereof, and a disease modifying agent.

[0055] As used herein, the term "cytotoxic agent" refers to any agent that can prevent, delay, reduce, and / or reverse the activity, severity, and / or progression of a disease when treated according to the methods described herein. Any suitable cytotoxic agent that results in cell death can be used in conjugates and in methods of treating ophthalmic inflammatory conditions.

[0056] As used herein, the term "residue" or "residue of" a chemical moiety or compound refers to the chemical moiety or compound attached to a molecule, whereby at least one covalent bond through the attachment replaces at least one atom of the original chemical moiety or compound, resulting in a residue of the chemical moiety or compound within the molecule.

[0057] As used herein, a subject is "refractory" to a previous treatment if it fails to achieve a response to a therapy as determined to be therapeutically ineffective, e.g., if it fails to reach any clinical endpoint, including but not limited to response, prolongation of the duration of response, prolongation of the disease-free survival period, prolongation of the recurrence-free survival period, and prolongation of the progression-free survival period.

[0058] As used herein, "and / or" refers to one or more possible combinations of the associated listed items, as well as the absence of combinations when interpreted in the alternative (or "or"), and includes both.

[0059] As used herein, the term "about" when referring to a measurable value such as an amount of a compound or agent, of the present subject matter, dosage, time, temperature, etc., means including variations of ±20%, ±10%, ±5%, ±1%, ±0.5%, or ±0.1% of the specified amount.

[0060] As used herein, conditional language used herein, such as, among others, "can", "could", "might", "may", "for example", etc., generally conveys that a particular embodiment includes a particular feature, element, and / or step while other embodiments do not include the particular feature, element, and / or step, unless otherwise stated or otherwise understood within the context in which it is used. Thus, such conditional language is generally not intended to imply that a feature, element, and / or step is required in any way for one or more embodiments or that one or more embodiments necessarily include logic for determining whether these features, elements, and / or steps are included in or implemented in any particular embodiment, regardless of author input or indication. The terms "comprising", "including", "having", etc. are synonymous and are used in an inclusive, open-ended fashion and do not exclude additional elements, features, acts, operations, etc. Also, the term "or", when used, for example, to connect a list of elements, is used in its inclusive sense (and not in its exclusive sense) to mean one, some, or all of the elements in the list.

[0061] Definitions of additional terms may be set forth below.

[0062] II. Compositions Therapeutic agent This application provides a method of treating an ophthalmic inflammatory condition in a subject in need thereof, the method comprising administering to the subject an effective amount of a conjugate, the conjugate comprising an IGF-1R ligand, or a portion or variant thereof, and a disease modifying agent.

[0063] In this embodiment or any of the embodiments of the present invention, the conjugate can comprise a chemical conjugate in which an IGF-1R ligand and a disease modifying agent are chemically linked together either directly or via a chemical linker. In other embodiments, the conjugate is a recombinant in which the conjugate is expressed as a single polypeptide. When the conjugate is a recombinant conjugate, the translated conjugate preferably comprises a toxin, or a portion or variant thereof, linked to the IGF-1R ligand via a peptide bond.

[0064] In certain embodiments, the conjugate is the fusion protein described in U.S. Patent No. 9,675,671, which is hereby incorporated by reference in its entirety.

[0065] IGF-1R ligand IGF-1R is a heterotetramer consisting of two extracellular ligand-binding α subunits with kinase activity that mediates signal transduction and two transmembrane β subunits. The natural ligands for IGF-1R are IGF-1, IGF-2, and insulin. IGF-1R has the highest affinity for IGF-1, followed by an affinity for IGF-2, and can bind insulin with a 50 - 100-fold lower affinity. IGF-1R can also form hybrid receptors by dimerization with the insulin receptor. See Hakuno et al. J Mol Endocrinol. 61(1):T69-T86(2018).

[0066] In certain embodiments, the IGF-1R ligands in the conjugates of the invention include wild-type IGF-1 (SEQ ID NO: 3), wild-type insulin (SEQ ID NOs: 10 and 11), and mature insulin consists of two chains connected by a disulfide bond between chain A corresponding to SEQ ID NO: 10 and chain B corresponding to SEQ ID NO: 11, and thus consists of the listing of the two SEQ ID NOs, or wild-type IGF-2 (SEQ ID NO: 12). In other embodiments, the IGF-1R ligands in the conjugate include variants of wild-type IGF-1 (SEQ ID NO: 3), variants of wild-type insulin (SEQ ID NOs: 10 and 11), or variants of wild-type IGF-2 (SEQ ID NO: 12). In certain embodiments, the variant of wild-type IGF-1 is at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical to IGF-1 (SEQ ID NO: 3), the variant of said wild-type insulin is at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical to insulin (SEQ ID NOs: 10 and 11), or said variant of wild-type IGF-2 is at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical to IGF-2 (SEQ ID NO: 12).

[0067] In another specific embodiment, the IGF-1R ligand in the conjugate is a variant of IGF-1 having a reduced binding affinity for IGFBP compared to wild-type IGF-1 (SEQ ID NO: 3), or a variant of IGF-2 having a reduced binding affinity for IGFBP compared to wild-type IGF-2 (SEQ ID NO: 12). IGFBP belongs to a family of at least six proteins that bind IGF-1 and IGF-2 with high affinity. IGFBP binds to most of the circulating IGF, increases its half-life, regulates its bioavailability, and generally inhibits the ability to bind to IGF receptors. See Am J Physiol Endocrinol Metab., 278(6):E967-76 (2000), and Allard et al. Front Endocrinol (Lausanne). 9;9:117(2018). Thus, variants of IGF-1 or IGF-2 with reduced binding to IGFBP have higher biological activity in vivo.

[0068] IGF-1 variants with reduced binding affinity for IGFBP are known in the art and include IGF132 (U.S. Patent No. 4,876,242) (the first 17 amino acids of the B chain of insulin (SEQ ID NO: 11) replace the first 16 amino acids of human IGF-1 (SEQ ID NO: 3)), R3-IGF-1 (SEQ ID NO: 6) (glutamic acid at position 3 of native human IGF-1 (SEQ ID NO: 3) is replaced by arginine), and des(1-3)IGF-1 (SEQ ID NO: 7) (lacking the first three amino acids of human IGF-1 (SEQ ID NO: 3)). R3-IGF-1 and des(1-3)IGF-1 are described in Francis et al., J Mol Endocrinol. 8(3):213-23 (1992). In some embodiments, the conjugate comprises IGF132 (SEQ ID NO: 4), R3-IGF-1 (SEQ ID NO: 6), or des(1-3)-IGF-1 (SEQ ID NO: 7).

[0069] In another embodiment, a variant of IGF-1 has a higher affinity for IGF-1R than wild-type IGF-1 (SEQ ID NO: 3), or a variant of IGF-2 has a higher affinity for IGF-1R than wild-type IGF-2 (SEQ ID NO: 12).

[0070] In another embodiment, an IGF-1R ligand, or a portion or variant thereof, comprises a leader sequence. The leader sequence can incorporate a tag, such as a polyhistidine tag, to facilitate protein purification and provide a site for conjugation of a disease modifying agent. In certain embodiments, the leader sequence comprises SEQ ID NO: 1.

[0071] In some embodiments, the IGF-1R ligand in the conjugate comprises 765IGF (SEQ ID NO: 2), long-R3-IGF-1 (SEQ ID NO: 5), long-IGF-1 (SEQ ID NO: 8), or long-G3-IGF-1 (SEQ ID NO: 9). 765IGF, long-R3-IGF-1, long-IGF-1, and long-G3-IGF-1 have an N-terminal leader sequence that, as described above, facilitates protein purification and provides a site for conjugation of a disease modifying agent. 765IGF (SEQ ID NO: 2) comprises SEQ ID NO: 1, followed by R3-IGF-1 (SEQ ID NO: 6), long-R3-IGF-1 (SEQ ID NO: 5) comprises the first 11 amino acids of methionylbuta growth hormone, followed by a Val-Asn dipeptide, followed by R3-IGF-1 (SEQ ID NO: 6), long-IGF-1 (SEQ ID NO: 8) comprises the first 11 amino acids of methionylbuta growth hormone, followed by a Val-Asn dipeptide, followed by human IGF-1 (SEQ ID NO: 3), and long-G3-IGF-1 comprises the first 11 amino acids of methionylbuta growth hormone, followed by a Val-Asn dipeptide, followed by a variant of human IGF-1 in which the glutamic acid at position 3 of native human IGF-1 (SEQ ID NO: 3) is replaced with glycine. In some embodiments, the conjugate comprises 765IGF (SEQ ID NO: 2).

[0072] disease modifying agent As used herein, the term "disease modifying agent" refers to any agent that can prevent, delay, reduce, and / or reverse the activity, severity, and / or progression of a disease when treated according to the methods described in the present invention.

[0073] In certain embodiments, the disease modifying agent of the conjugate is a cytotoxic agent. Any suitable cytotoxic agent that causes cell death can be used in the conjugates of the present invention, and in methods of treating ophthalmic inflammatory conditions.

[0074] In certain embodiments, the cytotoxic agent is a chemotherapeutic agent. Any suitable chemotherapeutic agent that causes cell death can be used in the conjugates of the present invention, and in methods of treating ophthalmic inflammatory conditions. For example, in certain embodiments, the chemotherapeutic agent is amsacrine, azacitidine, bleomycin, busulfan, capecitabine, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, cyclophosphamide, cytarabine, dactinomycin, daunorubicin, dacarbazine, docetaxel, doxorubicin, epirubicin, estramustine, etoposide, floxuridine, fludarabine, fluorouracil, gemcitabine, hexamethylmelamine, idarubicin, ifosfamide, irinotecan, lomustine, mechlorethamine, melphalan, mercaptopurine, methotrexate, mitomycin C, mitotane, mitoxantrone, oxaliplatin, paclitaxel, pemetrexed, pentostatin, plicamycin, procarbazine, raltitrexed, semustine, streptozocin, temozolomide, teniposide, thioguanine, thiotepa, topotecan, trimetrexate, valrubicin, vincristine, vinblastine, vindesine, or vinorelbine. In certain embodiments, the chemotherapeutic agent is methotrexate.

[0075] In certain embodiments, the conjugate comprises two or more cytotoxic agents conjugated to an IGF-1R ligand. In certain aspects, the conjugate can comprise from 1 to 12 cytotoxic agents, or from 6 to 10 cytotoxic agents, or about 8 cytotoxic agents. In certain aspects, the conjugate can comprise from 1 to 12 covalently bound cytotoxic agents, or from 6 to 10 covalently bound cytotoxic agents, or about 8 covalently bound cytotoxic agents. In certain aspects, the cytotoxic agent is covalently bound to any available position on the IGF-1R ligand. In certain aspects, the cytotoxic agent is covalently bound to any available lysine residue. In certain aspects, the cytotoxic agent, if present, is covalently bound to any available lysine in the leader sequence.

[0076] In other embodiments, the cytotoxic agent is a toxin. Any suitable toxin that causes cell death can be used in the conjugates of the invention and in methods of treating ophthalmic inflammatory conditions. The toxin can be derived from a plant, fungus, or bacterium. For example, in certain embodiments, the toxin is Clostridium perfringens enterotoxin, diphtheria toxin, ricin A chain, deglycosylated ricin A chain, Pseudomonas exotoxin, A chain toxin, ribosome-inactivating protein, alpha-sarcin, aspergillin, abrin, restrictocin, bacterial endotoxin, the lipid A portion of bacterial endotoxin, cholera toxin, or ribonuclease. In certain embodiments, the toxin comprises Clostridium perfringens enterotoxin, or a portion of a variant thereof. In certain embodiments, the conjugate comprises SEQ ID NO: 14, or SEQ ID NO: 15. In other certain embodiments, the toxin comprises diphtheria toxin, or a portion or variant thereof. In certain embodiments, the conjugate comprises SEQ ID NO: 13, or SEQ ID NO: 16.

[0077] In another specific embodiment, the disease-modifying agent of the conjugate of the present invention is selected from glucocorticoids (dexamethasone), corticosteroids (methylprednisolone, prednisone, triamcinolone acetonide), TSHR inhibitors, mycophenolate mofetil, simvastatin, metformin, phenformin, cyclosporine, rapamycin or other mTOR inhibitors, or azathioprine.

[0078] Table 1 provides a list of specific sequences referred to herein.

Table 1-1

Table 1-2

[0079] Pharmaceutical composition The conjugate of the present invention can be formulated in a pharmaceutical composition for use by the methods described herein. In some embodiments, the pharmaceutical composition comprises an effective amount of the conjugate of the present invention and a pharmaceutically acceptable carrier or vehicle. Such pharmaceutical compositions can be formulated to be suitable for administration to a subject and can be in any form that enables the composition to be administered to the subject.

[0080] The materials used in the preparation of the pharmaceutical composition can be non-toxic in the amounts used. It will be apparent to those skilled in the art that the optimal dosage of the active ingredient in the pharmaceutical composition depends on various factors. Relevant factors include, but are not limited to, the type of subject (e.g., human), the overall health of the subject, the type of condition the subject has, the use of the composition as part of a multi-drug regimen, the specific form of the composition of the present invention, and the method of administration. The pharmaceutical composition comprises an effective amount of the composition of the present invention such that a suitable dosage is obtained.

[0081] The term "carrier" refers to a diluent, adjuvant, or excipient in which the composition of the present invention is administered. Any auxiliary agents, stabilizers, thickeners, lubricants, and coloring agents can be used. In one embodiment, when administered to a subject, the composition of the present invention, and the pharmaceutically acceptable carrier, are sterilized. Water may be a carrier when the composition of the present invention is administered intravenously. Physiological saline, as well as aqueous solutions of dextrose and glycerol, can also be used as liquid carriers, particularly for injection solutions. The composition can also contain a small amount of a pH buffer, if desired.

[0082] Any liquid composition, solution, suspension, or other similar form of the present invention can also contain one or more of the following: sterile diluents such as water for injection, physiological saline, physiological saline solution, Ringer's solution, isotonic sodium chloride, solvents, or suspension media that can function as such, fixed oils such as synthetic mono- or diglycerides, polyethylene glycol, glycerol, cyclodextrin, propylene glycol, or other solvents, antibacterial agents such as benzyl alcohol or methylparaben, antioxidants such as ascorbic acid or sodium sulfite, chelating agents such as ethylenediaminetetraacetic acid, buffers such as acetate, citrate, or phosphate, agents for adjusting the pH such as hydrochloric acid, and agents for adjusting the tonicity such as sodium chloride or dextrose. The parenteral composition can be enclosed in ampoules, disposable syringes, or multiple-dose vials made of glass, plastic, or other materials. In some embodiments, physiological saline is an adjuvant. The injectable composition may be sterilized.

[0083] The composition can take the form of a solution, suspension, tablet, pill, pellet, capsule, capsule containing a liquid, powder, sustained-release formulation, suppository, emulsion, aerosol, spray, suspension, or any other form suitable for use. Examples of suitable pharmaceutical carriers are described in Remington’s Pharmaceutical Sciences by E.W. Martin.

[0084] In some embodiments, the compositions of the invention are formulated according to routine procedures as pharmaceutical compositions adapted for intravenous administration to a human subject. Typically, the carrier, or vehicle, for intravenous administration is a sterile isotonic aqueous buffer solution. Optionally, the composition may also contain solubilizing agents. The composition for intravenous administration may optionally contain a local anesthetic, such as lignocaine, to reduce pain at the injection site. Generally, the components are supplied in unit dosage forms, separately or mixed together, as a dry lyophilized powder, or an anhydrous concentrate, in a sealed container, such as an ampoule, or sachet, indicating the amount of the active agent, for example. When the composition of the invention is administered by infusion, it can be dispensed using an infusion bottle containing, for example, sterile pharmaceutical grade water, or saline. When the composition of the invention is administered by injection, an ampoule of sterile water for injection, or saline, may be provided so that the components can be mixed before administration.

[0085] The pharmaceutical compositions can be prepared using methodologies well known in the pharmaceutical art. For example, a composition intended to be administered by injection can be prepared by combining the composition of the invention with water to form a solution. A surfactant can be added to facilitate the formation of a homogeneous solution, or suspension. The surfactant is a conjugate that can interact non-covalently with the composition of the invention to facilitate the dissolution, or homogeneous suspension, of the composition of the invention in an aqueous delivery system.

[0086] III. Methods Method for preparing the IGF-1R conjugate Methods for generating the conjugates of the invention are known in the art.

[0087] The nucleotide sequence encoding the IGF-1R ligand is generated by standard recombinant DNA techniques or by protein synthesis techniques, cloned into an appropriate expression vector using standard molecular biology techniques, expressed in bacterial cells, insect cells, or mammalian cells, and can be purified by any method known in the art for protein purification.

[0088] The conjugates of the present invention comprising an IGF-1R ligand and a chemotherapeutic agent can be made by standard chemistry and protein conjugation techniques and are described in U.S. Patent No. 7,811,982, U.S. Patent No. 9,675,671, and U.S. Patent No. 9,801,923.

[0089] The conjugates of the present invention comprising an IGF-1R ligand and a toxin can be made as a fusion protein by standard recombinant DNA techniques and are described in U.S. Patent No. 8,017,102.

[0090] Methods of Treatment Provided herein is a method of treating an inflammatory condition of the eye using a conjugate comprising an IGF-1R ligand, or a portion or variant thereof, and a disease modifier.

[0091] In some embodiments, the inflammatory condition of the eye is TED, uveitis, scleritis, keratitis, or conjunctivitis.

[0092] In some embodiments, the inflammatory condition of the eye is an orbital inflammatory disease. In certain embodiments, the orbital inflammatory disease is idiopathic orbital inflammation, orbital inflammatory pseudotumor, orbital myositis, inflammatory orbital cellulitis, perineuritis of the optic nerve, episcleritis, diffuse orbital inflammation, orbital apex syndrome, or sclerosing orbital inflammation.

[0093] In certain embodiments, the inflammatory condition of the eye is TED. TED may also be referred to as Graves ophthalmopathy, thyroid-associated ophthalmopathy, Graves eye disease, Graves orbitopathy, or thyroid inflammatory eye disease.

[0094] In certain embodiments, the subject matter described herein is directed to a method for treating an ophthalmic inflammatory condition in a subject, the method comprising administering to the subject a conjugate comprising an IGF-1R ligand, or a portion or variant thereof, and a cytotoxic agent, wherein the IGF-1R ligand, or a portion or variant thereof, comprises SEQ ID NO:2, the cytotoxic agent is methotrexate, methotrexate is covalently bound to a lysine of SEQ ID NO:2, and the ophthalmic inflammatory condition is selected from the group consisting of Graves' ophthalmopathy, uveitis, scleritis, keratitis, conjunctivitis, and orbital inflammatory disease. In certain aspects of these embodiments, the conjugate is LX-101 (the conjugate described above, wherein the IGF-1R ligand is SEQ ID NO:2, the cytotoxic agent is methotrexate, and at least 6 to a maximum of 10, or at least 6 to a maximum of 9, or at least 7 to a maximum of 9, or at least 8 to a maximum of 9 methotrexate residues are each covalently bound to a lysine residue of SEQ ID NO:2).

[0095] In certain embodiments, the number of methotrexate residues per conjugate is 6, 7, 8, 9, or 10. In certain embodiments, the average number of methotrexate residues per conjugate in the composition is 6, 7, 8, 9, or 10.

[0096] Methods for measuring clinical efficacy As used herein, the terms "subject" and "patient" are used interchangeably.

[0097] In some embodiments, a method for treating a subject's TED includes administering the conjugate of the invention to a subject in need thereof, resulting in a reduction in proptosis in the subject. Proptosis, also known as exophthalmos, is the protrusion of the eyeball, which causes bulging of the eyes and can affect one or both eyes. In TED, proptosis is caused by the expansion of orbital fat and the extraocular muscles. See Khong et al., Br J Ophthalmol, 100(1):142-50 (2016). In some embodiments, the reduction in proptosis is greater than 2 mm, for example, 2.2 mm, 2.4 mm, 2.5 mm, 2.6 mm, 2.8 mm, 3 mm, 3.2 mm, 3.4 mm, 3.5 mm, 3.6 mm, 3.8 mm, 4 mm, 4.1 mm, 4.2 mm, 4.3 mm, 4.4 mm, 4.5 mm, 4.6 mm, 4.7 mm, 4.8 mm, 4.9 mm, 5 mm, or more than 5 mm. In certain embodiments, the proptosis is reduced by at least 2 mm, at least 3 mm, or at least 4 mm. In another embodiment, the reduction in proptosis is related to a reduction in the volume of the extraocular muscles and / or the volume of orbital fat in the subject. In another embodiment, the reduction in proptosis is evaluated by a proptometer or orbital imaging. In certain embodiments, the orbital imaging is computed tomography scan or magnetic resonance imaging.

[0098] In some embodiments, a method for treating TED in a subject comprises administering a conjugate of the invention to a subject in need thereof, resulting in a reduction of CAS in the subject. CAS consists of seven components (orbital pain or discomfort, pain upon upward or downward gaze, eyelid erythema, eyelid swelling, conjunctival injection, conjunctival edema, caruncle erythema / swelling), each of which is scored as 1 (present) or 0 (absent). CAS is the sum of all present items and ranges from 0 to 7, a score of 0 or 1 constitutes an inactive disease, a score of 3 or more constitutes an active disease, and a score of 7 constitutes a severe active disease. See Bartalena et al. Eur J Endocrinol. 185(4):G43-G67(2021). A change of two or more points is considered clinically significant. In some embodiments, the reduction of CAS is two or more points, for example, 3, 4, 5, 6, or 7 points. In one embodiment, the reduction of CAS is two or more points. In another embodiment, this is three or more points. In yet another embodiment, the reduction of CAS is four or more points. In certain embodiments, CAS is reduced by at least two points, or at least three points. In another specific embodiment, CAS is reduced to 1 or reduced to 0.

[0099] In some embodiments, a method for treating a subject's TED comprises administering a conjugate of the invention to a subject in need thereof, resulting in a reduction in the severity of diplopia in the subject. In another embodiment, the reduction in diplopia is measured by the Gorman subjective diplopia score. Diplopia is the medical term for double vision, which is evaluated by the Gorman subjective diplopia score and includes four categories: no diplopia (none, score 0), diplopia at the primary position of gaze when the patient is fatigued or awake (intermittent, score 1), diplopia at the extreme of gaze (unstable, score 2), and continuous diplopia at the primary position or reading position (constant, score 3). See Bartalena et al. Eur J Endocrinol, 158(3):273-85 (2008). An improvement of one grade or more is considered clinically significant. In certain embodiments, the diplopia is constant diplopia, intermittent diplopia, or indeterminate diplopia. In certain embodiments, the treatment results in the complete disappearance of diplopia. In another specific embodiment, the severity of diplopia in the subject is reduced by at least one grade.

[0100] In some embodiments, a method for treating TED in a subject comprises administering a conjugate of the invention to a subject in need thereof, resulting in an improvement in the subject's quality of life. The quality of life is measured by the GO-QoL assessment scale. The GO-QoL assessment scale is a self-administered questionnaire consisting of two subscales, each with eight questions. The visual function subscale assesses the impact of visual function on daily activities, and the appearance subscale assesses the impact on self-perceived appearance. Each question is scored from 0 to 2, and the total raw score is converted to a scale of 0 to 100, where 0 has the most adverse impact on quality of life and 100 has no adverse impact on quality of life. The raw scores from both subscales are also combined and converted to a 0 to 100 scale. See Terwee, et al. Br J Ophthalmol, 82(7):773-9 (1998). A change of eight points or more has been shown to be clinically significant. In certain embodiments, the improvement in quality of life is measured by the GO-QoL assessment scale, or the visual function subscale of GO-QoL, or the appearance subscale. In another particular embodiment, the quality of life is improved by at least eight points on the GO-QoL assessment scale, the visual function subscale of GO-QoL, or the appearance subscale of GO-QoL.

[0101] Optic neuropathy is a serious, vision-threatening complication of TED that can result from compression or stretching of the optic nerve. In some embodiments, a method for treating TED in a subject comprises administering a conjugate of the invention to a subject in need thereof, resulting in an improvement in optic neuropathy.

[0102] In some embodiments, a method for treating TED in a subject comprises administering a conjugate of the invention to a subject in need thereof, resulting in a reduction in retro-orbital edema.

[0103] Restriction of eye movement resulting from swelling of the muscles that control eye movement is another complication of TED. In some embodiments, a method for treating TED in a subject, including administering a conjugate of the invention to a subject in need thereof, results in an improvement in monocular adduction. In certain embodiments, the improvement in monocular adduction is measured by a light reflex test. In another specific embodiment, the improvement in monocular adduction is at least 10 degrees.

[0104] In some embodiments, a method for treating an inflammatory condition of the eye in a subject includes administering a conjugate of the invention to a subject in need thereof, resulting in a change or impairment in the function of IGF-1R-expressing cells in the subject. As used herein, a change or impairment refers to any change to the endogenous function of the IGF-1R-expressing cells in the inflammatory condition of the eye.

[0105] In some embodiments, a method for treating an inflammatory condition of the eye in a subject includes administering a conjugate of the invention to a subject in need thereof, resulting in a reduction in the number of IGF-1R-expressing cells in the subject. In certain embodiments, the reduction is caused by killing of the IGF-1R-expressing cells. Without being bound by theory, simple inhibition of the IGF-1R pathway may be insufficient as a completely curative treatment due to other, potentially redundant signaling, either by the disease effector continuing to act or being upregulated and resulting in compensatory pathways that can lead to adaptive resistance. Thus, targeted delivery of a disease-modifying agent, including an agent that results in killing of IGF-1R-expressing cells, may provide benefits beyond simple IGF-1R pathway inhibition (including being preferential) to bypass the effects of such potentially redundant and / or compensatory pathways.

[0106] In another specific embodiment, the IGF-1R-expressing cells are orbital fibroblasts (OF). OF are central to the pathophysiology of TED. Activated OF proliferate and differentiate into adipocytes and myofibroblasts, secrete extracellular matrix proteins such as hyaluronan, and cause soft tissue expansion, all of which contribute to the various clinical symptoms of TED. Hyaluronan is a salt of hyaluronic acid (HA). Hyaluronan and hyaluronic acid may be used interchangeably herein. Cytokines and inflammatory mediators produced by immune cells (e.g., T cells, B cells, mast cells, and monocytes) infiltrating the orbit can promote the differentiation of OF into myofibroblasts and adipocytes, leading to scarring, tissue remodeling, and tissue expansion. Myofibroblasts are contractile cells that express smooth muscle actin (αSMA). In some embodiments, the treatment results in a reduction or inhibition of hyaluronan synthesis in the retrobulbar space of the subject. In some embodiments, the treatment results in a reduction in the level of hyaluronan or hyaluronic acid. In other embodiments, the treatment results in a reduction or inhibition of adipogenesis in the retrobulbar space of the subject. In some embodiments, the treatment results in a reduction in the levels of IL-6, IL-16, and / or RANTES in the serum of the subject. In some embodiments, the treatment results in a reduction or inhibition of the differentiation of OF into myofibroblasts.

[0107] In another specific embodiment, the IGF-1R-expressing cells are fibroblasts, B lymphocytes, and / or T lymphocytes. Fibroblasts are bone marrow-derived fibroblast-like precursors that circulate peripherally and have both tissue remodeling and immunomodulatory properties. Without being bound by theory, it is thought that fibroblasts infiltrate the orbit, differentiate into OF, and may contribute to the pathophysiology of TED and potentially other inflammatory conditions of the eye. B lymphocytes produce pathogenic autoantibodies that can cause tissue damage and / or dysregulation of cell signaling. T lymphocytes activate autoreactive B cells and OF and can contribute to inflammation and tissue damage.

[0108] In some embodiments, IGF-1R is overexpressed by cells in the retrobulbar cavity compared to cells from healthy subjects or subjects not diagnosed with an ocular inflammatory condition. In some embodiments, the frequency of cells expressing IGF-1R in a subject is increased compared to cells from healthy subjects or subjects not diagnosed with an ocular inflammatory condition. In some embodiments, the frequency of IGF-1R-expressing cells is measured by flow cytometry or immunohistochemistry using techniques known in the art and cell surface markers.

[0109] In some embodiments, a method of treating an ocular inflammatory condition in a subject in need thereof using the conjugate of the invention results in disease modification. As used herein, disease modification can refer to any prevention, delay, reduction, and / or reversal of the activity, severity, and / or progression of a disease in a subject when treated according to the methods described in the present invention. Disease modification can result from a direct or indirect effect on the pathophysiology underlying the disease as a result of treatment with the conjugate of the invention.

[0110] Combination therapy In some embodiments, the conjugate of the invention is administered in combination with one or more other therapies. Any therapy used for the treatment of TED can be used in combination with the conjugate of the invention, including: glucocorticoids (dexamethasone), corticosteroids (methylprednisolone, prednisone, triamcinolone acetonide), rituximab or other anti-CD20 antibodies, tocilizumab, TOUR006, or other anti-IL-6 antibodies, selenium, infliximab, adalimumab, or other anti-TNFα antibodies, TSHR inhibitors, orbital radiotherapy, vascular endothelial growth factor (VEGF) inhibitors, mycophenolate mofetil, isocablimab or other anti-CD40 antibodies, IMVT-1401, batoclimab, efgartigimod or other FcRN inhibitors, simvastatin, metformin, phenformin, cyclosporine, azathioprine, methotrexate, rapamycin or mTOR inhibitors, and / or intravenous immunoglobulin therapy.

[0111] In some embodiments, the combination of agents disclosed herein allows one or more agents to be administered at a dosage level lower than the dosage at which the agent would have an effect when administered as a single agent.

[0112] Administration and Dosage The agents of the present invention may be administered at any clinically relevant dosage. Clinically relevant means that the dosage of the agent has an effect on the subject. In some embodiments, the conjugate of the present invention is administered at a dosage of about 0.05, 0.10, 0.20, 0.40, 0.80, 1.0, 1.5, 1.6, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, or 10.0 μequivalent / kg body weight, where μequivalent is equivalent to μmol of the chemotherapeutic agent moiety conjugated to the IGF-1R ligand.

[0113] In some embodiments, the conjugate of the present invention is administered in a dosage range of about 0.05 - 0.5, 0.5 - 1.0, 1.0 - 1.5, 1.5 - 2.0, 2.0 - 2.5, 2.5 - 3.0, 3.0 - 3.5, 3.5 - 4.0, 4.0 - 4.5, 4.5 - 5.0, 5.0 - 5.5, 5.5 - 6.0, 6.0 - 6.5, 6.5 - 7.0, 7.0 - 7.5, 7.5 - 8.0, 8.0 - 8.5, 8.5 - 9.0, 9.0 - 9.5, or 9.5 - 10.0 μequivalent / kg body weight, where μequivalent is equivalent to μmol of the chemotherapeutic agent moiety conjugated to the IGF-1R ligand.

[0114] In some embodiments, the conjugate of the invention is administered at a dose of about 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13.0, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14.0, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, 15.0, 15.1, 15.2, 15.3, 15.4, 15.5, 15.6, 15.7, 15.8, 15.9, 16.0, 16.1, 16.2, 16.3, 16.4, or 16.5 mg / kg body weight, where mg refers to the amount of IGF-1R ligand present in the conjugate.

[0115] In some embodiments, the conjugate of the present invention is administered in a dosage range of about 0.05 - 0.5, 0.5 - 1.0, 1.0 - 1.5, 1.5 - 2.0, 2.0 - 2.5, 2.5 - 3.0, 3.0 - 3.5, 3.5 - 4.0, 4.0 - 4.5, 4.5 - 5.0, 5.0 - 5.5, 5.5 - 6.0, 6.0 - 6.5, 6.5 - 7.0, 7.0 - 7.5, 7.5 - 8.0, 8.0 - 8.5, 8.5 - 9.0, 9.0 - 9.5, 9.5 - 10.0, 10.0 - 10.5, 10.5 - 11.0, 11.0 - 11.5, 11.5 - 12.0, 12.0 - 12.5, 12.5 - 13.0, 13.0 - 13.5, 13.5 - 14.0, 14.0 - 14.5, 14.5 - 15.0, 15.0 - 15.5, 15.5 - 16.0, or 16.0 - 16.5 mg / kg body weight, where mg refers to the amount of IGF-1R ligand present in the conjugate.

[0116] In some embodiments, the conjugate of the present invention is administered at the maximum tolerated dose (MTD). As used herein, MTD refers to the maximum dose of a drug that an individual patient can tolerate. In other words, the side effects in a given patient can determine the MTD. Side effects can limit the ability to administer a higher dose of treatment than the maximum tolerated dose. Thus, the MTD for a given patient may be lower than that indicated in the prescription information for treatment or that commonly used in clinical practice. The MTD may have limited or no clinical efficacy in a patient.

[0117] In some embodiments, the conjugate of the present invention is administered daily, every other day, once every 3 days, once every 4 days, once every 5 days, once every 6 days, once a week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once a month, once every 2 months, or once every 3 months.

[0118] The conjugates of the present invention can be administered by any convenient route, for example, by injection or bolus injection, by absorption through the epithelium or mucosal lining (e.g., oral mucosa, rectum, and intestinal mucosa, etc.). Administration can be systemic or local. For example, various delivery systems such as microparticles, microcapsules, capsules, etc. are known and can be useful for the administration of the compositions of the present invention. Administration methods include, but are not limited to, oral administration and parenteral administration, parenteral administration, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, sublingual, intranasal, intracerebral, intraventricular, subarachnoid, intravaginal, transdermal, rectal, inhalation, or topical to the ear, nose, eyes, or skin. The mode of administration is left to the discretion of the physician and depends in part on the site of the medical condition.

[0119] In some embodiments, the compositions of the present invention are administered parenterally. In some embodiments, the compositions of the present invention are administered intravenously. In another embodiment, the compositions of the present invention are administered by continuous infusion. In certain embodiments, the compositions of the present invention are administered by an infusion that lasts for 15 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, or 2 hours.

[0120] In some embodiments, it may be desirable to locally administer one or more of the compositions of the present invention to the area in need of treatment. This can be achieved, for example, but not limited to, local injection during surgery, for example, by local application in combination with a wound dressing after surgery, by injection, by catheter, by suppository, or by implant, and the implant is a porous, non-porous, or gelatinous material including a membrane such as a cyalastic membrane or fiber. In certain embodiments, one or more of the compositions of the present invention can be administered intraperitoneally.

[0121] In yet another embodiment, the compositions of the present invention can be delivered by a controlled release system.

[0122] In some embodiments, a pump can be used to deliver the compositions of the invention (see, e.g., Sefton, CRC Crit. Ref. Biomed. Eng. 1987, 14, 201; Buchwald et al., Surgery 1980, 88: 507; Saudek et al., N. Engl. J. Med. 1989, 321: 574). In some embodiments, the pump may be, but is not limited to, an insulin-like pump.

[0123] In some embodiments, the conjugates of the invention are administered intravenously, subcutaneously, intravitreally, via retrobulbar injection, or via an eye dropper.

[0124] In some embodiments, the methods of the invention do not cause unacceptable hyperglycemia in a subject. Hyperglycemia is another term for high blood sugar and can occur when there is insufficient insulin in the body or when the body cannot utilize insulin properly. Unacceptable hyperglycemia refers to grade 3 or higher adverse effects as determined by a treating physician and / or adverse effects that cannot be controlled with diabetes medications and that lead to discontinuation of treatment with the conjugates of the invention.

[0125] Patient population TED is a progressive condition having two distinct phases. The first phase is an acute, or active, inflammatory phase characterized by soft tissue expansion and lasting from 6 months to 3 years. The second phase is a chronic, or inactive, phase characterized by fibrosis and reduced inflammation. In some embodiments, the conjugates of the invention are administered to subjects diagnosed with active TED, acute TED, inactive TED, or chronic TED.

[0126] In some embodiments, the conjugates of the invention are administered to subjects diagnosed with moderate to severe TED, or visually threatening (very severe) TED. Moderate to severe TED includes patients with no visually threatening disease but whose eye disease has a sufficient impact on daily life to justify immunosuppression (in the case of active disease) or surgical intervention (in the case of active disease). These patients typically have two or more of the following: eyelid retraction ≥2 mm, moderate or severe soft tissue involvement, proptosis ≥3 mm above normal for race and gender, and intermittent or constant diplopia. Visually threatening TED includes patients with thyroid ophthalmopathy and / or corneal detachment. See Bartalena et al. Eur J Endocrinol. 185(4):G43-G67 (2021).

[0127] In some embodiments, the subject having TED has Graves' hyperthyroidism and / or has at least 3, or at least 4, CASs.

[0128] In some embodiments, a subject having an ophthalmic inflammatory condition treated according to the methods described herein has not previously received treatment for the ophthalmic inflammatory condition. In some embodiments, a subject having an ophthalmic inflammatory condition treated according to the methods described herein has previously received treatment for the ophthalmic inflammatory condition.

[0129] In some embodiments, the previous treatment included administration of an IGF-1R inhibitor to the subject.

[0130] In some embodiments, the IGF-1R inhibitor was an antibody. In certain embodiments, the antibody was teprotumumab, ganitumab, figitumumab, MEDI-573, siltuximab, daratumumab, robatumumab, AVE1642, BIIB022, xencizumab, isatuximab, lonigatumumab, VB421, VRDN-001, VRDN-002, or VRDN-003.

[0131] In some embodiments, previous treatment included administration of an antibody, or antibody fragment, that targets the neonatal Fc receptor (FcRn). In certain embodiments, the antibody is batoclimab, IMVT-1402, or efgartigimod.

[0132] In some embodiments, previous treatment included administration of an antibody that targets IL-6. In certain embodiments, the antibody is TOUR006.

[0133] In some embodiments, the IGF-1R inhibitor was a small molecule. In certain embodiments, the small molecule was linsitinib, picropodophyllin, AXL1717, BMS-754807, BMS-536924, BMS-554417, GSK1838705A, GSK1904529A, NVP-AEW541, NVP-ADW742, GTx-134, AG1024, KW-2450, PL-2258, NVP-AEW541, NSM-18, AZD3463, AZD9362, BI885578, BI893923, TT-100, XL-228, or A-928605.

[0134] In some embodiments, the IGF-1R inhibitor was a radioimmunoconjugate. In certain embodiments, the radioimmunoconjugate was FPI-1434.

[0135] In some embodiments, the IGF-1R inhibitor was an antibody-drug conjugate. In certain embodiments, the antibody-drug conjugate was W0101.

[0136] In some embodiments, the subject is experiencing a recurrence from a previous treatment for an ophthalmic inflammatory condition.

[0137] In some embodiments, the subject was refractory to previous treatment for an inflammatory condition of the eye. As used herein, a subject is "refractory" to previous treatment if the subject was unable to achieve a response to a therapy such that the therapy was determined to be therapeutically ineffective, e.g., unable to reach a clinical endpoint including any of response, prolongation of the duration of response, prolongation of disease-free survival, prolongation of recurrence-free survival, and prolongation of progression-free survival.

[0138] The subject matter described herein includes, but is not limited to, the following embodiments.

[0139] 1. A method of treating an inflammatory condition of the eye in a subject, the method comprising administering to the subject an effective amount of a conjugate, the conjugate comprising (i) an insulin-like growth factor 1 receptor (IGF-1R) ligand, or a portion or variant thereof, and (ii) a disease modifying agent.

[0140] 2. The method of embodiment 1, wherein the inflammatory condition of the eye is thyroid eye disease (TED), uveitis, scleritis, keratitis, conjunctivitis, or an orbital inflammatory disease.

[0141] 3. The method of embodiment 2, wherein the orbital inflammatory disease is idiopathic orbital inflammation, orbital inflammatory pseudotumor, orbital myositis, inflammatory orbital cellulitis, optic nerve sheath meningitis, episcleritis, diffuse orbital inflammation, orbital apex syndrome, or sclerosing orbital inflammation.

[0142] 4. Embodiment 2, wherein the inflammatory condition of the eye is TED.

[0143] 5. The method of embodiment 4, wherein the TED is active TED, acute TED, inactive TED, chronic TED, moderate to severe TED, or vision-threatening (very severe) TED.

[0144] 6. The method of embodiment 4 or 5, wherein the treatment results in a reduction in proptosis in the subject.

[0145] 7. The method according to embodiment 6, wherein the proptosis is reduced by at least 2 mm, at least 3 mm, or at least 4 mm.

[0146] 8. The method according to embodiment 6 or 7, wherein the reduction of the proptosis is evaluated by a proptometer or orbital imaging.

[0147] 9. The method according to embodiment 8, wherein the orbital imaging is a computed tomography (CT) scan or magnetic resonance imaging.

[0148] 10. The method according to any one of embodiments 6 to 9, wherein the reduction of the proptosis is related to a reduction in the volume of the extraocular muscles and / or the volume of the orbital fat.

[0149] 11. The method according to any one of embodiments 4 to 10, wherein the treatment results in a reduction in the clinical activity score (CAS) in the subject.

[0150] 12. The method according to embodiment 11, wherein the CAS is reduced by at least 2 points or at least 3 points.

[0151] 13. The method according to embodiment 11, wherein the CAS is reduced to 1 or reduced to 0.

[0152] 14. The method according to any one of embodiments 4 to 13, wherein the treatment results in a reduction in the severity of diplopia in the subject.

[0153] 15. The method according to embodiment 14, wherein the reduction in the severity of diplopia is measured by the Gorman subjective diplopia score.

[0154] 16. The method according to embodiment 14 or 15, wherein the diplopia is constant diplopia, intermittent diplopia, or variable diplopia.

[0155] 17. The method according to any one of embodiments 14 to 16, wherein the treatment results in a complete remission of diplopia.

[0156] 18. The method according to any one of embodiments 15 to 17, wherein the severity of diplopia in the subject is reduced by at least 1 grade.

[0157] 19. The method according to any one of embodiments 4 to 18, wherein the treatment results in an improvement in the quality of life of the subject.

[0158] 20. The method according to embodiment 19, wherein the improvement in the quality of life is measured by the Graves' ophthalmopathy quality of life (GO-QoL) assessment scale, or the visual function subscale of GO-QoL, or the appearance subscale of GO-QoL.

[0159] 21. The method according to embodiment 20, wherein the quality of life is improved by at least 8 points on the GO-QoL assessment scale, or the visual function subscale of GO-QoL, or the appearance subscale of GO-QoL.

[0160] 22. The method according to any one of embodiments 4 to 21, wherein the treatment results in an improvement in neuropathy.

[0161] 23. The method according to any one of embodiments 4 to 22, wherein the treatment results in a reduction in retro-orbital edema.

[0162] 24. The method according to any one of embodiments 4 to 23, wherein the treatment results in an improvement in monocular retraction movement.

[0163] 25. The method according to embodiment 24, wherein the improvement in monocular retraction movement is measured by a light reflex test.

[0164] 26. The method according to embodiment 25, wherein the improvement is at least 10 degrees.

[0165] 27. The method according to any one of embodiments 1 to 26, wherein the treatment results in a change or inhibition of the function of IGF-1R-expressing cells in the subject.

[0166] 28. The method according to any one of embodiments 1 to 26, wherein the treatment results in a decrease in the number of IGF-1R-expressing cells in the subject.

[0167] 29. The method according to embodiment 28, wherein the decrease is caused by killing the IGF-1R-expressing cells.

[0168] 30. The method according to any one of embodiments 27 to 29, wherein the IGF-1R-expressing cells are orbital fibroblasts (OF).

[0169] 31. The method according to any one of embodiments 4 to 30, wherein the treatment results in a reduction or inhibition of hyaluronan synthesis in the retrobulbar space.

[0170] 32. The method according to any one of embodiments 4 to 31, wherein the treatment results in a reduction or inhibition of adipogenesis in the retrobulbar space of the subject.

[0171] 33. The method according to any one of embodiments 4 to 32, wherein the treatment results in a reduction in the levels of interleukin (IL)-6, IL-16, and / or RANTES in the serum of the subject.

[0172] 34. The method according to any one of embodiments 27 to 29, wherein the IGF-1R-expressing cells are fibroblasts, B lymphocytes, and / or T lymphocytes.

[0173] 35. The method according to any one of embodiments 1 to 34, wherein the IGF-1R is overexpressed by retrobulbar cells in the subject compared to retrobulbar cells from a healthy subject or a subject not diagnosed with an ocular inflammatory condition.

[0174] 36. The method according to any one of embodiments 1 to 34, wherein the frequency of cells expressing IGF-1R in the subject is increased compared to cells from a healthy subject or a subject not diagnosed with the ocular inflammatory condition.

[0175] 37. The method according to embodiment 36, wherein the frequency of the IGF-1R-expressing cells is measured by flow cytometry or immunohistochemistry.

[0176] 38. The method according to any one of embodiments 1 to 37, wherein the treatment results in disease modification in the subject.

[0177] 39. The method according to any one of embodiments 1 to 38, wherein the IGF-1R ligand comprises wild-type insulin-like growth factor 1 (IGF-1) (SEQ ID NO: 3), wild-type insulin (SEQ ID NOs: 10 and 11), or wild-type insulin-like growth factor 2 (IGF-2) (SEQ ID NO: 12).

[0178] 40. The method according to any one of embodiments 1 to 38, wherein the IGF-1R ligand comprises a variant of wild-type IGF-1 (SEQ ID NO: 3), a variant of wild-type insulin (SEQ ID NOs: 10 and 11), or a variant of wild-type IGF-2 (SEQ ID NO: 12).

[0179] 41. The method according to embodiment 40, wherein the variant of wild-type IGF-1 is at least 90% identical to IGF-1 (SEQ ID NO: 3), the variant of wild-type insulin is at least 90% identical to insulin (SEQ ID NOs: 10 and 11), or the variant of wild-type IGF-2 is at least 90% identical to IGF-2 (SEQ ID NO: 12).

[0180] 42. The method according to embodiment 40 or 41, wherein (i) the variant of IGF-1 has a reduced binding affinity for insulin-like growth factor-binding protein IGF-1 (IGFBP) as compared to wild-type IGF-1 (SEQ ID NO: 3), or the variant of IGF-2 has a reduced binding affinity for IGFBP as compared to wild-type IGF-2 (SEQ ID NO: 12), and / or (ii) the variant of IGF-1 has an increased affinity for wild-type IGF-1R as compared to wild-type IGF-1 (SEQ ID NO: 3), or the variant of IGF-2 has an increased affinity for wild-type IGF-1R as compared to wild-type IGF-2 (SEQ ID NO: 12).

[0181] 43. The method according to any one of embodiments 1 to 38 or embodiments 40 to 42, wherein the IGF-1R ligand comprises 765IGF (SEQ ID NO: 2), IGF-132 (SEQ ID NO: 4), long-R3-IGF-1 (SEQ ID NO: 5), R3-IGF-1 (SEQ ID NO: 6), des(1-3)-IGF-1 (SEQ ID NO: 7), long-IGF-1 (SEQ ID NO: 8), or long-G3-IGF-1 (SEQ ID NO: 9).

[0182] 44. The method according to any one of embodiments 1 to 43, wherein the IGF-1R ligand, or a part or variant thereof, comprises a leader sequence.

[0183] 45. The method according to embodiment 44, wherein the leader sequence comprises SEQ ID NO: 1.

[0184] 46. The method according to any one of embodiments 43 to 45, wherein the IGF-1R ligand comprises 765IGF (SEQ ID NO: 2).

[0185] 47. The method according to any one of embodiments 1 to 46, wherein the disease modifier comprises a cytotoxic agent.

[0186] 48. The method according to embodiment 47, wherein the cytotoxic agent comprises a chemotherapeutic agent.

[0187] 49. The method according to embodiment 48, wherein the chemotherapeutic agent is amsacrine and is azacitidine, bleomycin, busulfan, capecitabine, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, cyclophosphamide, cytarabine, dactinomycin, daunorubicin, dacarbazine, docetaxel, doxorubicin, epirubicin, estramustine, etoposide, floxuridine, fludarabine, fluorouracil, gemcitabine, hexamethylmelamine, idarubicin, ifosfamide, irinotecan, lomustine, mechlorethamine, melphalan, mercaptopurine, methotrexate, mitomycin C, mitotane, mitoxantrone, oxaliplatin, paclitaxel, pemetrexed, pentostatin, plicamycin, procarbazine, raltitrexed, semustine, streptozocin, temozolomide, teniposide, thioguanine, thiotepa, topotecan, trimetrexate, valrubicin, vincristine, vinblastine, vindesine, or vinorelbine.

[0188] 50. The method according to embodiment 49, wherein the chemotherapeutic agent is methotrexate.

[0189] 51. The method according to embodiment 47, wherein the cytotoxic agent comprises a toxin.

[0190] 52. The method according to embodiment 51, wherein the toxin comprises Clostridium perfringens enterotoxin, diphtheria toxin, ricin chain A, Pseudomonas exotoxin, A-chain toxin, ribosome-inactivating protein, -sarcin, aspergillin, or ribonuclease.

[0191] 53. The method according to embodiment 52, wherein the toxin comprises Clostridium perfringens enterotoxin, or a part or variant thereof.

[0192] 54. The method according to embodiment 53, wherein the conjugate comprises SEQ ID NO: 14 or SEQ ID NO: 15.

[0193] 55. The method according to embodiment 52, wherein the toxin comprises diphtheria toxin, or a part or variant thereof.

[0194] 56. The method according to embodiment 55, wherein the conjugate comprises SEQ ID NO: 13 or SEQ ID NO: 16.

[0195] 57. The method according to any one of embodiments 1 to 46, wherein the disease modifier is selected from glucocorticoid (dexamethasone), corticosteroid (methylprednisolone, prednisone, triamcinolone acetonide), thyroid-stimulating hormone receptor (TSHR) inhibitor, mycophenolate mofetil, simvastatin, metformin, phenformin, cyclosporine, rapamycin, or another mammalian target agent of rapamycin (mTOR) inhibitor, or azathioprine.

[0196] 58. The method according to any one of embodiments 4 to 57, wherein the subject (i) has Graves' hyperthyroidism and / or (ii) has at least 3 or at least 4 CASs.

[0197] 59. The method according to any one of embodiments 1 to 58, wherein the subject (i) has not previously received treatment for the inflammatory condition of the eye, (ii) has previously received treatment for the inflammatory condition of the eye, (iii) has relapsed from a previous treatment for the inflammatory condition of the eye, or (iv) was refractory to a previous treatment for the inflammatory condition of the eye.

[0198] 60. The method according to embodiment 59, wherein the previous treatment comprises administration of an IGF-1R inhibitor to the subject.

[0199] 61. The method according to embodiment 60, wherein the IGF-1R inhibitor was an antibody.

[0200] 62. The method according to embodiment 61, wherein the antibody is teprotumumab, ganitumab, figitumumab, MEDI-573, siltuximab, daratumumab, robatumumab, AVE1642, BIIB022, xentuzumab, isothiazumab, lonidatumumab, VB421, VRDN-001, VRDN-002 or VRDN-003.

[0201] 63. The method according to embodiment 60, wherein the IGF-1R inhibitor is a small molecule.

[0202] 64. The method according to embodiment 63, wherein the small molecule is lapatinib, picropodophyllin, AXL1717, BMS-754807, BMS-536924, BMS-554417, GSK1838705A, GSK1904529A, NVP-AEW541, NVP-ADW742, GTx-134, AG1024, KW-2450, PL-2258, NVP-AEW541, NSM-18, AZD3463, AZD9362, BI885578, BI893923, TT-100, XL-228, or A-928605.

[0203] 65. The method according to embodiment 60, wherein the aforementioned IGF-1R inhibitor is a radioimmunoconjugate.

[0204] 66. The method according to embodiment 65, wherein the radioimmunoconjugate is FPI-1434.

[0205] 67. The method according to embodiment 60, wherein the aforementioned IGF-1R inhibitor is an antibody-drug conjugate.

[0206] 68. The method according to embodiment 67, wherein the antibody-drug conjugate is W0101.

[0207] 69. The method according to any one of embodiments 1 to 68, wherein the conjugate is administered in combination with one or more other therapies.

[0208] 70. The method according to embodiment 69, wherein the one or more other therapies are: glucocorticoids (dexamethasone), corticosteroids (methylprednisolone, prednisone, triamcinolone acetonide), rituximab or other anti-CD20 antibodies, tocilizumab, TOUR006, or other anti-IL-6 antibodies, selenium, infliximab, adalimumab, or other anti-tumor necrosis factor (TNF) α antibodies, TSHR inhibitors, orbital radiotherapy, vascular endothelial growth factor (VEGF) inhibitors, mycophenolate mofetil, isocablimab or other anti-CD40 antibodies, IMVT-1401, batoclimab, efgartigimod, or other neonatal fragment crystallizable receptor (FcRn) inhibitors, simvastatin, metformin, phenformin, cyclosporine, azathioprine, methotrexate, rapamycin or mTOR inhibitors, and / or intravenous immunoglobulin therapy.

[0209] 71. The method according to any one of embodiments 1-50 or embodiment 57, wherein the conjugate is administered at a dose of about 0.05, 0.10, 0.20, 0.40, 0.80, 1.0, 1.5, 1.6, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, or 10.0 μEq / kg body weight, or in a dose range of about 0.05-0.5, 0.5-1.0, 1.0-1.5, 1.5-2.0, 2.0-2.5, 2.5-3.0, 3.0-3.5, 3.5-4.0, 4.0-4.5, 4.5-5.0, 5.0-5.5, 5.5-6.0, 6.0-6.5, 6.5-7.0, 7.0-7.5, 7.5-8.0, 8.0-8.5, 8.5-9.0, 9.0-9.5, or 9.5-10.0 μEq / kg body weight.

[0210] 72. The conjugate is administered at a dose of about 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13.0, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14.0, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, 15.0, 15.1, 15.2, 15.3, 15.4, 15.5, 15.6, 15.7, 15.8, 15.9, 16.0, 16.1, 16.2, 16.3, 16.4, or 16.5 mg / kg body weight, or in the range of about 0.05 - 0.5, 0.5 - 1.0, 1.0 - 1.5, 1.5 - 2.0, 2.0 - 2.5, 2.5 - 3.0, 3.0 - 3.5, 3.5 - 4.0, 4.0 - 4.5, 4.5 - 5.0, 5.0 - 5.5, 5.5 - 6.0, 6.0 - 6.5, 6.5 - 7.0, 7.0 - 7.5, 7.5 - 8.0, 8.0 - 8.5, 8.5 - 9.0, 9.0 - 9.5, 9.5 - 10.0, 10.0 - 10.5, 10.5 - 11.0, 11.0 - 11.5, 11.5 - 12.0, 12.0 - 12.5, 12.5 - 13.0, 13.0 - 13.5, 13.5 - 14.0, 14.0 - 14.5, 14.5 - 15.0, 15.0 - 15.5, 15.5 - 16.0, or in a dosage range of 16.0 - 16.5 mg / kg body weight, the method according to any one of Embodiments 1 - 71.

[0211] 73. The method according to any one of Embodiments 1 - 70, wherein the conjugate is administered at a dosage that is the maximum tolerated dose.

[0212] 74. The method according to any one of Embodiments 1 - 73, wherein the conjugate is administered once daily, every other day, once every 3 days, once every 4 days, once every 5 days, once every 6 days, once a week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once a month, once every 2 months, or once every 3 months.

[0213] 75. The method according to any one of Embodiments 1 - 74, wherein the conjugate is administered via intravenous, subcutaneous, intravitreal injection, via retrobulbar injection, or via an eye dropper.

[0214] 76. The method according to any one of Embodiments 1 - 75, wherein the method does not cause unacceptable hyperglycemia in the subject.

[0215] 77. A method for treating TED in a subject, the method comprising administering to the subject an effective amount of a conjugate, the conjugate comprising (i) 765IGF (SEQ ID NO: 2), and (ii) methotrexate.

[0216] The following examples are provided for illustrative purposes and not for purposes of limitation.

Example

[0217] Example 1: Evaluation of the Efficacy of LX - 101 on TED Orbital Fibroblasts LX-101 is an IGF-1R ligand (SEQ ID NO: 2) conjugated to methotrexate, a well-known inhibitor of dihydrofolate reductase, which ultimately inhibits nucleotide synthesis. Without wishing to be bound by theory, LX-101 may deliver methotrexate to cells that overexpress IGF-1R involved in the pathology of TED, reducing their survival rate and proliferation. Primary orbital fibroblasts (OF) are obtained from patients with TED who have undergone orbital decompression surgery and are cultured with LX-101 in a dose range of up to 2500 nM (expressed with respect to IGF-1 protein). For comparison, some OF are treated with IGF-1 to activate normal IGF-1R signaling. The survival rate is determined using an alamar blue assay or any survival assay known in the art. Proliferation is determined using BrdU incorporation or any proliferation assay known in the art.

[0218] OF can be treated with cytokines for activation, including platelet-derived growth factor BB (PDGF-BB) that stimulates OF proliferation, and transforming growth factor β (TGF-β) that promotes OF differentiation into myofibroblasts and scar tissue formation. Additional activators of OF include IL-1, IL-4, IL-6, IL-17, leukoregulin, IGF-1, TNF-α, IFN-γ, CD40L, PPARγ agonist, and autoantibodies from patients with Graves' disease (e.g., autoantibodies against TSHR). The effect of LX-101 on activated OF is studied. Without wishing to be bound by theory, LX-101 may have a more significant effect on activated OF, which is related to the pathology of TED.

[0219] For one cell viability assay, OF is seeded in a black 96-well plate with appropriate treatment and culture medium. Puromycin is used as a cytotoxic positive control. The cells are incubated with the treatment for a certain period and then Alamar Blue reagent is added to each well. After incubation, the fluorescence of the oxidized Alamar reagent is measured (excitation 470 nm, emission 480 nm) and normalized to vehicle-treated cells.

[0220] For one cell proliferation assay, OF strains are seeded in 96-well plates. The cells are treated three times with a specific treatment for a certain period (e.g., 2 - 14 days) before adding bromodeoxyuridine (BrdU) for 18 - 24 hours, and proliferation is measured using a BrdU cell proliferation assay kit. The samples are fixed, stained with anti-BrdU antibody, incubated with the corresponding horseradish peroxidase (HRP)-conjugated secondary antibody, and evaluated using a microplate reader.

[0221] Without wishing to be bound by theory, LX-101 may reduce the production of hyaluronic acid (HA) produced by OF, which contributes to the pathology of TED. To detect and analyze HA production, after treatment with LX-101, the supernatant is collected from OF and the amount of HA in each sample is measured by ELISA.

[0222] Interleukin-6 (IL-6), a pro-inflammatory cytokine produced by lymphocytes, monocytes, and OF, is present at high concentrations in TED patients and plays an important role in the etiology of the disease. Without wishing to be bound by theory, LX-101 may reduce the production of IL-6. To detect and analyze IL-6 production, after treatment with LX-101, the supernatant is collected from OF and the amount of IL-6 in each sample is measured by ELISA.

[0223] Although not wishing to be bound by theory, LX-101 may inhibit the inflammatory phenotype of OF. ELISA and Western blot can be used to detect and analyze the molecular mediators of inflammation (e.g., COX-2).

[0224] The effects of LX-101 in OFs not derived from TED and / or fibroblasts from other tissues (e.g., eyelid or dermal fibroblasts) can also be studied and compared.

[0225] Since TED is a complex disease involving various cells, study the effect of LX-101 on OFs grown in co-culture with immune cells. Lymphocytes play an important role in the early stages of TED and can initiate the inflammatory process and cytokine production. Grow OFs in co-culture with B cells and / or T cells from TED patients and study the effect of LX-101 on OF survival, proliferation, and phenotype.

[0226] Example 2: Investigation of the mechanism of action of LX-101 in TED OF Culture primary OFs from patients with TED and treat them with various doses of LX-101 (e.g., 2500 - 1.64 nM expressed as IGF-1 protein) for a defined period (e.g., 2 - 14 days). The specificity of LX-101 targeting IGF-1R can be studied by treating OFs with siRNA specific to IGF-1R to deplete IGF-1R or using a non-specific siRNA control. Although not wishing to be bound by theory, the effect of LX-101 on OFs may be abrogated by depletion of IGF-1R expression.

[0227] The binding affinity of LX-101 to IGF-IR on TED OF and its competition with endogenous ligands such as IGF-1 are characterized. The effect of LX-101 on the IGF-1R signaling pathway is determined, for example, by Western blot by detecting the phosphorylation of intracellular signaling proteins such as PI3K, AKT, MAPK, and JNK. The specificity of LX-101 targeting IGF-1R can be determined by depleting IGF-1R as described above. Furthermore, it is determined whether LX-101 induces apoptosis in TED OF.

[0228] Example 3: Investigate the effect of LX-101 on adipocytes and myofibroblasts different from TED OF Primary OF from patients with TED are treated with various doses of LX-101 (e.g., 2500 - 1.64 nM, expressed with respect to the IGF-1 protein) for a certain period (e.g., 2 - 14 days) and further cultured to differentiate into myofibroblasts or adipocytes. OF is cultured to differentiate into myofibroblasts by the addition of TGF-β regardless of the presence or absence of the addition of IGF-1. The effects of myofibroblast formation and activation in the presence of LX-101 are measured by Western blot, and RT-qPCR (e.g., for targets such as smooth muscle actin, collagen, and fibronectin), immunofluorescence (e.g., for detection of HA accumulation of actin filament formation), and ELISA (HA production).

[0229] Furthermore, OF is cultured and differentiated into adipocytes using a standard adipogenic medium. Adipocyte formation is measured by Western blot, RT-qPCR, and immunofluorescence of adipogenic markers (e.g., FABP4, PPARγ, lipid accumulation). The effect of LX-101 on OF from patients who did not respond to teprotumumab is also tested. Thus, the role of LX-101 targeting myofibroblast and adipocyte differentiation is investigated. Without wishing to be bound by theory, LX-101 may inhibit the ability of OF to differentiate into myofibroblasts and adipocytes and may directly inhibit the survival of differentiated myofibroblasts and adipocytes.

[0230] Example 4: Investigate the effect of LX-101 on the survival of circulating PBMCs from TED patients Peripheral blood mononuclear cells (PBMCs) from patients with TED are obtained and treated with various doses of LX-101 (e.g., 2500 - 1.64 nM, expressed with respect to IGF-1 protein) for a certain period (e.g., 2 - 14 days) and evaluated by flow cytometry. Cells are identified by CD markers (e.g., CD34 for fibroblasts, CD20 for B cells, CD3 for T cells, CD14 for monocytes). Cell survival is measured by 7-AAD staining or any method known in the art, and proliferation is measured by BrdU labeling or any method known in the art.

[0231] In some PBMC cultures, T cells are activated using anti-CD28 / CD3 magnetic beads and recombinant IL-2. The expanded T cells are treated with IGF-1 + / - LX-101, and survival and proliferation are measured. The ability of LX-101 to target additional cell types involved in the pathophysiology of TED is determined. Without wishing to be bound by theory, LX-101 may inhibit the survival and / or proliferation of fibroblasts, T cells, and B cells from TED patients. Furthermore, it is determined whether LX-101 targets IGF-1 signaling in T cells from TED patients.

[0232] Efforts have been made to ensure accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.), but some experimental errors and deviations should be taken into account.

[0233] One of ordinary skill in the art will recognize many methods and materials similar or equivalent to those described herein that can be used in practicing the subject matter described herein. The present disclosure is not limited to the methods and materials described.

[0234] Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this subject matter belongs.

[0235] Throughout this specification and the claims, the words "comprise", "comprises", and "comprising" are used in a non-exclusive sense, unless the context requires otherwise. It is understood that the embodiments described herein include embodiments consisting of and / or consisting essentially of the embodiments.

[0236] When a range of values is provided, each intervening value, to the tenth of the unit of the lower limit, is understood to be included between the upper and lower limits of the range, and any other stated value or intervening value within the stated range, unless the context clearly dictates otherwise. Upper and lower limits of these smaller ranges that may independently be included in a smaller range are also included, subject to any specifically excluded limit within the stated range. When the stated range includes one or both of the limits, ranges excluding one or both of those included limits are also included.

[0237] Many modifications of the invention described herein, and other embodiments, will come to mind to those of ordinary skill in the art to which this invention pertains, having the benefit of the teachings presented in the foregoing description and the related drawings. Accordingly, it is to be understood that the invention is not to be limited to the specific embodiments disclosed, and that modifications and other embodiments are intended to be included within the scope of the appended claims. Specific terms are used herein, but they are used in a generic and descriptive sense only and not for purposes of limitation.

Claims

**Claim 1** A method of treating an inflammatory condition of the eye in a subject in need thereof, the method comprising administering to the subject a conjugate comprising an insulin-like growth factor 1 receptor (IGF-1R) ligand, or a portion or variant thereof, and a disease modifying agent. **Claim 2** The method of claim 1, wherein the inflammatory condition of the eye is selected from the group consisting of thyroid eye disease (TED), uveitis, scleritis, keratitis, conjunctivitis, and orbital inflammatory disease. **Claim 3** The method of claim 2, wherein the orbital inflammatory disease is selected from the group consisting of idiopathic orbital inflammation, orbital inflammatory pseudotumor, orbital myositis, inflammatory orbital cellulitis, optic nerve sheath meningitis, episcleritis, diffuse orbital inflammation, orbital apex syndrome, or sclerosing orbital inflammation. **Claim 4** The method of claim 2, wherein the inflammatory condition of the eye is TED. **Claim 5** The method of claim 4, wherein the TED is selected from the group consisting of active TED, acute TED, inactive TED, chronic TED, moderate to severe TED, or vision-threatening TED. **Claim 6** The method of claim 4 or 5, wherein the method results in a reduction of proptosis in the subject. **Claim 7** The method of claim 6, wherein the proptosis is reduced by at least 2 mm, at least 3 mm, or at least 4 mm. **Claim 8** The method of claim 6 or 7, wherein the reduction of proptosis is evaluated by a proptometer or orbital imaging. **Claim 9** The method of claim 8, wherein the orbital imaging is a computed tomography (CT) scan or magnetic resonance imaging. **Claim 10** The reduction of proptosis is associated with a reduction in the amount of extraocular muscle. The method according to any one of claims 6 to 9. **Claim 11** The reduction of proptosis is associated with a reduction in orbital fat volume. The method according to any one of claims 6 to 9. **Claim 12** The method of any one of claims 4 to 11, wherein the method results in a reduction of clinical activity score (CAS) in the subject. **Claim 13** The method of claim 12, wherein the CAS is reduced by at least 2 points or at least 3 points. **Claim 14** The method of claim 12, wherein the CAS is reduced to 1 or reduced to 0. **Claim 15** The method of any one of claims 4 to 14, wherein the method results in a reduction of the severity of diplopia in the subject. **Claim 16** The method according to claim 15, wherein the reduction in the severity of diplopia is measured by the Gorman subjective diplopia score.

17. The method according to claim 15 or 16, wherein the diplopia is selected from the group consisting of constant diplopia, intermittent diplopia, or indeterminate diplopia.

18. The method according to any one of claims 15 to 17, wherein the method results in the complete disappearance of diplopia.

19. The method according to any one of claims 16 to 18, wherein the severity of diplopia in the subject is reduced by at least one grade.

20. The method according to any one of claims 4 to 19, wherein the method results in an improvement in the quality of life of the subject.

21. The method according to claim 20, wherein the improvement in the quality of life is measured by the Graves' ophthalmopathy quality of life (GO-QoL) assessment scale.

22. The method according to claim 20 or 21, wherein the quality of life is measured by the visual function subscale of GO-QoL, or the appearance subscale of GO-QoL.

23. The method according to claim 21 or 22, wherein the quality of life is improved by at least 8 points.

24. The method according to any one of claims 4 to 23, wherein the method results in an improvement in neuropathy.

25. The method according to any one of claims 4 to 24, wherein the method results in a reduction in retro-orbital edema.

26. The method according to any one of claims 4 to 25, wherein the method results in an improvement in monocular retraction movement.

27. The method according to claim 26, wherein the improvement in monocular retraction movement is measured by a light reflex test.

28. The method according to claim 27, wherein the improvement is at least 10 degrees.

29. The method according to any one of claims 1 to 28, wherein the subject has a decrease in the number of IGF-1R-expressing cells.

30. The method according to claim 29, wherein the decrease in the number of IGF-1R-expressing cells is measured by flow cytometry, or immunohistochemistry.

31. The method according to claim 29 or 30, wherein the IGF-1R-expressing cells are orbital fibroblasts (OF).

32. The method according to any one of claims 4 to 31, wherein the method results in a reduction or inhibition of hyaluronan synthesis in the posterior chamber of the eye.

33. The method according to any one of claims 4 to 32, wherein the method results in a reduction or inhibition of adipogenesis or a reduction of adipocytes in the posterior chamber of the eye of the subject.

34. The method according to any one of claims 4 to 33, wherein the method results in a reduction in the levels of interleukin (IL)-6, IL-16, and / or RANTES in the serum of the subject.

35. The method according to claim 29 or 30, wherein the IGF-1R-expressing cells are selected from the group consisting of fibroblasts, B lymphocytes, and T lymphocytes.

36. The method according to any one of claims 1 to 35, wherein the IGF-1R ligand comprises wild-type insulin-like growth factor 1 (IGF-1), wild-type insulin, or wild-type insulin-like growth factor 2 (IGF-2).

37. The method according to claim 36, wherein the wild-type IGF-1 comprises SEQ ID NO: 3, the wild-type insulin comprises SEQ ID NO: 10 or 11, and the wild-type IGF-2 comprises SEQ ID NO:

12.

38. The method according to any one of claims 1 to 37, wherein the IGF-1R ligand comprises a variant of wild-type IGF-1, a variant of wild-type insulin, or a variant of wild-type IGF-2.

39. The method according to claim 38, wherein the variant of wild-type IGF-1 is at least 90% identical to SEQ ID NO: 3, the variant of wild-type insulin is at least 90% identical to SEQ ID NO: 10 or 11, and the variant of wild-type IGF-2 is at least 90% identical to SEQ ID NO:

12.

40. The method according to claim 38 or 39, wherein (i) the variant of wild-type IGF-1 has a reduced binding affinity for insulin-like growth factor binding protein (IGFBP) compared to wild-type IGF-1, or the variant of wild-type IGF-2 has a reduced binding affinity for IGFBP compared to wild-type IGF-2, and / or (ii) the variant of wild-type IGF-1 has an increased affinity for IGF-1R compared to wild-type IGF-1, or the variant of wild-type IGF-2 has an increased affinity for IGF-1R compared to wild-type IGF-2.

41. The method according to any one of claims 1 to 40, wherein the IGF-1R ligand, or a part or variant thereof, comprises a leader sequence.

42. The method according to claim 41, wherein the leader sequence comprises SEQ ID NO:

1.

43. The method according to any one of claims 1 to 35 and claims 38 to 42, wherein the IGF-1R ligand comprises 765IGF (SEQ ID NO: 2), IGF-132 (SEQ ID NO: 4), long-R3-IGF-1 (SEQ ID NO: 5), R3-IGF-1 (SEQ ID NO: 6), des(1-3)-IGF-1 (SEQ ID NO: 7), long-IGF-1 (SEQ ID NO: 8), or long-G3-IGF-1 (SEQ ID NO: 9).

44. The method according to claim 43, wherein the IGF-1R ligand comprises 765IGF (SEQ ID NO: 2).

45. The method according to any one of claims 1 to 44, wherein the IGF-1R ligand, or a part or variant thereof, is covalently bound to the disease modifier, and the disease modifier comprises a cytotoxic agent.

46. The method according to claim 45, wherein the cytotoxic agent comprises a chemotherapeutic agent.

47. The method according to claim 46, wherein the chemotherapeutic agent is amsacrine, azacitidine, bleomycin, busulfan, capecitabine, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, cyclophosphamide, cytarabine, dactinomycin, daunorubicin, dacarbazine, docetaxel, doxorubicin, epirubicin, estramustine, etoposide, floxuridine, fludarabine, fluorouracil, gemcitabine, hexamethylmelamine, idarubicin, ifosfamide, irinotecan, lomustine, mechlorethamine, melphalan, mercaptopurine, methotrexate, mitomycin C, mitotane, mitoxantrone, oxaliplatin, paclitaxel, pemetrexed, pentostatin, plicamycin, procarbazine, raltitrexed, semustine, streptozocin, temozolomide, teniposide, thioguanine, thiotepa, topotecan, trimetrexate, valrubicin, vincristine, vinblastine, vindesine, or vinorelbine.

48. The method according to claim 47, wherein the chemotherapeutic agent is methotrexate.

49. The method according to claim 45, wherein the cytotoxic agent comprises a toxin.

50. The method according to claim 49, wherein the toxin comprises Clostridium perfringens enterotoxin, diphtheria toxin, ricin chain A, Pseudomonas exotoxin, A chain toxin, ribosome-inactivating protein, α-sarcin, aspergillin, or ribonuclease.

51. The method according to claim 50, wherein the toxin comprises Clostridium perfringens enterotoxin, or a part or variant thereof.

52. The method according to claim 51, wherein the conjugate comprises SEQ ID NO: 14 or SEQ ID NO:

15.

53. The method according to claim 50, wherein the toxin comprises diphtheria toxin, or a part or variant thereof.

54. The method according to claim 53, wherein the toxin comprises SEQ ID NO: 13 or SEQ ID NO:

16.

55. The method according to any one of claims 1 to 44, wherein the disease modifier is selected from the group consisting of glucocorticoid, corticosteroid, thyroid-stimulating hormone receptor (TSHR) inhibitor, mycophenolate mofetil, simvastatin, metformin, phenformin, cyclosporine, rapamycin, a mammalian target of rapamycin (mTOR) inhibitor, and azathioprine.

56. The method according to any one of claims 1 to 55, wherein the conjugate is administered at a dose of about 0.05, 0.10, 0.20, 0.40, 0.80, 1.0, 1.5, 1.6, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, or 10.0 μeq / kg body weight, or in a dose range of about 0.05 - 0.5, 0.5 - 1.0, 1.0 - 1.5, 1.5 - 2.0, 2.0 - 2.5, 2.5 - 3.0, 3.0 - 3.5, 3.5 - 4.0, 4.0 - 4.5, 4.5 - 5.0, 5.0 - 5.5, 5.5 - 6.0, 6.0 - 6.5, 6.5 - 7.0, 7.0 - 7.5, 7.5 - 8.0, 8.0 - 8.5, 8.5 - 9.0, 9.0 - 9.5, or 9.5 - 10.0 μeq / kg body weight.

57. The conjugate is in a dosage range of about 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13.0, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14.0, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, 15.0, 15.1, 15.2, 15.3, 15.4, 15.5, 15.6, 15.7, 15.8, 15.9, 16.0, 16.1, 16.2, 16.3, 16.4, or 16.5 mg / kg body weight, or in a dosage range of about 0.05 - 0.5, 0.5 - 1.0, 1.0 - 1.5, 1.5 - 2.0, 2.0 - 2.5, 2.5 - 3.0, 3.0 - 3.5, 3.5 - 4.0, 4.0 - 4.5, 4.5 - 5.0, 5.0 - 5.5, 5.5 - 6.0, 6.0 - 6.5, 6.5 - 7.0, 7.0 - 7.5, 7.5 - 8.0, 8.0 - 8.5, 8.5 - 9.0, 9.0 - 9.5, 9.5 - 10.0, 10.0 - 10.5, 10.5 - 11.0,The method according to any one of claims 1 to 56, which is administered to a subject in a dosage range of 11.0 to 11.5, 11.5 to 12.0, 12.0 to 12.5, 12.5 to 13.0, 13.0 to 13.5, 13.5 to 14.0, 14.0 to 14.5, 14.5 to 15.0, 15.0 to 15.5, 15.5 to 16.0, or 16.0 to 16.5 mg / kg body weight.

58. The method according to any one of claims 1 to 57, wherein the conjugate is administered at a dose that is the maximum tolerated dose.

59. The method according to any one of claims 1 to 58, wherein the conjugate is administered daily, once every two days, once every three days, once every four days, once every five days, once every six days, once a week, once every two weeks, once every three weeks, once every four weeks, once a month, once every two months, or once every three months.

60. The method according to any one of claims 1 to 59, wherein the conjugate is administered via intravenous, subcutaneous, intravitreal injection, via retrobulbar injection, or via an eye dropper.

61. The method according to any one of claims 1 to 35 and 38 to 60, wherein the IGF-1R ligand is SEQ ID NO: 2, the cytotoxic agent is methotrexate, there are 6 to 10 methotrexate molecules per IGF-1R ligand of SEQ ID NO: 2, and the ocular inflammatory condition is TED.

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