Treatment of autoimmune diseases with insulin-like growth factor 1 receptor ligands conjugated to agents

Abstract

The subject matter described herein provides a method for treating an autoimmune disease 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 alleviator.

Description

[0001] Cross-reference to related applications

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 367,240, filed on June 29, 2022. Technical Field

[0003] The presently disclosed subject matter is generally directed to methods of treating autoimmune diseases, particularly by administering an IGF-1R ligand conjugated to a cytotoxic agent.

[0004] Reference to a sequence listing

[0005] The sequence listing written into the file name 597947SEQLIST.xml is 17 kilobytes, created on June 29, 2023, and is hereby incorporated by reference. Background Art

[0006] The human immune system is an intricate network of different cell types that has evolved to protect the host from pathogens and malignancies, and to maintain tissue homeostasis. Autoimmune diseases occur when the immune system is misdirected to host tissues, usually due to loss of B-cell or T-cell tolerance, and can range from organ-specific diseases to systemic diseases. Despite recent advances in research, autoimmune diseases remain poorly understood, are difficult to treat, and contribute significantly to annual morbidity and mortality.

[0007] There remains a need in the art for improved methods of treating autoimmune diseases. Summary of the invention

[0008] In accordance with this specification, the present application describes methods of treatment with an IGF-1R ligand conjugated to a disease modifying agent.

[0009] In one embodiment, the present application provides a method for treating an autoimmune disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a conjugate, wherein the conjugate comprises an IGF-1R ligand and a disease-modifying agent.

[0010] In some embodiments, the autoimmune disease is selected from the group consisting of: adrenergic drug resistance, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, adrenal autoimmune disease, allergic encephalomyelitis, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inflammatory eye disease, autoimmune neonatal thrombocytopenia, autoimmune neutropenia, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, autoimmune thyroiditis, Behcet's disease, bullous pemphigoid, cardiomyopathy, cardiotomy syndrome, celiac disease-dermatitis, chronic active hepatitis, chronic fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, Churg-Strauss syndrome (Churg-Strauss syndrome), syndrome), cicatricial pemphigoid, CREST syndrome, cold agglutinin disease, Crohn's disease, Cushing's syndrome, cutaneous graft-versus-host disease (GVHD), dense deposit disease, dermatomyositis, essential mixed cryoglobulinemia, fibromyalgia-fibromyositis, glomerulonephritis (e.g., IgA nephropathy), gluten-sensitive enteropathy, Goodpasture's syndrome, Graves' disease, Guillain-Barre disease, hyperthyroidism (e.g., Hashimoto's thyroiditis), idiopathic pulmonary fibrosis, idiopathic Addison's disease, idiopathic thrombocytopenic purpura (ITP), IgA neuropathy, inflammatory arthritis, irritable bowel disease, juvenile arthritis, lichen planus, lichen sclerosus, lupus [e.g.,Systemic lupus erythematosus (SLE), cutaneous lupus, discoid lupus, Ménière's disease, mixed connective tissue disease, morphea, multiple sclerosis, myasthenia gravis, myocarditis, type 1 or immune-mediated diabetes mellitus, neuritis, other endocrine failure, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychondritis, polyendocrinopathy, polyglandular syndrome, polymyalgia rheumatica, polymyositis, post-MI complications, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud's phenomenon, relapsing polychondritis, Reiter's syndrome, rheumatic heart disease, rheumatoid arthritis, sarcoidosis, Sjogren's syndrome, stiff-person syndrome, systemic sclerosis (SSc), Takayasu's arteritis arteritis), temporal arteritis / giant cell arteritis, ulcerative colitis, urticaria, uveitis, uveitic ophthalmia, vasculitis such as dermatitis herpetiformis, vitiligo, and Wegener's granulomatosis.

[0011] In one embodiment, the autoimmune disease is rheumatoid arthritis, Graves' disease, SSc, Cushing's syndrome, idiopathic pulmonary fibrosis, SLE or Crohn's disease. In another embodiment, the SSc is diffuse cutaneous SSc.

[0012] In some embodiments, the treatment results in functional changes or damage to the cells expressing IGF-1R of the subject. In other embodiments, the treatment reduces the number of cells expressing IGF-1R of the subject. In a specific embodiment, the reduction is caused by killing of cells expressing IGF-1R. Without being bound by theory, it is believed that simple inhibition of the IGF-1R pathway may be insufficient as a completely curative treatment due to signal transduction through other potentially redundant pathways, or compensatory mechanisms, or pathways that are upregulated and may lead to adaptive resistance that may continue to act as disease effectors. Therefore, targeted delivery of disease-modifying agents (including agents that cause killing of cells expressing IGF-1R) can provide benefits beyond (including superior to) simple IGF-1R pathway inhibition because it bypasses the effects of all such potentially redundant and / or compensatory pathways.

[0013] In another specific embodiment, the cells expressing IGF-1R are B lymphocytes and / or T lymphocytes.

[0014] In another embodiment, IGF-1R is overexpressed by cells in the subject relative to its expression by cells from a healthy subject or a subject not diagnosed as having an autoimmune disease.

[0015] In another embodiment, the frequency of cells expressing IGF-1R in the subject is increased relative to the frequency of cells from a healthy subject or a subject not diagnosed with an autoimmune disease. In another embodiment, the frequency of cells expressing IGF-1R is measured by flow cytometry or immunohistochemistry.

[0016] In some embodiments, said treatment results in disease remission in said subject.

[0017] In some embodiments, the conjugate comprises wild-type insulin-like growth factor 1 (IGF-1) (SEQ ID NO: 3), wild-type insulin (SEQ ID NO: 10 and SEQ ID NO: 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 NO: 10 and SEQ ID NO: 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 NO: 10 and SEQ ID NO: 11), or the variant of wild-type IGF-2 is at least 90% identical to IGF-2 (SEQ ID NO: 12).

[0018] In some embodiments, the conjugate comprises a variant of IGF-1 that has reduced binding affinity for an IGF binding protein (IGFBP) compared to the binding affinity of wild-type IGF-1 (SEQ ID NO: 3) to the IGF binding protein, or a variant of IGF-2 that has reduced binding affinity for an IGF binding protein compared to the binding affinity of wild-type IGF-2 (SEQ ID NO: 12) to the IGF binding protein. In some embodiments, the variant of IGF-1 has increased affinity for the IGF-1R compared to the affinity of wild-type IGF-1 for the IGF-1R, or the variant of IGF-2 has increased affinity for the IGF-1R compared to the affinity of wild-type IGF-2 for the IGF-1R. In a specific embodiment, 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).

[0019] In some embodiments, the IGF-1R ligand, or a portion or variant thereof, comprises a leader sequence. In a specific embodiment, the leader sequence comprises SEQ ID NO:1.

[0020] In a specific embodiment, the conjugate comprises 765IGF (SEQ ID NO: 2).

[0021] In some embodiments, the disease-modifying agent comprises a cytotoxic agent. In some embodiments, the cytotoxic agent comprises a chemotherapeutic agent. In a specific embodiment, the chemotherapeutic agent is amuscarine, azacytidine, bleomycin, busulfan, capecitabine, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, cyclophosphamide, cytarabine, dactinomycin, daunorubicin, dacarbazine, docetaxel, doxorubicin, in), epirubicin, estramustine, etoposide, floxuridine, fludarabine, fluorouracil, gemcitabine, hexamethylmelamine, idarubicin, ifosfamide, irinotecan, lomustine, mechlorethamine, melphalan, mercaptopurine, methotrexate, mitomycin CIn another specific embodiment, the chemotherapeutic agent is methotrexate.

[0022] In some embodiments, the cytotoxic agent comprises a toxin. In a specific embodiment, 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, aspergillus, abrin, restrictomycin, bacterial endotoxin, lipid A portion of bacterial endotoxin, cholera toxin or ribonuclease. In a specific embodiment, the toxin comprises a portion of Clostridium perfringens enterotoxin or a variant thereof. In a specific embodiment, the conjugate comprises SEQ ID NO: 14 or SEQ ID NO: 15. In other specific embodiments, the toxin comprises diphtheria toxin or a portion or variant thereof. In a specific embodiment, the conjugate comprises SEQ ID NO: 13 or SEQ ID NO: 16.

[0023] In some embodiments, the subject having an autoimmune disease treated according to the methods described herein has not previously received treatment for the autoimmune disease, has previously received treatment for the autoimmune disease, has relapsed from a previous treatment for the autoimmune disease, or is refractory to a previous treatment for the autoimmune disease.

[0024] In some embodiments, the conjugate is administered in combination with one or more other therapies. In a specific embodiment, the one or more other therapies include one or more of the following: prednisone, hydroxychloroquine, chloroquine, belimumab, anifrolumab, abatacept, atacicept, lupuzor, rituximab, voclosporin, aldesleukin, baricitinib, BIIB059, BI655064, bortezomib, BT063, cenerimod, dapirolizumabpegol), edratide, filgotinib, GS-9876, iberdomide, IFN-α kinin, iguratimod, nelfinavir, obinutuzumab, OMS721, rapamycin, RC18, RSLV-132, SM101, theralizumab, ustekinumab ), vobarilizumab, XmAb5871, blisibimod, tabalumab, epratuzumab, rigerimod, tacrolimus, rontalizumab, sifalimumab, anifrolumab, tocilizumab, infliximab iximab), metelimumab, fresolimumab, rilonacept, cyclophosphamide, methotrexate, nintedanib, JBT-101, imatinib, pirfenidone, nilotinib, dasatinib, SAR100842, BMS-986202, BAY41-2272, riociguat guat), resunab, ixekizumab, brodalumab, tralokinumab, etanercept, adalimumab, golimumab, secukinumab, tildrakizumab, tofacitinib, and guselkumab.

[0025] In some embodiments, the conjugate is administered at about 0.05 μEq / kg body weight, 0.10 μEq / kg body weight, 0.20 μEq / kg body weight, 0.40 μEq / kg body weight, 0.80 μEq / kg body weight, 1.0 μEq / kg body weight, 1.5 μEq / kg body weight, 1.6 μEq / kg body weight, 2.0 μEq / kg body weight, 2.5 μEq / kg body weight, 3.0 μEq / kg body weight, 3.5 μEq / kg body weight. , 4.0 μEq / kg body weight, 4.5 μEq / kg body weight, 5.0 μEq / kg body weight, 5.5 μEq / kg body weight, 6.0 μEq / kg body weight, 6.5 μEq / kg body weight, 7.0 μEq / kg body weight, 7.5 μEq / kg body weight, 8.0 μEq / kg body weight, 8.5 μEq / kg body weight, 9.0 μEq / kg body weight, 9.5 μEq / kg body weight or 10.0 μEq / kg body weight or about 0. 05-0.5μEq / kg body weight, 0.5-1.0μEq / kg body weight, 1.0-1.5μEq / kg body weight, 1.5-2.0μEq / kg body weight, 2.0-2.5μEq / kg body weight, 2. 5-3.0μEq / kg body weight, 3.0-3.5μEq / kg body weight, 3.5-4.0μEq / kg body weight, 4.0-4.5μEq / kg body weight, 4.5-5.0μEq / kg body weight, 5.0- A dose range of 5.5 μEq / kg body weight, 5.5-6.0 μEq / kg body weight, 6.0-6.5 μEq / kg body weight, 6.5-7.0 μEq / kg body weight, 7.0-7.5 μEq / kg body weight, 7.5-8.0 μEq / kg body weight, 8.0-8.5 μEq / kg body weight, 8.5-9.0 μEq / kg body weight, 9.0-9.5 μEq / kg body weight or 9.5-10.0 μEq / kg body weight is administered.

[0026] In some embodiments, the conjugate is administered at about 0.05 mg / kg body weight, 0.10 mg / kg body weight, 0.15 mg / kg body weight, 0.20 mg / kg body weight, 0.25 mg / kg body weight, 0.30 mg / kg body weight, 0.35 mg / kg body weight, 0.40 mg / kg body weight, 0.45 mg / kg body weight, 0.50 mg / kg body weight, 0.55 mg / kg body weight, 0.60 mg / kg body weight, 0.65 mg / kg body weight, 0.70 mg / kg body weight, 0.75 mg / kg body weight, 0.80 mg / kg body weight, 0.85 mg / kg body weight, 0.90 mg / kg body weight, 0.95 mg / kg body weight, 1.0 mg / kg body weight / kg body weight, 1.1 mg / kg body weight, 1.2 mg / kg body weight, 1.3 mg / kg body weight, 1.4 mg / kg body weight, 1.5 mg / kg body weight, 1.6 mg / kg body weight, 1.7 mg / kg body weight, 1.8 mg / kg body weight, 1.9 mg / kg body weight, 2.0 mg / kg body weight, 2.1 mg / kg Body weight, 2.2 mg / kg body weight, 2.3 mg / kg body weight, 2.4 mg / kg body weight, 2.5 mg / kg body weight, 2.6 mg / kg body weight, 2.7 mg / kg body weight, 2.8 mg / kg body weight, 2.9 mg / kg body weight, 3.0 mg / kg body weight, 3.1 mg / kg body weight, 3.2 mg / kg body weight, 3.3 mg / kg body weight, 3.4 mg / kg body weight, 3.5 mg / kg body weight, 3.6 mg / kg body weight, 3.7 mg / kg body weight, 3.8 mg / kg body weight, 3.9 mg / kg body weight, 4.0 mg / kg body weight, 4.1 mg / kg body weight, 4.2 mg / kg body weight, 4.3 mg / kg body weight, 4.4 mg / kg body weight, 4.5 mg / kg body weight, 4.6 mg / kg body weight, 4.7 mg / kg body weight, 4.8 mg / kg body weight, 4.9 mg / kg body weight, 5.0 mg / kg body weight, 5.1 mg / kg body weight, 5.2 mg / kg body weight, 5.3 mg / kg body weight, 5.4 mg / kg body weight, 5.5 mg / kg body weight, 5.6 mg / kg body weight, 5.7 mg / kg body weight, 5.8 mg / kg body weight, 5.9 mg / kg body weight, 6.0 mg / kg body weight, 6.1 mg / kg body weight, 6.2 mg / kg body weight, 6.3 mg / kg body weight, 6.4 mg / kg body weight, 6.5 mg / kg body weight, 6.6 mg / kg body weight Weight, 6.7 mg / kg body weight, 6.8 mg / kg body weight, 6.9 mg / kg body weight, 7.0 mg / kg body weight, 7.1 mg / kg body weight, 7.2 mg / kg body weight, 7.3 mg / kg body weight, 7.4 mg / kg body weight, 7.5 mg / kg body weight, 7.6 mg / kg body weight, 7.7 mg / kg body weight, 7.8mg / kg body weight, 7.9mg / kg body weight, 8.0mg / kg body weight, 8.1mg / kg body weight, 8.2mg / kg body weight, 8.3mg / kg body weight, 8.4mg / kg body weight, 8.5mg / kg body weight, 8.6mg / kg body weight, 8.7mg / kg body weight, 8.8mg / kg body weight, 8.9mg / kg body weight, 9.0mg / kg body weight, 9.1mg / kg body weight, 9.2mg / kg body weight, 9.3mg / kg body weight, 9.4mg / kg body weight, 9.5mg / kg body weight, 9.6mg / kg body weight, 9.7mg / kg body weight, 9.8mg / kg body weight, 9.9mg / kg body weight, 10.0mg / kg Body weight, 10.1mg / kg body weight, 10.2mg / kg body weight, 10.3mg / kg body weight, 10.4mg / kg body weight, 10.5mg / kg body weight, 10.6mg / kg body weight, 10.7mg / kg body weight, 10.8mg / kg body weight, 10.9mg / kg body weight, 11.0mg / kg body weight, 11.1mg / kg body weight, 11.2mg / kg body weight, 11.3mg / kg body weight, 11.4mg / kg body weight, 11.5mg / kg body weight, 11.6mg / kg body weight, 11.7mg / kg body weight, 11.8mg / kg body weight, 11.9mg / kg body weight, 12.0mg / kg body weight, 12.1mg / kg bw, 12.2mg / kg bw, 12.3mg / kg bw, 12.4mg / kg bw, 12.5mg / kg bw, 12.6mg / kg bw, 12.7mg / kg bw, 12.8mg / kg bw, 12.9mg / kg bw, 13.0mg / kg bw, 13.1mg / kg bw, 13.2mg / kg bw, 13.3mg / kg bw, 13.4mg / kg bw, 13.5mg / kg bw, 13.6mg / kg bw, 13.7mg / kg bw, 13.8mg / kg bw, 13.9mg / kg bw, 14.0mg / kg bw, 14.1mg / kg bw, 14.2m g / kg body weight, 14.3mg / kg body weight, 14.4mg / kg body weight, 14.5mg / kg body weight, 14.6mg / kg body weight, 14.7mg / kg body weight, 14.8mg / kg body weight, 14.9mg / kg body weight, 15.0mg / kg body weight, 15.1mg / kg body weight, 15.2mg / kg body weight, 15.3mg / kg body weight, 15.4mg / kg body weight, 15.5mg / kg body weight, 15.6mg / kg body weight, 15.7mg / kg body weight, 15.8mg / kg body weight, 15.9mg / kg body weight, 16.0mg / kg body weight, 16.1mg / kg body weight, 16.2mg / kg body weight, 16.3 mg / kg body weight, 16.4 mg / kg or 16.5 mg / kg body weight or about 0.05-0.5 mg / kg body weight, 0.5-1.0 mg / kg body weight, 1.0-1.5 mg / kg body weight, 1.5-2.0 mg / kg body weight, 2.0-2.5 mg / kg body weight, 2.5-3.0 mg / kg body weight, 3.0-3.5 mg / kg body weight, 3.5-4.0 mg / kg body weight, 4.0-4.5 mg / kg body weight, 4.5-5.0 mg / kg body weight, 5.0-5.5 mg / kg body weight, 5.5-6.0 mg / kg body weight, 6.0-6.5 mg / kg body weight, 6.5-7.0 mg / kg body weight, 7.0-7.5 mg / kg body weight, 7.5-8.0 mg / kg body weight, 8.0-8 .5mg / kg body weight, 8.5-9.0mg / kg body weight, 9.0-9.5mg / kg body weight, 9.5-10.0mg / kg body weight, 10.0-10.5mg / kg body weight, 10.5-11.0mg / kg body weight, 11.0-11.5mg / kg body weight, 11.5-12.0mg / kg body weight, 12.0-12.5mg / kg body weight, 12.5-13.0mg / kg body weight, 13.0-13.5mg / kg body weight, 13.5-14.0mg / kg body weight, 14.0-14.5mg / kg body weight, 14.5-15.0mg / kg body weight, 15.0-15.5mg / kg body weight, 15.5-16.0mg / kg or 16.0-16.5mg / kg body weight. .

[0027] In some embodiments, the conjugate is administered at a dose that is the maximum tolerated dose (MTD).

[0028] In some embodiments, the conjugate is administered every day, every other day, every three days, every four days, every five days, 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.

[0029] In some embodiments, the conjugate is administered orally, intravenously, subcutaneously, intramuscularly, or topically.

[0030] In some embodiments, the methods of the invention do not cause unacceptable hyperglycemia in the subject.

[0031] In a specific embodiment, provided herein is a method for treating an autoimmune disease in a subject in need thereof by administering an effective amount of a conjugate, wherein the conjugate comprises 765IGF (SEQ ID NO: 2) and methotrexate.

[0032] In another specific embodiment, provided herein is a method for monitoring the effectiveness of a treatment of an autoimmune disease in a subject in need thereof using modulation of the frequency of cells expressing IGF-1R, the method comprising: administering to the subject an effective amount of a conjugate, wherein the conjugate comprises (i) an IGF-1R ligand or a portion or variant thereof, and (ii) a disease modifying agent; and subsequently determining the frequency of cells expressing IGF-1R in a sample obtained from the subject.

[0033] Other purposes and advantages will be explained in part in the following description, and in part will be clear from the description, or can be learned through practice. These purposes and advantages will be realized and obtained by the elements and combinations particularly pointed out in the appended claims.

[0034] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figures 1A-1E The effects of LX-101 on bulk subsets of peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and systemic sclerosis (SSc) are shown compared to vehicle control. Figure 1A )、CD4 + T cells ( Figure 1B )、CD8 + T cells ( Figure 1C ), NK cells( Figure 1D ) and NKT cells ( Figure 1E ) Subpopulations are shown relative to the percentage of viable cells determined by flow cytometry. Similar results were observed for vehicle control treated cells and LX-101 treated cells.

[0036] Figure 2A-2E Shown is the effect of LX-101 on IGF-1R in PBMCs from patients with systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and systemic sclerosis (SSc) compared to vehicle control. + The influence of subpopulations. Targeting B cells ( Figure 2A )、CD4 + T cells ( Figure 2B )、CD8 + T cells ( Figure 2C ), NK cells( Figure 2D ) and NKT cells ( Figure 2E ) subpopulations show relative activity of IGF-1R +Compared to cells treated with vehicle control, for cells treated with LX-101, an increase in the number of live IGF-1R + The percentage of cells decreased.

[0037] Figures 3A-3J Flow cytometry dot plots of PBMCs from SLE patients are shown. The dot plots show the expression levels of IGF-1R (CD221) in specific PBMC subsets. Figure 3A , 3C In 3, 3E, 3G and 3I, cells were treated with vehicle control. Figure 3B , 3D , 3F, 3H, and 3J, cells were treated with 17 μM LX-101. Figure 3A-3B CD4 + T cell populations. Figure 3C-3D CD8 + T cell populations. Figure 3E-3F CD19 + B cell populations. Figure 3G-3H CD56 + CD3 + NKT cell population. Figures 3I-3J CD56 + CD3 - NK cell population.

[0038] Figures 4A-4J Flow cytometry dot plots of PBMCs from RA patients are shown. The dot plots show the expression levels of IGF-1R (CD221) in specific PBMC subsets. Figure 4A , 4C In 4E, 4G and 4I, cells were treated with vehicle control. Figure 4B , 4D , 4F, 4H, and 4J, cells were treated with 17 μM LX-101. Figure 4A-4B CD4 + T cell populations. Figure 4C-4D CD8 + T cell populations. Figures 4E-4F CD19 + B cell populations. Figure 4G-4H CD56 + CD3 + NKT cell population. Figures 4I-4J CD56 + CD3 - NK cell population.

[0039] Figures 5A-5JFlow cytometry dot plots of PBMCs from SSc patients are shown. The dot plots show the expression levels of IGF-1R (CD221) in specific PBMC subsets. Figure 5A , 5C In 5E, 5G and 5I, cells were treated with vehicle control. Figure 5B , 5D , 5F, 5H, and 5J, cells were treated with 17 μM LX-101. Figure 5A-5B CD4 + T cell populations. Figure 5C-5D CD8 + T cell populations. Figure 5E-5F CD19 + B cell populations. Figure 5G-5H CD56 + CD3 + NKT cell population. Figures 5I-5J CD56 + CD3 - NK cell population. DETAILED DESCRIPTION

[0040] The currently disclosed subject matter will now be described more fully below. However, benefiting from the teaching presented in the foregoing description, the technical personnel in the field to which the currently disclosed subject matter is related will think of many modifications and other embodiments of the currently disclosed subject matter set forth herein. Therefore, it should be understood that the currently disclosed subject matter is not limited to the disclosed specific embodiments, and modifications and other embodiments are intended to be included within the scope of the appended claims. In other words, the subject matter described herein covers all alternatives, modifications and equivalent forms. If one or more of the combined documents, patents and similar materials are different from or contradictory to the present application, including but not limited to defined terms, term usage, described technology, etc., the present application shall prevail. Unless otherwise defined, all technical terms and scientific terms used herein have the same meaning as the meaning generally understood by those of ordinary skill in the art. All publications, patent applications, patents and other references mentioned herein are incorporated herein by reference in their entirety.

[0041] I. Definitions

[0042] As used herein, a patient or subject, etc. is any mammal suffering from an autoimmune disease. 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.

[0043] 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.

[0044] 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 causes cell killing can be used in the conjugates and methods of treating autoimmune diseases.

[0045] As used herein, the term "residue" or "residue of" a chemical moiety or compound refers to a chemical moiety or compound that is bound to a molecule whereby, by binding, at least one covalent bond has replaced at least one atom of the original chemical moiety or compound, thereby producing the residue of the chemical moiety or compound in the molecule.

[0046] As used herein, a subject is "refractory" to prior therapy if the subject fails to achieve a response to therapy such that the therapy is determined to be therapeutically ineffective (e.g., failure to achieve a clinical endpoint (including any response), prolonged duration of response, disease-free survival, relapse-free survival, and progression-free survival).

[0047] As used herein, "and / or" refers to and encompasses any and all possible combinations of one or more of the associated listed items as well as the lack of a combination when interpreted in the alternative ("or").

[0048] As used herein, when referring to a measurable value such as an amount of a compound or agent of the present subject matter, dosage, time, temperature, etc., the term "about" is meant to encompass variations of ±20%, ±10%, ±5%, ±1%, ±0.5%, or even ±0.1% of the specified amount.

[0049] As used herein, conditional language used herein, such as "may," "can," "might," "could," "for example," and the like, unless otherwise specifically stated or otherwise understood in the context of use, is generally intended to convey that certain embodiments include certain features, elements, and / or steps, while other embodiments do not include certain features, elements, and / or steps. Thus, such conditional language is generally not intended to imply in any way that features, elements, and / or steps are required for one or more embodiments, or that one or more embodiments must include logic for determining whether such features, elements, and / or steps are included in any particular embodiment or to be performed, with or without author input or prompting. The terms "comprising," "including," "having," and the like are synonymous and are used in an inclusive manner and do not exclude additional elements, features, actions, operations, and the like. Likewise, the term "or" is used in its inclusive sense (and not in its exclusive sense), so that, for example, when used to connect a list of elements, the term "or" means one, some, or all of the elements in the list.

[0050] Definitions of additional terms may be set forth below.

[0051] II. Composition

[0052] Drugs used for treatment

[0053] The present application provides a method for treating an autoimmune disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a conjugate, wherein the conjugate comprises an IGF-1R ligand or a portion or variant thereof and a disease-modifying agent.

[0054] In this or any embodiment of the invention, the conjugate may comprise a chemical conjugate in which the IGF-1R ligand and the disease modifying agent are chemically linked together directly or through a chemical linker. In other embodiments, the conjugate is a genetic 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 by a peptide bond.

[0055] In certain embodiments, the conjugate is a fusion protein described in U.S. Patent No. 9,675,671, which is hereby incorporated by reference in its entirety.

[0056] IGF-1R ligand

[0057] IGF-1R is a heterotetramer composed of two extracellular ligand-binding a subunits and two transmembrane b subunits with kinase activity, and the kinase activity mediates signal transduction. The natural ligands of IGF-1R are IGF-1, IGF-2 and insulin. IGF-1R has the highest affinity for IGF-1, followed by affinity for IGF-2, and can bind to insulin with an affinity of 1 / 100 to 1 / 50 of the affinity. IGF-1R can also form a mixed receptor by dimerization with the insulin receptor. See Hakuno et al. Journal of Molecular Endocrinology (J Mol Endocrinol.) 61 (1): T69-T86 (2018).

[0058] In certain embodiments, the IGF-1R ligand in the conjugates of the invention comprises wild-type IGF-1 (SEQ ID NO: 3), wild-type insulin (SEQ ID NO: 10 and SEQ ID NO: 11; mature insulin consists of two chains connected by disulfide bonds (chain A corresponding to SEQ ID NO: 10 and chain B corresponding to SEQ ID NO: 11), so two SEQ ID NOs are recited) or wild-type IGF-2 (SEQ ID NO: 12). In other embodiments, the IGF-1R ligand in the conjugates comprises a variant of wild-type IGF-1 (SEQ ID NO: 3), a variant of wild-type insulin (SEQ ID NO: 10 and SEQ ID NO: 11), or a variant of wild-type IGF-2 (SEQ ID NO: 12). In a specific embodiment, 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 wild-type insulin is at least 70%, 75%, 80%, 85%, 90%, 95% or 99% identical to insulin (SEQ ID NO:10 and SEQ ID NO:11), or the 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).

[0059] In another specific embodiment, the IGF-1R ligand in the conjugate comprises a variant of IGF-1 that has a reduced binding affinity to IGFBP compared to the binding affinity of wild-type IGF-1 (SEQ ID NO: 3) to the IGFBP or a variant of IGF-2 that has a reduced binding affinity to IGFBP compared to the binding affinity of wild-type IGF-2 (SEQ ID NO: 12). IGFBPs belong to a family of at least six proteins that bind to IGF-1 and IGF-2 with high affinity. IGFBPs bind to most IGFs in circulation, thereby increasing their half-life, regulating their bioavailability, and generally inhibiting their ability to bind to IGF receptors. See Baxter, 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 that have reduced binding to IGFBPs are more biologically active in vivo.

[0060] IGF-1 variants with reduced binding affinity for IGFBPs are known in the art and include: IGF132 (disclosed in U.S. Pat. No. 4,876,242), in which 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), in which the glutamic acid in 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), which lacks 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., Journal of Molecular Endocrinology 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).

[0061] In another embodiment, the IGF-1 variant has a higher affinity for IGF-1R than wild-type IGF-1 has for said IGF-1R, or the IGF-2 variant has a higher affinity for IGF-1R than wild-type IGF-2 has for said IGF-1R.

[0062] In another embodiment, the IGF-1R ligand or a portion or variant thereof comprises a leader sequence. The leader sequence may incorporate a tag (such as a polyhistidine tag) that is used to facilitate protein purification and provide a site for conjugation of a disease-modifying agent. In a specific embodiment, the leader sequence comprises SEQ ID NO: 1.

[0063] 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 facilitates protein purification and provides a site for conjugation of a disease-modifying agent as described above. 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 methionyl porcine 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 methionyl porcine 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 methionyl porcine growth hormone, followed by a Val-Asn dipeptide, followed by a variant of human IGF-1 in which the glutamic acid in position 3 of native human IGF-1 (SEQ ID NO:3) is substituted with glycine. In some embodiments, the conjugate comprises 765IGF (SEQ ID NO:2).

[0064] Disease Modifying Agents

[0065] As used herein, the term disease modifying agent refers to any agent that is capable of reducing and / or reversing the activity, severity and / or progression of a disease when treated according to the methods described herein.

[0066] In a specific embodiment, the disease modifying agent of the conjugate is a cytotoxic agent.Any suitable cytotoxic agent that causes cell killing may be used in the conjugates and methods of treating autoimmune diseases of the invention.

[0067] In a specific embodiment, the cytotoxic agent is a chemotherapeutic agent. Any suitable chemotherapeutic agent that causes cell killing can be used in the conjugates and methods of treating autoimmune diseases of the present invention. For example, in a specific embodiment, the chemotherapeutic agent is amsacrine, azacytidine, bleomycin, busulfan, capecitabine, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, cyclophosphamide, cytarabine, dactinomycin, daunorubicin, dacarbazine, docetaxel, doxorubicin, epirubicin, estramustine, etoposide, floxuridine, fludarabine, fluorouracil, gemcitabine, hexamethonium, idarubicin, isocyclobutanone ...aunorubicin, daunorubicin, daunorubicin, daunorubicin, daunorubicin, daunorubicin, daunorubicin, daunorubicin, Phosphamide, irinotecan, lomustine, nitrogen mustard, melphalan, mercaptopurine, methotrexate, mitomycin C, mitotane, mitoxantrone, oxaliplatin, paclitaxel, pemetrexed, pentostatin, plicamycin, procarbazine, raltitrexed, semustine, streptozotocin, temozolomide, teniposide, thioguanine, thiotepa, topotecan, trimetrexate, valrubicin, vincristine, vinblastine, vindesine or vinorelbine. In a specific embodiment, the chemotherapeutic agent is methotrexate.

[0068] In certain embodiments, the conjugate comprises more than one cytotoxic agent bound to an IGF-1R ligand. In certain aspects, the conjugate may comprise one to 12 cytotoxic agents, or 6 to 10 cytotoxic agents, or about 8 cytotoxic agents. In certain aspects, the conjugate may comprise one to twelve covalently bound cytotoxic agents, or 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 is covalently bound to any available lysine (when present) in the leader sequence.

[0069] In certain embodiments, the subject matter described herein relates to a method for treating an autoimmune disease 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, wherein the methotrexate is covalently bound to a lysine of SEQ ID NO:2, and the autoimmune disease is selected from the group consisting of rheumatoid arthritis, Graves' disease, SSc, Cushing's syndrome, idiopathic pulmonary fibrosis, SLE, and Crohn's disease. In one aspect of these embodiments, the conjugate is LX-101 (a conjugate as described above, wherein the IGF-1R ligand is SEQ ID NO:2, the cytotoxic agent is methotrexate, and wherein at least 6 and at most 10, or at least 6 and at most 9, or at least 7 and at most 9, or at least 8 and at most 9 methotrexate residues are each covalently bound to a lysine residue of SEQ ID NO:2).

[0070] 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.

[0071] In other embodiments, the cytotoxic agent is a toxin. Any suitable toxin that causes cell killing can be used in the conjugates of the present invention and methods for treating autoimmune diseases. The toxin can be derived from plants, fungi or bacteria. For example, in specific embodiments, the toxin is a Clostridium perfringens enterotoxin, a diphtheria toxin, a ricin A chain, a deglycosylated ricin A chain, a Pseudomonas exotoxin, an A chain toxin, a ribosome inactivating protein, an α-sarcosyl protein, a fusin, abrin, a restrictomycin, a bacterial endotoxin, a lipid A portion of a bacterial endotoxin, a cholera toxin or a ribonuclease. In a specific embodiment, the toxin comprises a portion of a Clostridium perfringens enterotoxin or a variant thereof. In a specific embodiment, the conjugate comprises SEQ ID NO: 14 or SEQ ID NO: 15. In other specific embodiments, the toxin comprises a diphtheria toxin or a portion or variant thereof. In a specific embodiment, the conjugate comprises SEQ ID NO: 13 or SEQ ID NO: 16.

[0072] Table 1 provides a listing of certain sequences cited herein.

[0073]

[0074]

[0075] Pharmaceutical composition

[0076] The conjugates of the invention can be formulated in pharmaceutical compositions for use in the methods described herein. In some embodiments, the pharmaceutical composition comprises an effective amount of the conjugates of the 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 allows the composition to be administered to a subject.

[0077] The material used to prepare the pharmaceutical composition can be non-toxic in the amount used. It is obvious to those of ordinary skill in the art that the optimal dose of the active ingredient in the pharmaceutical composition will depend on multiple factors. Related factors include, but are not limited to, the type of subject (e.g., human), the overall health level of the subject, the type of the condition suffered by the subject, the use of the composition as a part of a multi-drug regimen, the specific form of the composition of the present invention, and the mode of administration. The pharmaceutical composition comprises an effective amount of the composition of the present invention so that a suitable dosage will be obtained.

[0078] The term "carrier" refers to a diluent, adjuvant or excipient to be administered with the composition of the present invention. Any auxiliary agent, stabilizer, thickener, lubricant and colorant may be used. In one embodiment, the composition of the present invention and the pharmaceutically acceptable carrier are sterile when administered to a subject. When the composition of the present invention is administered intravenously, water may be a carrier. Saline solutions and aqueous solutions of dextrose and glycerol may also be used as liquid carriers, particularly for injectable solutions. If necessary, the composition of the present invention may also contain a small amount of pH buffer.

[0079] Liquid compositions of the present invention, whether they are solutions, suspensions or other similar forms, may also include one or more of the following: sterile diluents, such as water for injection, saline solution, physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils (such as synthetic monoglycerides or diglycerides, which can be used as solvents or suspension media), polyethylene glycol, glycerol, cyclodextrin, propylene glycol or other solvents; antibacterial agents, such as benzyl alcohol or methyl paraben; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers, such as acetates, citrates or phosphates; agents for adjusting pH, such as hydrochloric acid; and agents for adjusting tension, such as sodium chloride or dextrose. Parenteral compositions may be packaged in ampoules, disposable syringes or multiple dose vials made of glass, plastic or other materials. In some embodiments, physiological saline is an adjuvant. Injectable compositions may be sterile.

[0080] The compositions of the present invention can take the form of solutions, suspensions, tablets, pills, granules, capsules, capsules containing liquids, powders, sustained release formulations, suppositories, emulsions, aerosols, sprays, suspensions or any other form suitable for use. Examples of suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences by EW Martin.

[0081] In certain embodiments, the composition of the present invention is formulated as a pharmaceutical composition suitable for intravenous administration to human subjects according to conventional procedures. Generally, the carrier or vehicle for intravenous administration is a sterile isotonic aqueous buffer solution. If necessary, the composition may also include a solubilizing agent. The composition for intravenous administration may optionally include a local anesthetic, such as lidocaine (lignocaine) for relieving pain at the injection site. Generally, the components are separated or mixed together to provide in unit dosage form, for example, as a dry lyophilized powder or anhydrous concentrate in an airtight sealed container (such as an ampoule or a sachet indicating the amount of the active agent). When the composition of the present invention is to be administered by infusion, it can be, for example, distributed with an infusion bottle containing sterile pharmaceutical grade water or saline. When the composition of the present invention is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the components can be mixed before administration.

[0082] Pharmaceutical compositions can be prepared using methods well known in the pharmaceutical field. For example, compositions intended to be administered by injection can be prepared by combining the compositions of the present invention with water to form a solution. Surfactants can be added to promote the formation of homogeneous solutions or suspensions. Surfactants are complexes that can non-covalently interact with the compositions of the present invention to promote dissolution or homogeneous suspension of the compositions of the present invention in an aqueous delivery system.

[0083] III. Methods

[0084] Methods for producing IGF-1R conjugates

[0085] Methods for producing the conjugates of the invention are known in the art.

[0086] Nucleotide sequences encoding IGF-1R ligands can be produced 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 purified by any method known in the art for purifying proteins.

[0087] Conjugates of the invention comprising an IGF-IR ligand and a chemotherapeutic agent can be prepared by standard chemical and protein conjugation techniques and are described in US Pat. Nos. 7,811,982; 9,675,671; and 9,801,923.

[0088] Conjugates of the invention comprising an IGF-1R ligand and a toxin can be prepared as fusion proteins by standard recombinant DNA techniques and are described in US Pat. No. 8,017,102.

[0089] Treatment

[0090] Provided herein are methods of treating autoimmune diseases using conjugates, wherein the conjugates comprise an IGF-1R ligand, or a portion or variant thereof, and a disease modifying agent.

[0091] Non-limiting examples of autoimmune diseases that can be treated by the methods of the present invention include adrenergic drug resistance, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, adrenal autoimmune disease, allergic encephalomyelitis, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inflammatory eye disease, autoimmune neonatal thrombocytopenia, autoimmune neutropenia, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, autoimmune thyroiditis, Behcet's disease, bullous pemphigoid, myocardial infarction, and leukemia. disease, cardiotomy syndrome, celiac disease-dermatitis, chronic active hepatitis, chronic fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, Chargar-Strauss syndrome, cicatricial pemphigoid, CREST syndrome, cold agglutinin disease, Crohn's disease, Cushing's syndrome, cutaneous graft-versus-host disease (GVHD), dense deposit disease, dermatomyositis, essential mixed cryoglobulinemia, fibromyalgia-fibromyositis, glomerulonephritis (e.g., IgA nephropathy), gluten-sensitive enteropathy, Goodpasture's syndrome, Graves' disease, Guillain-Barré disease, hyperthyroidism (i.e., Hashimoto's thyroiditis), idiopathic pulmonary fibrosis, idiopathic Addison's disease, idiopathic thrombocytopenic purpura (ITP), IgA neuropathy, inflammatory arthritis, irritable bowel disease, juvenile arthritis, lichen planus, lichen sclerosus, lupus (e.g., SLE, cutaneous lupus, discoid lupus), Meniere's disease, mixed connective tissue disease, morphea, multiple sclerosis, myasthenia gravis, myocarditis, type 1 or immune-mediated diabetes mellitus, neuritis, other endocrine gland failure, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polymorrhage Osteitis, polyendocrinopathy, polyglandular syndromes, polymyalgia rheumatica, polymyositis, post-MI complications, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud's phenomenon, relapsing polychondritis, Reiter's syndrome, rheumatic heart disease, rheumatoid arthritis, sarcoidosis, Sjögren's syndrome, stiff-person syndrome, systemic sclerosis (SSc), Takayasu's arteritis, temporal arteritis / giant cell arteritis, ulcerative colitis, urticaria, uveitis, uveitic ophthalmia, vasculitis such as dermatitis herpetiformis, vitiligo, and Wegener's granulomatosis.

[0092] In a specific embodiment, the present disclosure provides a method of treating rheumatoid arthritis, Graves' disease, SSc, Cushing's syndrome, idiopathic pulmonary fibrosis, SLE, or Crohn's disease in a subject in need thereof.

[0093] In another specific embodiment, the present disclosure provides a method of treating diffuse cutaneous SSc in a subject in need thereof.

[0094] Methods for measuring clinical efficacy

[0095] As used herein, the terms "subject" and "patient" are used interchangeably.

[0096] In some embodiments, the method for treating an autoimmune disease in a subject in need thereof with a conjugate of the invention results in remission of the subject's disease. As used herein, disease remission may refer to any prevention, delay, reduction, and / or reversal of the activity, severity, and / or progression of a subject's disease when treated according to the methods of the invention. Disease remission may be caused by a direct or indirect effect on the underlying pathophysiology of the disease due to treatment with a conjugate of the invention.

[0097] The method of evaluating autoimmune patients to determine the improvement of disease symptoms or the reduction of disease is known to those skilled in the art. For example, the improvement of disease symptoms or the reduction of disease can be assessed by disease activity score or index [e.g., disease activity score 28 (DAS28) of rheumatoid arthritis; SLE disease activity index (SLEDAI) of SLE]. See van Riel et al., "Clin Exp Rheumatol", 34 (5th Supplement 101): S40-S44 (2016) and Ohmura, "Modern Rheumatology (Mod Rheumatol)", 31 (1): 20-28 (2021). The effect of therapy can be monitored by continuously assessing the disease activity score by comparing the score at a later time point with the score at a previous time point (e.g., before administering therapy) throughout the treatment process. Biomarkers predicting clinical benefit (e.g., rheumatoid factor and autoantibodies to citrullinated proteins in rheumatoid arthritis; antinuclear antibodies and serum complement levels in SLE) can also be used to assess disease reduction, and the biomarkers are known in the art. See Shapiro, Cureus. 13(5):e15063 (2021) and Yu et al., Biomolecules. 11(7):928 (2021).

[0098] In some embodiments, the method for treating an autoimmune disease in a subject in need thereof with a conjugate of the invention results in functional alteration or impairment of IGF-1R-expressing cells in the subject. As used herein, alteration or impairment refers to any change in the endogenous function of the IGF-1R-expressing cells in an autoimmune disease.

[0099] In some embodiments, the method for treating an autoimmune disease in a subject in need thereof with a conjugate of the invention results in a reduction in the number of cells expressing IGF-1R in the subject. In a specific embodiment, the reduction is caused by killing of cells expressing IGF-1R. Without being bound by theory, it is believed that simple inhibition of the IGF-1R pathway may be insufficient as a completely curative treatment due to signaling through other potentially redundant pathways, or compensatory mechanisms, or pathways that are upregulated and may lead to adaptive resistance that may continue to act as disease effectors. Therefore, targeted delivery of disease-modifying agents (including agents that result in killing of cells expressing IGF-1R) can provide benefits beyond (including superior to) simple IGF-1R pathway inhibition because it bypasses the effects of all such potentially redundant and / or compensatory pathways.

[0100] In a specific embodiment, the cells expressing IGF-1R are B lymphocytes and / or T lymphocytes. Autoreactive B lymphocytes can produce pathogenic autoantibodies that can cause tissue damage and / or dysregulate cell signaling, while autoreactive T lymphocytes can activate autoreactive B cells and lead to inflammation and tissue damage.

[0101] In some embodiments, IGF-1R is overexpressed by cells in subjects diagnosed with an autoimmune disease relative to expression by cells from healthy subjects or subjects not diagnosed with an autoimmune disease. In some embodiments, the frequency of cells expressing IGF-1R is increased in subjects diagnosed with an autoimmune disease relative to the frequency of cells from healthy subjects or subjects not diagnosed with an autoimmune disease. In some embodiments, the frequency of cells expressing IGF-1R is measured by flow cytometry or immunohistochemistry using techniques and cell surface markers known in the art.

[0102] In certain embodiments, the reduction in the number of cells expressing IGF-1R obtained by the methods of the present invention results in an improvement in disease symptoms or a reduction in disease. Thus, also provided herein is a method for monitoring the effectiveness of treatment of an autoimmune disease in a subject in need thereof using modulation of the frequency of cells expressing IGF-1R, the method comprising: administering to the subject an effective amount of a conjugate, wherein the conjugate comprises (i) an IGF-1R ligand or a portion or variant thereof, and (ii) a disease modifying agent; and subsequently determining the frequency of cells expressing IGF-1R in a sample obtained from the subject. For example, treatment may reduce the frequency of IGF-1R expressing cells in a subject by at least 5%, 10%, 15%, 20%, 25%, 30%, 33%, 35%, 40%, 45%, 50%, 55%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more compared to the frequency of IGF-1R expressing cells in the subject before undergoing treatment with the conjugate of the invention. In certain embodiments, the frequency of IGF-1R expressing cells in a subject is reduced by about 20% to about 95% compared to the level before treatment began. For example, in certain embodiments, the frequency of IGF-1R expressing cells in a subject is reduced by about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80% or about 90% compared to the level before treatment began.

[0103] In certain embodiments, the sample used to determine the frequency of cells expressing IGF-1R is a peripheral blood mononuclear cell (PBMC) sample. Methods of PBMC isolation are well known in the art.

[0104] Combination therapy

[0105] In some embodiments, the conjugates of the present invention are administered in combination with one or more other therapies. Any therapy (e.g., therapeutic agent or preventive agent) approved, available, used, or currently used for the treatment and / or management of autoimmune diseases by the U.S. Food and Drug Administration (FDA) can be used in combination with the conjugates of the present invention, including: prednisone, hydroxychloroquine, chloroquine, belimumab, anilumab, abatacept, atacicept, lupuzor, rituximab, cyclosporine, aldesleukin, baricitinib, BIIB059, BI655064, bortezomib, BT063, selimud, pegol dapilozumab, efretide, filgotinib, GS-9876, iberdomide, IFN-α kinin, elamod, nelfinavir, obotuzumab, OMS721, rapamycin, RC18, RSL V-132, SM101, cilalizumab, ustekinumab, wolbalizumab, XmAb5871, bilimud, tabecirumab, epratuzumab, regremod, tacrolimus, longizumab, sifalimumab, aniluzumab, tocilizumab, infliximab, metilimumab, fesutuzumab, rilonacept, cyclophosphamide, methotrexate, nintedanib, JBT-101, imatinib, pirfenidone, nilotinib, dasatinib, SAR100842, BMS-986202, BAY41-2272, riociguat, risunab, ixekizumab, berdalumab, troloxetine, etanercept, adalimumab, golimumab, secukinumab, teikizumab, tofacitinib, and guselkumab.

[0106] In some embodiments, the combinations of agents disclosed herein allow for administration of one or more agents at dosage levels lower than the dosage at which the agents would be effective when administered as single agents.

[0107] Dosing and administration

[0108] The agents of the present invention may be administered at any clinically relevant dose. Clinically relevant means that the dose of the agent has an effect on the subject.

[0109] In some embodiments, the conjugate of the invention is administered at about 0.05 μEq / kg body weight, 0.10 μEq / kg body weight, 0.20 μEq / kg body weight, 0.40 μEq / kg body weight, 0.80 μEq / kg body weight, 1.0 μEq / kg body weight, 1.5 μEq / kg body weight, 1.6 μEq / kg body weight, 2.0 μEq / kg body weight, 2.5 μEq / kg body weight, 3.0 μEq / kg body weight, 3.5 μEq / kg body weight, 4.0 μEq / kg body weight g body weight, 4.5 μEq / kg body weight, 5.0 μEq / kg body weight, 5.5 μEq / kg body weight, 6.0 μEq / kg body weight, 6.5 μEq / kg body weight, 7.0 μEq / kg body weight, 7.5 μEq / kg body weight, 8.0 μEq / kg body weight, 8.5 μEq / kg body weight, 9.0 μEq / kg body weight or 10.0 μEq / kg body weight, where μEq is equivalent to mmol of the chemotherapeutic agent conjugated to the IGF-1R ligand.

[0110] In some embodiments, the conjugate of the invention is administered at about 0.05-0.5 μEq / kg body weight, 0.5-1.0 μEq / kg body weight, 1.0-1.5 μEq / kg body weight, 1.5-2.0 μEq / kg body weight, 2.0-2.5 μEq / kg body weight, 2.5-3.0 μEq / kg body weight, 3.0-3.5 μEq / kg body weight, 3.5-4.0 μEq / kg body weight, 4.0-4.5 μEq / kg body weight, 4.5-5.0 μEq / kg body weight, 5.0-5.5 μEq / kg body weight. kg body weight, 5.5-6.0 μEq / kg body weight, 6.0-6.5 μEq / kg body weight, 6.5-7.0 μEq / kg body weight, 7.0-7.5 μEq / kg body weight, 7.5-8.0 μEq / kg body weight, 8.0-8.5 μEq / kg body weight, 8.5-9.0 μEq / kg body weight, 9.0-9.5 μEq / kg body weight or 9.5-10.0 μEq / kg body weight, where μEq is equivalent to mmol of the chemotherapeutic agent conjugated to the IGF-1R ligand.

[0111] In some embodiments, the conjugate of the invention is administered at about 0.05 mg / kg body weight, 0.10 mg / kg body weight, 0.15 mg / kg body weight, 0.20 mg / kg body weight, 0.25 mg / kg body weight, 0.30 mg / kg body weight, 0.35 mg / kg body weight, 0.40 mg / kg body weight, 0.45 mg / kg body weight, 0.50 mg / kg body weight, 0.55 mg / kg body weight, 0.60 mg / kg body weight, 0.65 mg / kg body weight, 0.70 mg / kg body weight, 0.75 mg / kg body weight, 0.80 mg / kg body weight, 0.85 mg / kg body weight, 0.90 mg / kg body weight, 0.95 mg / kg body weight, 1.0 mg / kg body weight, 1.1 mg / kg body weight, 1.2 mg / kg body weight, 1.3 mg / kg body weight, 1.4 mg / kg body weight, 1.5 mg / kg body weight, 1.6 mg / kg body weight, 1.7 mg / kg body weight, 1.8 mg / kg body weight, 1.9 mg / kg body weight, 2.0 mg / kg body weight, 2.1 mg / kg body weight, 2.2 mg / kg body weight, 2.3 mg / kg body weight, 2.4 mg / kg body weight, 2.5 mg / kg body weight, 2.6 mg / kg body weight, 2.7 mg / kg body weight, 2.8 mg / kg body weight, 2.9 mg / kg body weight, 3.0 mg / kg body weight, 3.1 mg / kg body weight, 3.2 mg / kg body weight , 3.3 mg / kg body weight, 3.4 mg / kg body weight, 3.5 mg / kg body weight, 3.6 mg / kg body weight, 3.7 mg / kg body weight, 3.8 mg / kg body weight, 3.9 mg / kg body weight, 4.0 mg / kg body weight, 4.1 mg / kg body weight, 4.2 mg / kg body weight, 4.3 mg / kg body weight, 4. 4mg / kg body weight, 4.5mg / kg body weight, 4.6mg / kg body weight, 4.7mg / kg body weight, 4.8mg / kg body weight, 4.9mg / kg body weight, 5.0mg / kg body weight, 5.1mg / kg body weight, 5.2mg / kg body weight, 5.3mg / kg body weight, 5.4mg / kg body weight, 5.5mg / kg body weight, 5.6 mg / kg body weight, 5.7 mg / kg body weight, 5.8 mg / kg body weight, 5.9 mg / kg body weight, 6.0 mg / kg body weight, 6.1 mg / kg body weight, 6.2 mg / kg body weight, 6.3 mg / kg body weight, 6.4 mg / kg body weight, 6.5 mg / kg body weight, 6.6 mg / kg body weight Weight, 6.7 mg / kg body weight, 6.8 mg / kg body weight, 6.9 mg / kg body weight, 7.0 mg / kg body weight, 7.1 mg / kg body weight, 7.2 mg / kg body weight, 7.3 mg / kg body weight, 7.4 mg / kg body weight, 7.5 mg / kg body weight, 7.6 mg / kg body weight, 7.7 mg / kg body weight, 7.8mg / kg body weight, 7.9mg / kg body weight, 8.0mg / kg body weight, 8.1mg / kg body weight, 8.2mg / kg body weight, 8.3mg / kg body weight, 8.4mg / kg body weight, 8.5mg / kg body weight, 8.6mg / kg body weight, 8.7mg / kg body weight, 8.8mg / kg body weight, 8.9mg / kg body weight, 9.0mg / kg body weight, 9.1mg / kg body weight, 9.2mg / kg body weight, 9.3mg / kg body weight, 9.4mg / kg body weight, 9.5mg / kg body weight, 9.6mg / kg body weight, 9.7mg / kg body weight, 9.8mg / kg body weight, 9.9mg / kg body weight, 10.0mg / kg Body weight, 10.1mg / kg body weight, 10.2mg / kg body weight, 10.3mg / kg body weight, 10.4mg / kg body weight, 10.5mg / kg body weight, 10.6mg / kg body weight, 10.7mg / kg body weight, 10.8mg / kg body weight, 10.9mg / kg body weight, 11.0mg / kg body weight, 11.1mg / kg body weight, 11.2mg / kg body weight, 11.3mg / kg body weight, 11.4mg / kg body weight, 11.5mg / kg body weight, 11.6mg / kg body weight, 11.7mg / kg body weight, 11.8mg / kg body weight, 11.9mg / kg body weight, 12.0mg / kg body weight, 12.1mg / kg bw, 12.2mg / kg bw, 12.3mg / kg bw, 12.4mg / kg bw, 12.5mg / kg bw, 12.6mg / kg bw, 12.7mg / kg bw, 12.8mg / kg bw, 12.9mg / kg bw, 13.0mg / kg bw, 13.1mg / kg bw, 13.2mg / kg bw, 13.3mg / kg bw, 13.4mg / kg bw, 13.5mg / kg bw, 13.6mg / kg bw, 13.7mg / kg bw, 13.8mg / kg bw, 13.9mg / kg bw, 14.0mg / kg bw, 14.1mg / kg bw, 14.2m g / kg body weight, 14.3mg / kg body weight, 14.4mg / kg body weight, 14.5mg / kg body weight, 14.6mg / kg body weight, 14.7mg / kg body weight, 14.8mg / kg body weight, 14.9mg / kg body weight, 15.0mg / kg body weight, 15.1mg / kg body weight, 15.2mg / kg body weight, 15.3mg / kg body weight, 15.4mg / kg body weight, 15.5mg / kg body weight, 15.6mg / kg body weight, 15.7mg / kg body weight, 15.8mg / kg body weight, 15.9mg / kg body weight, 16.0mg / kg body weight, 16.1mg / kg body weight, 16.2mg / kg body weight, 16.3 mg / kg body weight, 16.4 mg / kg or 16.5 mg / kg body weight, where mg refers to the amount of IGF-1R ligand present in the conjugate.

[0112] In some embodiments, the conjugate of the invention is administered at about 0.05-0.5 mg / kg body weight, 0.5-1.0 mg / kg body weight, 1.0-1.5 mg / kg body weight, 1.5-2.0 mg / kg body weight, 2.0-2.5 mg / kg body weight, 2.5-3.0 mg / kg body weight, 3.0-3.5 mg / kg body weight, 3.5-4.0 mg / kg body weight, 4.0-4.5 mg / kg body weight, 4.5-5.0 mg / kg body weight, 5.0-5.5 mg / kg body weight, 5.5-6.0 mg / kg body weight, 6.0-6.5 mg / kg body weight, 6.5-7.0 mg / kg body weight, 7.0-7.5 mg / kg body weight, 7.5-8.0 mg / kg body weight, 8.0-8.5 mg / kg body weight, 8.5-9.0 mg / kg

[00136] In some embodiments, the conjugate is administered in a dosage range of 9.0-9.5 mg / kg body weight, 9.5-10.0 mg / kg body weight, 10.0-10.5 mg / kg body weight, 10.5-11.0 mg / kg body weight, 11.0-11.5 mg / kg body weight, 11.5-12.0 mg / kg body weight, 12.0-12.5 mg / kg body weight, 12.5-13.0 mg / kg body weight, 13.0-13.5 mg / kg body weight, 13.5-14.0 mg / kg body weight, 14.0-14.5 mg / kg body weight, 14.5-15.0 mg / kg body weight, 15.0-15.5 mg / kg body weight, 15.5-16.0 mg / kg or 16.0-16.5 mg / kg body weight, wherein mg refers to the amount of IGF-1R ligand present in the conjugate.

[0113] In certain embodiments, the conjugate of the present invention is administered with a maximum tolerated dose (MTD). As used herein, "MTD" refers to the highest dose of a medicament that a single patient can tolerate. In other words, the side effects of a given patient can determine the MTD. Side effects may limit the ability to administer a therapeutic dose higher than the maximum tolerated dose. Therefore, the MTD of a given patient may be lower than the dose indicated in the prescription information for treatment or the dose commonly used in clinical practice. MTD may have limited clinical efficacy or no clinical efficacy in patients.

[0114] In some embodiments, the conjugate of the invention is administered every day, every other day, every three days, every four days, every five days, 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.

[0115] The conjugates of the invention may be administered by any convenient route, such as by infusion or bolus injection, absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.). Administration may be systemic or local. Various delivery systems are known, such as microparticles, microcapsules, capsules, etc., and may be used to administer the compositions of the invention. Methods of administration may include, but are not limited to, oral administration and parenteral administration; parenteral administration, including but not limited to intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous; intranasal, epidural, sublingual, intracerebral, intraarterial, intraventricular, intrathecal, intravaginal, intracapsular, intraorbital, intracardiac, intradermal, transtracheal, subcutaneous, subcapsular, subarachnoid, intraspinal, intracerebrospinal, intrasternal, transdermal, rectal, vaginal, by inhalation, or topically to the ear, nose, eye, or skin.

[0116] The mode of administration is at the discretion of the practitioner and will depend in part on the site of the medical condition.

[0117] In some embodiments, the compositions of the invention are administered parenterally. In some embodiments, the compositions of the invention are administered intravenously. In another embodiment, the compositions of the invention are administered by continuous infusion. In a specific embodiment, the compositions of the invention are administered by infusion for 15 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, or 2 hours.

[0118] In some embodiments, it may be desirable to apply one or more compositions of the invention topically to an area in need of treatment. This may be achieved, for example, but not limited to, by local infusion during surgery; topical application, such as in conjunction with a wound dressing after surgery; by injection; by means of a catheter; by means of a suppository; or by means of an implant, which is a porous material, a non-porous material, or a colloidal material, including a membrane or fiber such as a silicone rubber membrane. In certain embodiments, one or more compositions of the invention may be injected intraperitoneally.

[0119] In yet another embodiment, the compositions of the present invention may be delivered in a controlled release system.

[0120] 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 can be, but is not limited to, an insulin-like pump.

[0121] In some embodiments, the conjugates of the invention are administered orally, intravenously, subcutaneously, intramuscularly, or topically.

[0122] In some embodiments, the methods of the invention do not cause unacceptable hyperglycemia in the subject. Hyperglycemia is another term for high blood glucose and may occur when there is not enough insulin in the body or the body cannot use insulin properly. Unacceptable hyperglycemia refers to a Grade 3 or higher side effect determined by the treating physician, and / or a side effect that cannot be controlled with diabetes medications and leads to termination of treatment with the conjugates of the invention.

[0123] Patient population

[0124] In some embodiments, the subject with the autoimmune disease treated according to the methods described herein has not previously received treatment for the autoimmune disease. In some embodiments, the subject with the autoimmune disease treated according to the methods described herein has previously received treatment for the autoimmune disease.

[0125] In some embodiments, the subject is refractory to a prior treatment for an autoimmune disease. As used herein, a subject is "refractory" to a prior treatment if the subject fails to achieve a response to a therapy such that the therapy is determined to be therapeutically ineffective (e.g., failure to achieve a clinical endpoint (including any response), prolonged duration of response, disease-free survival, relapse-free survival, progression-free survival, and prolonged overall survival).

[0126] The subject matter described herein includes, but is not limited to, the following examples:

[0127] 1. A method for treating an autoimmune disease in a subject, the method comprising administering to the subject an effective amount of a conjugate, wherein the conjugate comprises (i) an insulin-like growth factor 1 receptor (IGF-1R) ligand or a portion or variant thereof, and (ii) a disease-modifying agent.

[0128] 2. The method according to embodiment 1, wherein the autoimmune disease is selected from the group consisting of: adrenergic drug resistance, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, adrenal autoimmune disease, allergic encephalomyelitis, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inflammatory eye disease, autoimmune neonatal thrombocytopenia, autoimmune neutrophilia, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, autoimmune thyroiditis, Behcet's disease, bullous pemphigoid, cardiomyopathy, heart disease, incision syndrome, celiac disease-dermatitis, chronic active hepatitis, chronic fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, Chargar-Strauss syndrome, cicatricial pemphigoid, CREST syndrome, cold agglutinin disease, Crohn's disease, Cushing's syndrome, cutaneous graft-versus-host disease (GVHD), dense deposit disease, dermatomyositis, essential mixed cryoglobulinemia, fibromyalgia-fibromyositis, glomerulonephritis (eg, IgA nephropathy), gluten-sensitive enteropathy, Goodpasture's syndrome, Graves' disease, Guillain-Barré disease , hyperthyroidism (ie, Hashimoto's thyroiditis), idiopathic pulmonary fibrosis, idiopathic Addison's disease, idiopathic thrombocytopenic purpura (ITP), IgA neuropathy, inflammatory arthritis, irritable bowel disease, juvenile arthritis, lichen planus, lichen sclerosus, lupus [eg, systemic lupus erythematosus (SLE), cutaneous lupus, discoid lupus), Meniere's disease, mixed connective tissue disease, morphea, multiple sclerosis, myasthenia gravis, myocarditis, type 1 or immune-mediated diabetes mellitus, neuritis, other endocrine gland failure, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, Polychondritis, polyendocrinopathy, polyglandular syndrome, polymyalgia rheumatica, polymyositis, post-MI complications, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud's phenomenon, relapsing polychondritis, Reiter's syndrome, rheumatic heart disease, rheumatoid arthritis, sarcoidosis, Sjögren's syndrome, stiff-person syndrome, systemic sclerosis (SSc), Takayasu's arteritis, temporal arteritis / giant cell arteritis, ulcerative colitis, urticaria, uveitis, uveitic ophthalmia, vasculitis such as dermatitis herpetiformis angiitis, vitiligo, or Wegener's granulomatosis.

[0129] 3. The method of embodiment 2, wherein the autoimmune disease is rheumatoid arthritis, Graves' disease, SSc, Cushing's syndrome, idiopathic pulmonary fibrosis, SLE, or Crohn's disease.

[0130] 4. The method of embodiment 3, wherein the SSc is diffuse cutaneous SSc.

[0131] 5. The method of any one of embodiments 1 to 4, wherein said treatment results in functional alteration or damage of cells expressing IGF-1R in said subject.

[0132] 6. The method of any one of embodiments 1 to 4, wherein said treatment results in a decrease in the number of cells expressing IGF-1R in said subject.

[0133] 7. The method of claim embodiment 6, wherein said reduction is caused by killing of said cells expressing IGF-1R.

[0134] 8. The method of any one of embodiments 5 to 7, wherein the cells expressing IGF-1R are B lymphocytes and / or T lymphocytes.

[0135] 9. The method of any one of embodiments 1 to 8, wherein said IGF-1R is overexpressed by cells in said subject relative to expression by cells from a healthy subject or a subject not diagnosed as having said autoimmune disease.

[0136] 10. The method of any one of embodiments 1 to 9, wherein the frequency of cells expressing IGF-1R in the subject is increased relative to the frequency of cells from a healthy subject or a subject not diagnosed with the autoimmune disease.

[0137] 11. The method of embodiment 10, wherein the frequency of cells expressing IGF-1R is measured by flow cytometry or immunohistochemistry.

[0138] 12. The method of any one of embodiments 1 to 11, wherein said treatment results in disease remission in said subject.

[0139] 13. The method of any one of embodiments 1 to 12, wherein the IGF-1R ligand comprises wild-type insulin-like growth factor 1 (IGF-1) (SEQ ID NO:3), wild-type insulin (SEQ ID NO:10 and SEQ ID NO:11), or wild-type insulin-like growth factor 2 (IGF-2) (SEQ ID NO:12).

[0140] 14. The method of any one of embodiments 1 to 12, 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 NO:10 and SEQ ID NO:11), or a variant of wild-type IGF-2 (SEQ ID NO:12).

[0141] 15. A method according to embodiment 14, 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 NO:10 and SEQ ID NO:11), or the variant of wild-type IGF-2 is at least 90% identical to IGF-2 (SEQ ID NO:12).

[0142] 16. A method according to embodiment 14 or 15, wherein the binding affinity of the IGF-1 variant to insulin-like growth factor binding protein (IGFBP) is reduced compared to the binding affinity of wild-type IGF-1 (SEQ ID NO: 3) to the IGFBP, or the binding affinity of the IGF-2 variant to IGFBP is reduced compared to the binding affinity of wild-type IGF-2 (SEQ ID NO: 12) to the IGFBP.

[0143] 17. The method of any one of embodiments 14 to 16, wherein the IGF-1 variant has an increased affinity for the IGF-1R compared to the affinity of wild-type IGF-1 (SEQ ID NO:3) for the IGF-1R, or the IGF-2 variant has an increased affinity for the IGF-1R compared to the affinity of wild-type IGF-2 (SEQ ID NO:12) for the IGF-1R.

[0144] 18. The method of any one of embodiments 1 to 12 or embodiments 14 to 17, 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).

[0145] 19. The method of any one of embodiments 1 to 18, wherein the IGF-1R ligand, or a portion or variant thereof, comprises a leader sequence.

[0146] 20. The method of embodiment 19, wherein the leader sequence comprises SEQ ID NO: 1.

[0147] 21. The method of any one of embodiments 18 to 20, wherein said IGF-1R ligand comprises 765IGF (SEQ ID NO: 2).

[0148] 22. The method of any one of embodiments 1 to 21, wherein the disease-modifying agent comprises a cytotoxic agent.

[0149] 23. The method of embodiment 22, wherein the cytotoxic agent comprises a chemotherapeutic agent.

[0150] 24. The method of embodiment 23, wherein the chemotherapeutic agent is amsacrine, azacytidine, bleomycin, busulfan, capecitabine, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, cyclophosphamide, cytarabine, dactinomycin, daunorubicin, dacarbazine, docetaxel, doxorubicin, epirubicin, estramustine, etoposide, floxuridine, fludarabine, fluorouracil, gemcitabine, hexamethonium, idarubicin, or idarubicin. amphetamine, ifosfamide, irinotecan, lomustine, nitrogen mustard, melphalan, mercaptopurine, methotrexate, mitomycin C, mitotane, mitoxantrone, oxaliplatin, paclitaxel, pemetrexed, pentostatin, plicamycin, procarbazine, raltitrexed, semustine, streptozotocin, temozolomide, teniposide, thioguanine, thiotepa, topotecan, trimetrexate, valrubicin, vincristine, vinblastine, vindesine, or vinorelbine.

[0151] 25. The method of embodiment 24, wherein the chemotherapeutic agent is methotrexate.

[0152] 26. The method of embodiment 22, wherein the cytotoxic agent comprises a toxin.

[0153] 27. The method of embodiment 26, wherein the toxin comprises Clostridium perfringens enterotoxin, diphtheria toxin, ricin chain A, Pseudomonas exotoxin, A chain toxin, ribosome inactivating protein, alpha-sarcin, aspergillus, or ribonuclease.

[0154] 28. The method of embodiment 27, wherein the toxin comprises Clostridium perfringens enterotoxin or a portion or variant thereof.

[0155] 29. The method of embodiment 28, wherein the conjugate comprises SEQ ID NO:14 or SEQ ID NO:15.

[0156] 30. The method of embodiment 27, wherein the toxin comprises diphtheria toxin or a portion or variant thereof.

[0157] 31. The method of embodiment 30, wherein the conjugate comprises SEQ ID NO:13 or SEQ ID NO:16.

[0158] 32. A method according to any one of embodiments 1 to 31, wherein the subject (i) has not previously received treatment for the autoimmune disease; (ii) has previously received treatment for the autoimmune disease; (iii) has relapsed from a previous treatment for the autoimmune disease; or (iv) is refractory to a previous treatment for the autoimmune disease.

[0159] 33. The method of any one of embodiments 1 to 32, wherein the conjugate is administered in combination with one or more other therapies.

[0160] 34. The method of embodiment 33, wherein the one or more additional therapies comprise one or more of: prednisone, hydroxychloroquine, chloroquine, belimumab, anirumab, abatacept, atacicept, lupuzor, rituximab, cyclosporine, aldesleukin, baricitinib, BIIB059, BI655064, bortezomib, BT063, selimud, pegol dapilozumab, efretide, filgotinib, GS-9876, iberdocin, IFN-α kinin, iguratimod, nelfinavir, obinutuzumab, OMS721, rapamycin, RC18, RSLV-132, SM101, cilalizumab, ustekinumab Anti-, wolbalizumab, XmAb5871, bilimud, tabecirumab, epratuzumab, regremod, tacrolimus, longizumab, sifalimumab, anilumab, tocilizumab, infliximab, metilimab, fesutuzumab, rilonacept, cyclophosphamide, methotrexate, nintedanib, JBT-101, imatinib, pirfenidone, nilotinib, dasatinib, SAR100842, BMS-986202, BAY41-2272, riociguat, risunab, ixekizumab, berdalumab, trorocirumab, etanercept, adalimumab, golimumab, secukinumab, teikizumab, tofacitinib, and guselkumab.

[0161] 35. The method of any one of embodiments 1 to 25, wherein the conjugate is administered at about 0.05 μEq / kg body weight, 0.10 μEq / kg body weight, 0.20 μEq / kg body weight, 0.40 μEq / kg body weight, 0.80 μEq / kg body weight, 1.0 μEq / kg body weight, 1.5 μEq / kg body weight, 1.6 μEq / kg body weight, 2.0 μEq / kg body weight, 2.5 μEq / kg body weight, 3.0 μEq / kg body weight , 3.5 μEq / kg body weight, 4.0 μEq / kg body weight, 4.5 μEq / kg body weight, 5.0 μEq / kg body weight, 5.5 μEq / kg body weight, 6.0 μEq / kg body weight, 6.5 μEq / kg body weight, 7.0 μEq / kg body weight, 7.5 μEq / kg body weight, 8.0 μEq / kg body weight, 8.5 μEq / kg body weight, 9.0 μEq / kg body weight, 9.5 μEq / kg body weight or 10.0 μEq / kg body weight A heavy dose or about 0.05-0.5 μEq / kg body weight, 0.5-1.0 μEq / kg body weight, 1.0-1.5 μEq / kg body weight, 1.5-2.0 μEq / kg body weight, 2.0-2.5 μEq / kg body weight, 2.5-3.0 μEq / kg body weight, 3.0-3.5 μEq / kg body weight, 3.5-4.0 μEq / kg body weight, 4.0-4.5 μEq / kg body weight, 4.5-5.0 μEq / kg body weight, A dose range of 5.0-5.5 μEq / kg body weight, 5.5-6.0 μEq / kg body weight, 6.0-6.5 μEq / kg body weight, 6.5-7.0 μEq / kg body weight, 7.0-7.5 μEq / kg body weight, 7.5-8.0 μEq / kg body weight, 8.0-8.5 μEq / kg body weight, 8.5-9.0 μEq / kg body weight, 9.0-9.5 μEq / kg body weight or 9.5-10.0 μEq / kg body weight is administered.

[0162] 36. The method of any one of embodiments 1 to 34, wherein the conjugate is administered at about 0.05 mg / kg body weight, 0.10 mg / kg body weight, 0.15 mg / kg body weight, 0.20 mg / kg body weight, 0.25 mg / kg body weight, 0.30 mg / kg body weight, 0.35 mg / kg body weight, 0.40 mg / kg body weight, 0.45 mg / kg body weight, 0.50 mg / kg body weight, 0.55 mg / kg body weight, 0.60 mg / kg body weight, 0.65 mg / kg body weight, 0.70 mg / kg body weight, 0.75 mg / kg body weight, 0.80 mg / kg body weight, 0.85 mg / kg body weight, 0.90 mg / kg body weight, 0. 95 mg / kg body weight, 1.0 mg / kg body weight, 1.1 mg / kg body weight, 1.2 mg / kg body weight, 1.3 mg / kg body weight, 1.4 mg / kg body weight, 1.5 mg / kg body weight, 1.6 mg / kg body weight, 1.7 mg / kg body weight, 1.8 mg / kg body weight, 1.9 mg / kg body weight, 2.0 mg / kg body weight, 2.1 mg / kg body weight, 2.2 mg / kg body weight, 2.3 mg / kg body weight, 2.4 mg / kg body weight, 2.5 mg / kg body weight, 2.6 mg / kg body weight, 2.7 mg / kg body weight, 2.8 mg / kg body weight, 2.9 mg / kg body weight, 3.0 mg / kg body weight, 3.1 mg / kg body weight Weight, 3.2 mg / kg body weight, 3.3 mg / kg body weight, 3.4 mg / kg body weight, 3.5 mg / kg body weight, 3.6 mg / kg body weight, 3.7 mg / kg body weight, 3.8 mg / kg body weight, 3.9 mg / kg body weight, 4.0 mg / kg body weight, 4.1 mg / kg body weight, 4.2 mg / kg body weight, 4 .3mg / kg body weight, 4.4mg / kg body weight, 4.5mg / kg body weight, 4.6mg / kg body weight, 4.7mg / kg body weight, 4.8mg / kg body weight, 4.9mg / kg body weight, 5.0mg / kg body weight, 5.1mg / kg body weight, 5.2mg / kg body weight, 5.3mg / kg body weight, 5.4mg / kg body weight, 5.5 mg / kg body weight, 5.6 mg / kg body weight, 5.7 mg / kg body weight, 5.8 mg / kg body weight, 5.9 mg / kg body weight, 6.0 mg / kg body weight, 6.1 mg / kg body weight, 6.2 mg / kg body weight, 6.3 mg / kg body weight, 6.4 mg / kg body weight, 6.5 mg / kg body weight Weight, 6.6 mg / kg body weight, 6.7 mg / kg body weight, 6.8 mg / kg body weight, 6.9 mg / kg body weight, 7.0 mg / kg body weight, 7.1 mg / kg body weight, 7.2 mg / kg body weight, 7.3 mg / kg body weight, 7.4 mg / kg body weight, 7.5 mg / kg body weight, 7.6 mg / kg body weight, 7.7mg / kg body weight, 7.8mg / kg body weight, 7.9mg / kg body weight, 8.0mg / kg body weight, 8.1mg / kg body weight, 8.2mg / kg body weight, 8.3mg / kg body weight, 8.4mg / kg body weight, 8.5mg / kg body weight, 8.6mg / kg body weight, 8.7mg / kg body weight, 8.8mg / kg body weight, 8.9mg / kg body weight, 9.0mg / kg body weight, 9.1mg / kg body weight, 9.2mg / kg body weight, 9.3mg / kg body weight, 9.4mg / kg body weight, 9.5mg / kg body weight, 9.6mg / kg body weight, 9.7mg / kg body weight, 9.8mg / kg body weight, 9.9mg / kg body weight Weight, 10.0mg / kg body weight, 10.1mg / kg body weight, 10.2mg / kg body weight, 10.3mg / kg body weight, 10.4mg / kg body weight, 10.5mg / kg body weight, 10.6mg / kg body weight, 10.7mg / kg body weight, 10.8mg / kg body weight, 10.9mg / kg body weight, 11.0mg / kg body weight, 11.1mg / kg body weight, 11.2mg / kg body weight, 11.3mg / kg body weight, 11.4mg / kg body weight, 11.5mg / kg body weight, 11.6mg / kg body weight, 11.7mg / kg body weight, 11.8mg / kg body weight, 11.9mg / kg body weight, 12.0mg / kg body weight g bw, 12.1mg / kg bw, 12.2mg / kg bw, 12.3mg / kg bw, 12.4mg / kg bw, 12.5mg / kg bw, 12.6mg / kg bw, 12.7mg / kg bw, 12.8mg / kg bw, 12.9mg / kg bw, 13.0mg / kg bw, 13.1mg / kg bw, 13.2mg / kg bw, 13.3mg / kg bw, 13.4mg / kg bw, 13.5mg / kg bw, 13.6mg / kg bw, 13.7mg / kg bw, 13.8mg / kg bw, 13.9mg / kg bw, 14.0mg / kg bw, 14.1m g / kg body weight, 14.2mg / kg body weight, 14.3mg / kg body weight, 14.4mg / kg body weight, 14.5mg / kg body weight, 14.6mg / kg body weight, 14.7mg / kg body weight, 14.8mg / kg body weight, 14.9mg / kg body weight, 15.0mg / kg body weight, 15.1mg / kg body weight, 15.2mg / kg body weight, 15.3mg / kg body weight, 15.4mg / kg body weight, 15.5mg / kg body weight, 15.6mg / kg body weight, 15.7mg / kg body weight, 15.8mg / kg body weight, 15.9mg / kg body weight, 16.0mg / kg body weight, 16.1mg / kg body weight, 16.2 mg / kg body weight, 16.3 mg / kg body weight, 16.4 mg / kg or 16.5 mg / kg body weight or about 0.05-0.5 mg / kg body weight, 0.5-1.0 mg / kg body weight, 1.0-1.5 mg / kg body weight, 1.5-2.0 mg / kg body weight, 2.0-2.5 mg / kg body weight, 2.5-3.0 mg / kg body weight, 3.0-3.5 mg / kg body weight, 3.5-4.0 mg / kg body weight, 4.0-4.5 mg / kg body weight, 4.5-5.0 mg / kg body weight, 5.0-5.5 mg / kg body weight, 5.5-6.0 mg / kg body weight, 6.0-6.5 mg / kg body weight, 6.5-7.0 mg / kg body weight, 7.0-7.5 mg / kg body weight, 7.5-8.0 mg / kg body weight , 8.0-8.5 mg / kg body weight, 8.5-9.0 mg / kg body weight, 9.0-9.5 mg / kg body weight, 9.5-10.0 mg / kg body weight, 10.0-10.5 mg / kg body weight, 10.5-11.0 mg / kg body weight, 11.0-11.5 mg / kg body weight, 11.5-12.0 mg / kg body weight, 12.0-12.5 mg / kg body weight, 12.5-13.0 mg / kg body weight, 13.0-13.5 mg / kg body weight, 13.5-14.0 mg / kg body weight, 14.0-14.5 mg / kg body weight, 14.5-15.0 mg / kg body weight, 15.0-15.5 mg / kg body weight, 15.5-16.0 mg / kg or 16.0-16.5 mg / kg body weight.

[0163] 36. The method of any one of embodiments 1 to 34, wherein the conjugate is administered at a dose that is a maximum tolerated dose.

[0164] 37. The method of any one of embodiments 1 to 37, wherein the conjugate is administered every day, every other day, every three days, every four days, every five days, 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.

[0165] 38. The method of any one of embodiments 1 to 38, wherein the conjugate is administered orally, intravenously, subcutaneously, intramuscularly or topically.

[0166] 39. The method of any one of embodiments 1 to 39, wherein said method does not cause unacceptable hyperglycemia in said subject.

[0167] 40. A method for treating an autoimmune disease in a subject, the method comprising administering to the subject an effective amount of a conjugate, wherein the conjugate comprises (i) 765IGF (SEQ ID NO: 2) and (ii) methotrexate.

[0168] 41. A method for monitoring the effectiveness of a treatment for an autoimmune disease in a subject in need thereof using modulation of the frequency of cells expressing IGF-1R, the method comprising:

[0169] a) administering to the subject an effective amount of a conjugate, wherein the conjugate comprises (i) an IGF-1R ligand or a portion or variant thereof, and (ii) a disease modifying agent; and

[0170] b) subsequently determining said frequency of cells expressing IGF-1R in a sample obtained from said subject.

[0171] The following examples are offered by way of illustration and not by way of limitation.

[0172] Examples

[0173] Example 1: Effects of LX-101 on the viability and phenotype of PBMCs from patients with autoimmune diseases

[0174] LX-101 is a conjugate of IGF-1R ligand (SEQ ID NO: 2) and methotrexate, a well-known inhibitor of dihydrofolate reductase, which ultimately inhibits nucleotide synthesis. Without wishing to be bound by theory, LX-101 can deliver methotrexate to cells that overexpress IGF-1R, which are involved in the pathology of autoimmune diseases, and reduce their viability and proliferation.

[0175] Peripheral blood mononuclear cells (PBMCs) frozen from patients diagnosed with lupus, rheumatoid arthritis and scleroderma were obtained from Discovery Life Sciences. A sample was obtained from a 74-year-old white woman diagnosed with systemic lupus erythematosus (SLE) undergoing active treatment with methotrexate, Celebrex, prednisone and Benlysta. Before receiving treatment, a second sample was obtained from a 46-year-old white woman diagnosed with scleroderma (SSc). A third sample was obtained from a 71-year-old white woman diagnosed with active treatment with nabumetone (Nabumetone) for rheumatoid arthritis (RA). All samples were collected after obtaining informed consent.

[0176] PBMCs were thawed, washed, and cultured at 1 × 10 6PBMCs were resuspended in AIM V medium with 5% human AB serum (hAB) at a concentration of 1×10 cells / mL. 6 Cells / well) and treated with 17 μM LX-101 or hydrochloric acid (HCl) as a vehicle control. LX-101 was provided in 10 mM HCl at a concentration of 4 mM (wherein the concentration is expressed as the amount of methotrexate in the drug). The vehicle control group received HCl at the same concentration as the HCl in the LX-101 solution received by the treatment group. PBMCs were incubated at 37°C with 5% CO2 for three days.

[0177] After three days of incubation with LX-101 or vehicle control, PBMCs were blocked with human TruStain FcX Fc receptor blocking solution and stained with a cocktail of antibodies with markers shown in Table 2. Cells were fixed and flow cytometry data of 50,000 live cell events were collected using a BD LSRFortessa X-20 flow cytometer. Data were analyzed using Kaluza software.

[0178] Table 2: Flow cytometry antibodies

[0179] Markers Fluorescent dyes clone Catalog Number supplier CD3 BUV395 SK7 564001 Becton Dickinson CD4 BV421 OKT4 317434 Biolegend CD8 FITC SK1 344704 Baijin Biotechnology Co., Ltd. CD19 BV711 HIB19 302245 Baijin Biotechnology Co., Ltd. CD14 APC HCD14 325607 Baijin Biotechnology Co., Ltd. CD56 BV711 HCD56 318335 Baijin Biotechnology Co., Ltd. CD11C PE-Cy7 3.9 301607 Baijin Biotechnology Co., Ltd. CD123 BV605 6H6 306025 Baijin Biotechnology Co., Ltd. IGF-1R / CD221 PE 1H7 555999 Becton Dickinson Live / Dead APC-eFluor780 not applicable 65-0865-14 eBiosciences

[0180] Analysis of the effects of LX-101 on endogenous PBMC and IGF-1R + Subpopulation effects. Determine relative to live cells, B cells, CD4 + T cells, CD8 + Percentages of T cells, NK cells, and NKT cells. For samples from SLE, RA, and SSc patients, little difference was observed between vehicle control treated cells and LX-101 treated cells for the bulk population of PBMCs ( Figures 1A-1E ).

[0181] However, for IGF-1R + In the presence of leukemia cells, the subpopulations observed in LX-101 treated cells were significantly reduced compared to those observed in vehicle control treated cells.

[0182] IGF-1R was also compared + CD19 + B cell populations, such as Figure 2A For LX-101 treated patient samples, a decrease in IGF-1R + CD19 +The percentage of B cells decreased. In the vehicle control, the B cell population of SLE samples was lower. However, SLE samples were obtained from patients treated with Benlysta (e.g., Belimumab), a B lymphocyte stimulator (BLyS) specific inhibitor, which has been found to increase the total CD19 in the peripheral blood of SLE patients. + B cells decreased. However, CD3 + The number of T cells was not affected. See Jacob et al. (2010) Arthritis & Rheumatism 62:201-210.

[0183] For IGF-1R + group, CD4 + T cell populations and CD8 + T cell populations were compared for SLE, RA, and SSc samples treated with LX-101, especially for those with high IGF-1R in the vehicle control. + CD4 + In RA samples, IGF-1R was observed to be expressed on T cells + CD3 + CD4 + A significant decrease in T cells ( Figure 2B For SLE, RA, and SSc samples treated with LX-101, IGF-1R + CD3 + CD8 + T cell reduction ( Figure 2C ).

[0184] like Figure 2D As seen in Figure 2, in samples treated with LX-101, IGF-1R + CD56 + CD3 - Natural killer (NK) cells were reduced, particularly for samples from SLE and RA patients. Dysregulation of NK cells and the production of NK-derived cytokines is thought to contribute to the pathology of many autoimmune diseases.

[0185] IGF-1R + CD56 + CD3 + Natural killer T (NKT) cells were compared, and a modest reduction in NKT cells was observed in SLE, RA, and SSc samples treated with LX-101, as shown in Figure 2 Figure 2E as shown in .

[0186] Figures 3A-3J IGF-1R from SLE patients is shown+ Figure 2 Flow cytometry data of PBMCs. After LX-101 treatment, CD4 + T cells ( Figure 3A - Vehicle control; Figure 3B -LX-101), CD8 + T cells ( Figure 3C - Vehicle control; Figure 3D -LX-101), B cells ( Figure 3E - Vehicle control; Figure 3F -LX-101), NKT cells ( Figure 3G - Vehicle control; Figure 3H -LX-101) and NK cells ( Fig. 3I - Vehicle control; Figure 3J -LX-101) + The corresponding data from RA patients are shown in Figures 4A-4J Data from SSc patients are shown in Figures 5A-5J middle.

[0187] Data suggest that LX-101 reduces IGF-1R, which is implicated in the pathogenesis of autoimmune diseases including SLE, RA, and SSc + Immune cell populations. In IGF-1R - Few off-target effects were observed in cells, and it was found that in LX-101-treated cells, relative to IGF-1R + B cells, CD4 + T cells, CD8 + The percentages of T cells, NK cells, and NKT cells were decreased.

[0188] Although efforts have been made to ensure accuracy with respect to numbers used (eg, amounts, temperature, etc.), some experimental errors and deviations should be accounted for.

[0189] One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which can be used to practice the subject matter described herein.The present disclosure is in no way limited to the methods and materials described.

[0190] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the subject matter belongs.

[0191] Throughout this specification and claims, unless the context requires otherwise, the words "comprise," "comprises," and "comprising" are used in a non-exclusive sense. It should be understood that the embodiments described herein include "consisting of" and / or "consisting essentially of" embodiments.

[0192] Where a range of values ​​is provided, it is understood that each intervening value between the upper and lower limits of the range (to one tenth of the unit of the lower limit, unless the context clearly indicates otherwise) and any other stated or intervening value within the stated range is encompassed within the invention. Also encompassed are the upper and lower limits of these smaller ranges that may be independently included within smaller ranges, subject to any specifically excluded limits in the stated ranges. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included.

[0193] Many modifications and other embodiments of the invention set forth herein will occur to those skilled in the art having the benefit of the teachings presented in the foregoing description and the associated drawings. Therefore, it should be understood that the invention is not limited to the specific embodiments disclosed, and modifications and other embodiments are intended to be included within the scope of the appended claims. Although specific terms are employed herein, the terms are used only in a generic and descriptive sense and not for purposes of limitation.

Claims

1. A method for treating an autoimmune disease 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.

2. The method of claim 1, wherein the autoimmune disease is selected from the group consisting of: adrenergic drug resistance, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, adrenal autoimmune disease, allergic encephalomyelitis, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inflammatory eye disease, autoimmune neonatal thrombocytopenia, autoimmune neutropenia, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, autoimmune thyroiditis, Behcet's disease, bullous pemphigoid, cardiomyopathy, cardiotomy syndrome, celiac disease-dermatitis, chronic active hepatitis, chronic fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, Churg-Strauss syndrome (Churg-Strauss syndrome), syndrome), cicatricial pemphigoid, CREST syndrome, cold agglutinin disease, Crohn's disease, Cushing's syndrome, cutaneous graft-versus-host disease (GVHD), dense deposit disease, dermatomyositis, essential mixed cryoglobulinemia, fibromyalgia-fibromyositis, glomerulonephritis (e.g., IgA nephropathy), gluten-sensitive enteropathy, Goodpasture's syndrome, Graves' disease, Guillain-Barre disease, hyperthyroidism (e.g., Hashimoto's thyroiditis), idiopathic pulmonary fibrosis, idiopathic Addison's disease, idiopathic thrombocytopenic purpura (ITP), IgA neuropathy, inflammatory arthritis, irritable bowel disease, juvenile arthritis, lichen planus, lichen sclerosus, lupus [e.g.,Systemic lupus erythematosus (SLE), cutaneous lupus, discoid lupus), Ménière's disease, mixed connective tissue disease, morphea, multiple sclerosis, myasthenia gravis, myocarditis, type 1 or immune-mediated diabetes mellitus, neuritis, other endocrine failure, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychondritis, polyendocrinopathy, polyglandular syndrome, polymyalgia rheumatica, polymyositis, post-MI complications, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud's phenomenon, relapsing polychondritis, Reiter's syndrome, rheumatic heart disease, rheumatoid arthritis, sarcoidosis, Sjogren's syndrome, stiff-man syndrome, systemic sclerosis (SSc), Takayasu's arteritis arteritis), temporal arteritis / giant cell arteritis, ulcerative colitis, urticaria, uveitis, uveitic ophthalmia, vasculitis such as dermatitis herpetiformis, vitiligo, and Wegener's granulomatosis.

3. The method of claim 2, wherein the autoimmune disease is selected from the group consisting of rheumatoid arthritis, Graves' disease, SSc, Cushing's syndrome, idiopathic pulmonary fibrosis, SLE, or Crohn's disease.

4. The method of claim 3, wherein the SSc is diffuse cutaneous SSc.

5. The method of any one of claims 1 to 4, wherein the method results in a decrease in the number of cells expressing IGF-1R in the subject.

6. The method of claim 5, wherein the cells expressing IGF-1R are selected from the group consisting of B lymphocytes, T lymphocytes, NK cells and NKT cells.

7. The method according to claim 6, wherein the T lymphocytes are CD4 + or CD8 + .

8. The method of claim 5 or 6, wherein said reduction in the number of cells expressing IGF-1R is measured by flow cytometry or immunohistochemistry.

9. The method of any one of claims 1 to 8, 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).

10. The method of claim 9, wherein the wild-type IGF-1 comprises SEQ ID NO:3, wherein the wild-type insulin comprises SEQ ID NO:10 or 11, and wherein the wild-type IGF-2 comprises SEQ ID NO:

12.

11. The method of any one of claims 1 to 8, 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.

12. The method of claim 11, 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 SEQ ID NO:11, and the variant of wild-type IGF-2 is at least 90% identical to SEQ ID NO:

12.

13. The method of claim 11 or 12, wherein (i) the binding affinity of the variant of wild-type IGF-1 to insulin-like growth factor binding protein (IGFBP) is reduced compared to the binding affinity of wild-type IGF-1 to the IGFBP, or the binding affinity of the variant of wild-type IGF-2 to IGFBP is reduced compared to the binding affinity of wild-type IGF-2 to the IGFBP, and / or (ii) the affinity of the variant of wild-type IGF-1 to the IGF-1R is increased compared to the affinity of wild-type IGF-1 to the IGF-1R, or the affinity of the variant of wild-type IGF-2 to the IGF-1R is increased compared to the affinity of wild-type IGF-2 to the IGF-1R.

14. The method according to any one of claims 1 to 13, wherein the IGF-1R ligand, or a portion or variant thereof, comprises a leader sequence. The method of claim 14 , wherein the leader sequence comprises SEQ ID NO:

1.

16. The method of any one of claims 1 to 8 and claims 11 to 15, 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).

17. The method of claim 16, wherein the IGF-1R ligand comprises 765IGF (SEQ ID NO: 2).

18. The method of any one of claims 1 to 17, wherein the disease-modifying agent comprises a cytotoxic agent.

19. The method of claim 18, wherein the cytotoxic agent comprises a chemotherapeutic agent.

20. The method according to claim 19,wherein the chemotherapeutic agent is amsacrine, azacytidine, 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 C), mitotane, mitoxantrone, oxaliplatin, paclitaxel, pemetrexed, pentostatin, plicamycin, procarbazine, raltitrexed, semustine, streptozocin, temozolamide, teniposide, thioguanine, thiotepa, topotecan, trimetrexate, valrubicin, vincristine, vinblastine, vindesine, or vinorelbine.

21. The method of claim 20, wherein the chemotherapeutic agent is methotrexate.

22. The method of claim 18, wherein the cytotoxic agent comprises a toxin.

23. The method of claim 22, wherein the toxin comprises Clostridium perfringens enterotoxin, diphtheria toxin, ricin chain A, Pseudomonas exotoxin, A chain toxin, ribosome inactivating protein, alpha-sarcin, aspergillus, or ribonuclease.

24. The method of claim 23, wherein the toxin comprises Clostridium perfringens enterotoxin or a portion or variant thereof.

25. The method of claim 24, wherein the conjugate comprises SEQ ID NO: 14 or SEQ ID NO:

15.

26. The method of claim 23, wherein the toxin comprises diphtheria toxin or a portion or variant thereof.

27. The method of claim 26, wherein the toxin comprises SEQ ID NO: 13 or SEQ ID NO:

16.

28. The method of any one of claims 1 to 27, wherein the subject (i) has not previously received treatment for the autoimmune disease; (ii) has previously received treatment for the autoimmune disease; (iii) has relapsed from a previous treatment for the autoimmune disease; or (iv) is refractory to a previous treatment for the autoimmune disease.

29. The method of any one of claims 1 to 28, wherein the conjugate is administered in combination with one or more other therapies.

30. The method of claim 29, wherein the one or more additional therapies comprise prednisone, hydroxychloroquine, chloroquine, belimumab, anifrolumab, abatacept, atacicept, lupuzor, rituximab, voclosporin, aldesleukin, baricitinib, BIIB059, BI655064, bortezomib, BT063, cenerimod, dapirolizumab pegol.pegol), edratide, filgotinib, GS-9876, iberdomide, IFN-α kinin, iguratimod, nelfinavir, obinutuzumab, OMS721, rapamycin, RC18, RSLV-132, SM101, theralizumab, ustekinumab ), vobarilizumab, XmAb5871, blisibimod, tabalumab, epratuzumab, rigerimod, tacrolimus, rontalizumab, sifalimumab, anifrolumab, tocilizumab, infliximab iximab), metelimumab, fresolimumab, rilonacept, cyclophosphamide, methotrexate, nintedanib, JBT-101, imatinib, pirfenidone, nilotinib, dasatinib, SAR100842, BMS-986202, BAY41-2272, riociguat guat, resunab, ixekizumab, brodalumab, tralokinumab, etanercept, adalimumab, golimumab, secukinumab, tildrakizumab, tofacitinib, or guselkumab.

31. The method of any one of claims 1 to 30, wherein the conjugate is administered at about 0.05 μEq / kg body weight, 0.10 μEq / kg body weight, 0.20 μEq / kg body weight, 0.40 μEq / kg body weight, 0.80 μEq / kg body weight, 1.0 μEq / kg body weight, 1.5 μEq / kg body weight, 1.6 μEq / kg body weight, 2.0 μEq / kg body weight, 2.5 μEq / kg body weight, 3.0 μEq / kg body weight, , 3.5 μEq / kg body weight, 4.0 μEq / kg body weight, 4.5 μEq / kg body weight, 5.0 μEq / kg body weight, 5.5 μEq / kg body weight, 6.0 μEq / kg body weight, 6.5 μEq / kg body weight, 7.0 μEq / kg body weight, 7.5 μEq / kg body weight, 8.0 μEq / kg body weight, 8.5 μEq / kg body weight, 9.0 μEq / kg body weight, 9.5 μEq / kg body weight, or 10.0 μEq / kg The dosage of body weight is about 0.05-0.5 μEq / kg body weight, 0.5-1.0 μEq / kg body weight, 1.0-1.5 μEq / kg body weight, 1.5-2.0 μEq / kg body weight, 2.0-2.5 μEq / kg body weight, 2.5-3.0 μEq / kg body weight, 3.0-3.5 μEq / kg body weight, 3.5-4.0 μEq / kg body weight, 4.0-4.5 μEq / kg body weight, 4.5-5.0 μEq / kg body weight, A dose range of 5.0-5.5 μEq / kg body weight, 5.5-6.0 μEq / kg body weight, 6.0-6.5 μEq / kg body weight, 6.5-7.0 μEq / kg body weight, 7.0-7.5 μEq / kg body weight, 7.5-8.0 μEq / kg body weight, 8.0-8.5 μEq / kg body weight, 8.5-9.0 μEq / kg body weight, 9.0-9.5 μEq / kg body weight or 9.5-10.0 μEq / kg body weight is administered.

32. according to the method of any one of claims 1 to 31, wherein the conjugate is at about 0.05mg / kg body weight, 0.10mg / kg body weight, 0.15mg / kg body weight, 0.20mg / kg body weight, 0.25mg / kg body weight, 0.30mg / kg body weight, 0.35mg / kg body weight, 0.40mg / kg body weight, 0.45mg / kg body weight, 0.50mg / kg body weight, 0.55mg / kg body weight, 0.60mg / kg body weight, 0.65mg / kg body weight, 0.70mg / kg body weight, 0.75mg / kg body weight, 0.80mg / kg body weight, 0.85mg / kg body weight, 0.90mg / kg body weight, 0. .95mg / kg body weight, 1.0mg / kg body weight, 1.1mg / kg body weight, 1.2mg / kg body weight, 1.3mg / kg body weight, 1.4mg / kg body weight, 1.5mg / kg body weight, 1.6mg / kg body weight, 1.7mg / kg body weight, 1.8mg / kg body weight, 1.9mg / kg body weight, 2.0m g / kg body weight, 2.1 mg / kg body weight, 2.2 mg / kg body weight, 2.3 mg / kg body weight, 2.4 mg / kg body weight, 2.5 mg / kg body weight, 2.6 mg / kg body weight, 2.7 mg / kg body weight, 2.8 mg / kg body weight, 2.9 mg / kg body weight, 3.0 mg / kg body weight, 3.1 mg / kg Body weight, 3.2 mg / kg body weight, 3.3 mg / kg body weight, 3.4 mg / kg body weight, 3.5 mg / kg body weight, 3.6 mg / kg body weight, 3.7 mg / kg body weight, 3.8 mg / kg body weight, 3.9 mg / kg body weight, 4.0 mg / kg body weight, 4.1 mg / kg body weight, 4.2 mg / kg body weight, 4 .3mg / kg body weight, 4.4mg / kg body weight, 4.5mg / kg body weight, 4.6mg / kg body weight, 4.7mg / kg body weight, 4.8mg / kg body weight, 4.9mg / kg body weight, 5.0mg / kg body weight, 5.1mg / kg body weight, 5.2mg / kg body weight, 5.3mg / kg body weight, 5.4mg / kg body weight, 5.5 mg / kg body weight, 5.6 mg / kg body weight, 5.7 mg / kg body weight, 5.8 mg / kg body weight, 5.9 mg / kg body weight, 6.0 mg / kg body weight, 6.1 mg / kg body weight, 6.2 mg / kg body weight, 6.3 mg / kg body weight, 6.4 mg / kg body weight, 6.5 mg / kg body weight Weight, 6.6 mg / kg body weight, 6.7 mg / kg body weight, 6.8 mg / kg body weight, 6.9 mg / kg body weight, 7.0 mg / kg body weight, 7.1 mg / kg body weight, 7.2 mg / kg body weight, 7.3 mg / kg body weight, 7.4 mg / kg body weight, 7.5 mg / kg body weight, 7.6 mg / kg body weight, 7.7mg / kg body weight, 7.8mg / kg body weight, 7.9mg / kg body weight, 8.0mg / kg body weight, 8.1mg / kg body weight, 8.2mg / kg body weight, 8.3mg / kg body weight, 8.4mg / kg body weight, 8.5mg / kg body weight, 8.6mg / kg body weight, 8.7mg / kg body weight, 8.8mg / kg body weight, 8.9mg / kg body weight, 9.0mg / kg body weight, 9.1mg / kg body weight, 9.2mg / kg body weight, 9.3mg / kg body weight, 9.4mg / kg body weight, 9.5mg / kg body weight, 9.6mg / kg body weight, 9.7mg / kg body weight, 9.8mg / kg body weight, 9.9mg / kg body weight Weight, 10.0mg / kg body weight, 10.1mg / kg body weight, 10.2mg / kg body weight, 10.3mg / kg body weight, 10.4mg / kg body weight, 10.5mg / kg body weight, 10.6mg / kg body weight, 10.7mg / kg body weight, 10.8mg / kg body weight, 10.9mg / kg body weight, 11.0mg / kg body weight, 11.1mg / kg body weight, 11.2mg / kg body weight, 11.3mg / kg body weight, 11.4mg / kg body weight, 11.5mg / kg body weight, 11.6mg / kg body weight, 11.7mg / kg body weight, 11.8mg / kg body weight, 11.9mg / kg body weight, 12.0mg / kg body weight g bw, 12.1mg / kg bw, 12.2mg / kg bw, 12.3mg / kg bw, 12.4mg / kg bw, 12.5mg / kg bw, 12.6mg / kg bw, 12.7mg / kg bw, 12.8mg / kg bw, 12.9mg / kg bw, 13.0mg / kg bw, 13.1mg / kg bw, 13.2mg / kg bw, 13.3mg / kg bw, 13.4mg / kg bw, 13.5mg / kg bw, 13.6mg / kg bw, 13.7mg / kg bw, 13.8mg / kg bw, 13.9mg / kg bw, 14.0mg / kg bw, 14.1m g / kg body weight, 14.2mg / kg body weight, 14.3mg / kg body weight, 14.4mg / kg body weight, 14.5mg / kg body weight, 14.6mg / kg body weight, 14.7mg / kg body weight, 14.8mg / kg body weight, 14.9mg / kg body weight, 15.0mg / kg body weight, 15.1mg / kg body weight, 15.2mg / kg body weight, 15.3mg / kg body weight, 15.4mg / kg body weight, 15.5mg / kg body weight, 15.6mg / kg body weight, 15.7mg / kg body weight, 15.8mg / kg body weight, 15.9mg / kg body weight, 16.0mg / kg body weight, 16.1mg / kg body weight, 16.2 mg / kg body weight, 16.3 mg / kg body weight, 16.4 mg / kg body weight or 16.5 mg / kg body weight or about 0.05-0.5 mg / kg body weight, 0.5-1.0 mg / kg body weight, 1.0-1.5 mg / kg body weight, 1.5-2.0 mg / kg body weight, 2.0-2.5 mg / kg body weight, 2.5-3.0 mg / kg body weight, 3.0-3.5 mg / kg body weight, 3.5-4.0 mg / kg body weight, 4.0-4.5 mg / kg body weight, 4.5-5.0 mg / kg body weight, 5.0-5.5 mg / kg body weight, 5.5-6.0 mg / kg body weight, 6.0-6.5 mg / kg body weight, 6.5-7.0 mg / kg body weight, 7.0-7.5 mg / kg body weight, 7.5-8.0 mg / kg body weight , 8.0-8.5 mg / kg body weight, 8.5-9.0 mg / kg body weight, 9.0-9.5 mg / kg body weight, 9.5-10.0 mg / kg body weight, 10.0-10.5 mg / kg body weight, 10.5-11.0 mg / kg body weight, 11.0-11.5 mg / kg body weight, 11.5-12.0 mg / kg body weight, 12.0-12.5 mg / kg body weight, 12.5-13.0 mg / kg body weight, 13.0-13.5 mg / kg body weight, 13.5-14.0 mg / kg body weight, 14.0-14.5 mg / kg body weight, 14.5-15.0 mg / kg body weight, 15.0-15.5 mg / kg body weight, 15.5-16.0 mg / kg body weight or 16.0-16.5 mg / kg body weight.

33. The method of any one of claims 1 to 32, wherein the conjugate is administered at a dose that is the maximum tolerated dose.

34. The method of any one of claims 1 to 33, wherein the conjugate is administered every day, every other day, every three days, every four days, every five days, 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.

35. The method of any one of claims 1 to 34, wherein the conjugate is administered orally, intravenously, subcutaneously, intramuscularly or topically.

36. The method of any one of claims 1 to 8 and 11 to 35, wherein the IGF-1R ligand is SEQ ID NO: 2, the cytotoxic agent is methotrexate, and wherein there are 6 to 10 methotrexate molecules per IGF-1R ligand of SEQ ID NO:

2.

37. The method of claim 36, wherein the autoimmune disease is SLE.

38. The method of claim 36, wherein the autoimmune disease is RA.

39. The method of claim 36, wherein the autoimmune disease is SSc.

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