Methods of treating an autoimmune disease with a human interleukin-3 (IL-3)-diphtheria toxin conjugate (DT-IL3)
The present disclosure provides, in part, a method of treating an autoimmune disease in a subject by reducing the number of pDCs through administration of a human interleukin-3 (IL-3)-diphtheria toxin conjugate (DT-IL3). The disclosure also generally relates to methods of monitoring the effectiveness of therapy in subjects receiving DT-IL3 for treating an autoimmune disease, and methods of determining continuing treatment of subjects receiving DT-IL3 for treating an autoimmune disease. The disclosure also provides pharmaceutical compositions of DT-IL3 for use in such methods.
1 . A method of treating an autoimmune disease in a human subject, the method comprising reducing the number of plasmacytoid dendritic cells (pDCs) in the subject by administering a therapeutically effective dose of a human interleukin-3 (IL-3)-diphtheria toxin conjugate (DT-IL3) to the human subject,
wherein the DT-IL3 comprises diphtheria toxin (DT) and interleukin-3 protein (IL-3), linked via a linker molecule, wherein:
(a) the DT comprises an amino acid sequence having a sequence at least 98% identical to the amino acid sequence of SEQ ID NO:3, wherein the amino acid sequence at least 98% identical to the amino acid sequence of SEQ ID NO: 3 comprises at least two domains of DT comprising:
(i) a catalytic domain comprising the sequence of amino acids 26-112 of SEQ ID NO: 1; and
(ii) a translocation domain comprising amino acids 225-404 of SEQ ID NO:1, and
(b) the IL-3 comprises an amino acid sequence having a sequence at least 95% identical to a mature IL-3 amino acid sequence having the amino acid sequence of SEQ ID NO: 2 without a signal peptide;
wherein the human subject prior to treatment has an elevated expression of at least one type I IFN-inducible gene in the skin or peripheral blood as compared to a healthy human subject; and
wherein the treatment reduces the level of at least one cytokine or chemokine selected from chemokine ligand 4 (CXCL4), CXCL9, CXCL10, one or more type I interferons (type I IFNs), interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α), IL-8, regulated upon activation normal T cell expressed and presumably secreted (RANTES), macrophage inflammatory protein 1α (MIP1α), MIP1β, and monocyte chemoattractant protein-1 (MCP1) in the human subject.
2 . The method of claim 1 , wherein the reduction in the number of pDCs is accomplished through killing of pDCs by the administered DT-IL3 without affecting the number of T cells or B cells in the human subject by more than 15% as compared to the number of T cells or B cells in the human subject before administration of DT-IL3.
3 . The method of claim 1 , wherein the autoimmune disease is selected from lupus, Sjogren's syndrome, inflammatory arthritis, systemic sclerosis (SSc), morphea, psoriasis, lichen planus, dermatomyositis, lichen sclerosus, cutaneous graft-versus-host disease (GVHD), adrenergic drug resistance, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, autoimmune diseases of the adrenal gland, allergic encephalomyelitis, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inflammatory eye disease, autoimmune neonatal thrombocytopenia, autoimmune neutropenia, autoimmune oophoritis, autoimmune orchitis, autoimmune thrombocytopenia, autoimmune thyroiditis, Behcet's disease, bullous pemphigoid, cardiomyopathy, cardiotomy syndrome, celiac sprue-dermatitis, chronic active hepatitis, chronic fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, Churg-Strauss syndrome, cicatricial pemphigoid, CREST syndrome, cold agglutinin disease, Crohn's disease, dense deposit disease, essential mixed cryoglobulinemia, fibromyalgia-fibromyositis, glomerulonephritis, gluten-sensitive enteropathy, Goodpasture's syndrome, Graves' disease, Guillain-Barre, hyperthyroidism, idiopathic pulmonary fibrosis, idiopathic Addison's disease, idiopathic thrombocytopenia purpura (ITP), IgA neuropathy, juvenile arthritis, Ménière's disease, mixed connective tissue disease, multiple sclerosis, Myasthenia Gravis, myocarditis, type 1 diabetes mellitus, neuritis, endocrine gland failure, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychondritis, polyendocrinopathies, polyglandular syndromes, polymyalgia rheumatica, polymyositis, post-myocardial infarction (post-MI), primary agammaglobulinemia, primary biliary cirrhosis, psoriatic arthritis, Raynaud's phenomenon, relapsing polychondritis, Reiter's syndrome, rheumatic heart disease, rheumatoid arthritis, sarcoidosis, stiff-man syndrome, takayasu arteritis, temporal arteritis/giant cell arteritis, ulcerative colitis, urticaria, uveitis, Uveitis Ophthalmia, vasculitides, vitiligo, and Wegener's granulomatosis.
4 . The method of claim 1 , wherein the type I IFN-inducible gene is selected from IFI27, IFI30, IFI6, IRF1, IFITM1, IFITM2, IFIT1, CXCL9, CXCL10, RSAD2, IFI44, IFI44L, IFI6, GBP1, MxB, SERPING1, LY6E, XAF3, IFIT2, IFIT3, MxA, and IRF7.
5 . The method of claim 3 , wherein the human subject exhibits elevated levels of one or more type I interferons compared to healthy individuals.
6 . The method of claim 1 , wherein the reduction in the number of pDCs results in reduced levels of at least one type I IFN-inducible gene as compared to levels of the at least one type I IFN-inducible gene in the human subject before administration of DT-IL3.
7 . The method of claim 1 , wherein the therapeutically effective dose of DT-IL3 is in the range of 1 μg/kg to 100 μg/kg.
8 . The method of claim 7 , wherein the administering of the therapeutically effective dose of DT-IL3 is repeated by administering the DT-IL3 for treatment cycles of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks.
9 . The method of claim 8 , wherein the administering of the therapeutically effective dose of DT-IL3 is repeated by administering the DT-IL3 at 1, 2, 3, 4, or 5 times per treatment cycle.
10 . The method of claim 9 , wherein the administering of the therapeutically effective dose of DT-IL3 is once daily for 1 day per treatment cycle, or once daily for 2, 3, 4, or 5 consecutive days per treatment cycle.
11 . The method of claim 7 , wherein the administration of the therapeutically effective dose of DT-IL3 does not affect the number of T cells or B cells in the subject.
12 . The method of claim 1 , wherein the number of pDCs in the human subject after administration of DT-IL3 is reduced by about 20% to about 95% as compared to the number of pDCs in the human subject before administration of DT-IL3.
13 . The method of claim 1 , wherein the type I interferon is IFN-α.
14 . The method of claim 1 ,
wherein the DT comprises an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO:3, and
wherein IL-3 comprises an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO:2 without the signal peptide.
15 . The method of claim 9 , wherein the treatment cycle of 1, 2, 3, 4, 5, 6, 7, or 8 weeks is repeated 1, 2, 3, 4, 5 or more times.
16 . The method of claim 14 ,
wherein the DT comprises the amino acid sequence of SEQ ID NO:3, and
wherein IL-3 comprises the amino acid sequence of SEQ ID NO:2 without the signal peptide.