Superagonists, partial agonists and antagonists of interleukin-2
Inventors
GARCIA, Christopher K. • MITRA, Suman • Leonard, Warren J. • RING, Aaron M.
Assignees
National Institutes of Health NIH • Leland Stanford Junior University
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Publication Number
US-12202873-B2
Publication Date
2025-01-21
Expiration Date
2035-04-24
Abstract
Novel human interleukin-2 (IL-2) muteins or variants thereof are provided. In particular, provided are IL-2 muteins that have an increased binding capacity for IL-2Rβ receptor and a decreased binding capacity for IL-2Rγc receptor, as compared to wild-type IL-2. Such IL-2 muteins are useful, for example, as IL-2 partial agonist and antagonists in applications where reduction or inhibition of one or more IL-2 and/or IL-15 functions is useful (e.g., in the treatment of graft versus host disease (GVHD) and adult T cell leukemia). Also provided are nucleic acids encoding such IL-2 muteins, methods of making such IL-2 muteins, pharmaceutical compositions that include such IL-2 muteins and methods of treatment using such pharmaceutical compositions.
Core Innovation
Novel human interleukin-2 (IL-2) muteins or variants thereof are provided, particularly those having an increased binding capacity for the IL-2Rβ receptor and a decreased binding capacity for the IL-2Rγc receptor compared to wild-type IL-2. These IL-2 muteins are useful as IL-2 partial agonists and antagonists, offering potential therapeutic applications where reduction or inhibition of one or more IL-2 and/or IL-15 functions is desirable, including treatments for graft versus host disease (GVHD) and adult T cell leukemia. Additionally, nucleic acids encoding such IL-2 muteins, pharmaceutical compositions containing them, and methods of treatment using these compositions are provided.
IL-2 plays a crucial role in the immune response by promoting proliferation and expansion of activated T lymphocytes, potentiating B cell growth, and activating monocytes and natural killer (NK) cells. IL-2 signals through three receptors: IL-2Rα, IL-2Rβ, and IL-2Rγc. Cells such as resting T cells express IL-2Rβ and IL-2Rγc, which exhibit low affinity for IL-2. Upon stimulation, these cells express IL-2Rα, which increases receptor affinity and leads to T cell activation. Existing immunomodulatory agents like monoclonal antibodies targeting IL-2Rα do not block all IL-2 signaling pathways, particularly those via the IL-2Rβγc intermediate affinity receptors, nor IL-15 signaling, limiting their therapeutic efficacy.
The invention addresses the need for IL-2 muteins capable of blocking one or more IL-2 and/or IL-15 functions. Prior developed IL-2 “superkines” with augmented IL-2Rβ binding affinity were hypothesized to serve as dominant-negative scaffolds to create a receptor signaling clamp. By directed mutation of these super-IL-2 full agonists to diminish their binding to IL-2Rγc, the resulting partial agonists and non-signaling (neutral) molecules attenuate IL-2Rβ-γc heterodimerization, offering a new class of IL-2 antagonists that block endogenous cytokines without eliciting their own biological activity.
Claims Coverage
The patent contains several independent claims focusing on nucleic acids encoding IL-2 muteins with modified binding affinities, and fusion proteins comprising such muteins linked to other polypeptides.
IL-2 mutein with increased IL-2Rβ affinity and decreased IL-2Rγc affinity
A nucleic acid encoding an IL-2 mutein comprising amino acid substitutions L18R, Q22E, Q126T, and S130R, which exhibits increased binding affinity for IL-2Rβ and decreased binding affinity for IL-2Rγc compared to wild-type IL-2.
Additional amino acid substitutions increasing IL-2Rβ binding affinity
The IL-2 mutein further comprises one or more amino acid substitutions selected from Q74N, Q74H, Q74S, L80F, L80V, R81D, R81T, L85V, I86V, I89V, and I92V that enhance IL-2Rβ binding affinity.
Specific IL-2 mutein substitution combinations enhancing IL-2Rβ affinity
IL-2 muteins comprising combinations such as L80F, R81D, L85V, I86V, and I92F; Q74N, L80F, R81D, L85V, I86V, I89V, and I92F; and others are claimed as nucleic acids encoding these muteins.
Reduced signaling capability in IL-2Rβ+ T cells
The muteins possess decreased capabilities to stimulate STAT5 phosphorylation and pERK1/ERK2 signaling in IL-2Rβ+ cells compared to wild-type IL-2.
Inhibition of IL-2 induced cellular functions
The muteins act as inhibitors of IL-2 STAT5 phosphorylation, proliferation of CD8+ T cells, TCR-induced cell proliferation, Th1, Th9 and Treg differentiation; and as promoters of Th17 differentiation and inhibitors of IL-2 dependent activation of NK cells.
IL-2 mutein fusion proteins
Nucleic acids encoding fusion proteins comprising the IL-2 muteins linked to human Fc antibody fragments or other heterologous polypeptides, including albumin, are claimed.
The claims cover nucleic acids encoding IL-2 muteins with specific amino acid substitutions that enhance IL-2Rβ binding and diminish IL-2Rγc binding, thus functioning as partial agonists or antagonists of IL-2. Further claims include fusion proteins with these muteins linked to Fc domains or other polypeptides and describe muteins' reduced signaling and inhibitory effects on IL-2 and IL-15 mediated functions.
Stated Advantages
IL-2 muteins function as partial agonists and antagonists, allowing selective modulation of IL-2 and IL-15 functions.
Enhanced binding affinity for IL-2Rβ coupled with decreased IL-2Rγc binding creates a receptor signaling clamp that blocks endogenous cytokine signaling.
Potential therapeutic benefits include treatment of graft versus host disease (GVHD) and adult T-cell leukemia with reduced adverse effects compared to wild-type IL-2.
The muteins have reduced vascular leak syndrome and toxicity in vivo, improving safety profiles.
IL-2 muteins promote desired immune responses such as enhanced memory phenotype T cell expansion and anti-tumor activity while limiting expansion of suppressive regulatory T cells.
Documented Applications
Treatment of graft versus host disease (GVHD) by administering IL-2 muteins that inhibit IL-2 and/or IL-15 signaling.
Treatment of adult T-cell leukemia (ATL), particularly chronic and smoldering forms, using IL-2 mutein antagonists that inhibit cytokine-dependent malignant T cell proliferation.
Use as immunosuppressive agents to selectively dampen IL-2 and IL-15 dependent immune activation and T cell proliferation.
Therapeutic application in cancer treatment by enhancing anti-tumor responses through IL-2 muteins with enhanced IL-2Rβ binding and reduced toxicity.
Ex vivo immune cell modulation, such as expansion or inhibition of T cell populations, by culturing cells with IL-2 muteins before administration.
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