Interleukin-2 variants and methods of uses thereof
Inventors
Li, Yue-Sheng • Rui, Lingyun • Xu, Jing
Assignees
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Publication Number
US-12152060-B2
Publication Date
2024-11-26
Expiration Date
Abstract
The present invention relates to polypeptides which share primary sequence with human Interleukin 2 (IL-2), except for several amino acids that have been mutated. One panel of IL-2 variants comprise mutations with impressive manufacturability that preferentially promotes the proliferation, survival, activation and/or function of immunosuppressive regulatory T cells (Tregs) (Treg: CD4+CD25+FoxP3+) over effector T cells and Natural Killer Cells (NK). cells. Also includes therapeutic uses of such IL-2 selective agent, used alone, or in combination with immune modulating agents or disease-tissue targeting antibody, protein or peptide to treat Treg cell-deficiency, various autoimmune and inflammatory disorders, organ transplantation and graft-versus-host disease. In another aspect the present invention relates to pharmaceutical compositions comprising the polypeptides disclosed. Finally, the present invention relates to the therapeutic use of the polypeptides and pharmaceutical compositions disclosed due to their selective modulating effect of the immune system on diseases like autoimmune and inflammatory disorders.
Core Innovation
Engineered interleukin-2 (IL-2) variants and IL-2 Fc fusion proteins are disclosed to address IL-2 developability and selectivity issues. The disclosure reports IL-2 variants that preferentially signal through the high-affinity IL-2Rαβγ pathway while reducing or ablating interaction with the intermediate-affinity IL-2Rβγc pathway, thereby favoring expansion and function of CD4+CD25+FOXP3+ regulatory T cells (Tregs) over effector T cells and NK cells.
The disclosure further describes design rationale including targeting IL-2Rβ/γc interface residues and removing a proposed “LDL” toxin-like motif, including D20 and flanking residues, to reduce vascular toxicity risk (VLS). It also describes an IL-2 position 125 substitution, S125I (Ile-125), as a broadly beneficial engineering change with higher expression and uniformly low SEC aggregation across multiple mutational contexts, while generally retaining IL-2 biological activity and Treg selectivity.
Additional constructs are described in fusion formats that include Fc or other heterologous half-life-extending carriers. The disclosure emphasizes carrier fusion configurations including linker/hinge and Fc engineering to reduce effector functions and extend half-life, and it includes therapeutic compositions and nucleic acids/vectors encoding the IL-2 variants as part of the overall inventive concept.
Claims Coverage
The provided independent claims cover two inventive features: an IL-2 variant fusion protein with defined sequence components and an Fc domain linked by a defined peptide linker, and a pharmaceutical composition containing that fusion protein. The claim set also includes a two-polypeptide chain architecture for one fusion protein, connected by interchain disulfide bonds.
Il-2 variant fused to Fc via a defined peptide linker
An isolated fusion protein comprising an interleukin-2 (IL-2) variant polypeptide having the amino acid sequence set forth in SEQ ID NO: 193, fused to an Fc domain having the amino acid sequence set forth in SEQ ID NO: 45, wherein the IL-2 variant polypeptide is fused to the Fc domain by a peptide linker having the amino acid sequence set forth in SEQ ID NO: 55.
Pharmaceutical composition with an isolated IL-2 Fc fusion protein and a pharmaceutically acceptable carrier
A pharmaceutical composition comprising an isolated fusion protein having the amino acid sequence set forth in SEQ ID NO: 198 in admixture with a pharmaceutically acceptable carrier.
Overall, the independent claims define an IL-2 Fc fusion protein with specified IL-2 variant and Fc sequences connected by a specified peptide linker, and extend coverage to a pharmaceutical composition formulated with the isolated fusion protein and a pharmaceutically acceptable carrier. The dependent claims further emphasize a two-polypeptide chain structure with interchain disulfide bonds.
Stated Advantages
Preferential signaling through the high-affinity IL-2Rαβγ pathway while reducing or ablating intermediate-affinity IL-2Rβγc interaction.
Bias expansion and function of CD4+CD25+FOXP3+ regulatory T cells (Tregs) over effector T cells and NK cells.
Reduce vascular toxicity risk (VLS) by removing a proposed “LDL” toxin-like motif, including D20 and flanking residues.
Higher expression and uniformly low SEC aggregation across multiple mutational contexts for S125I (Ile-125) constructs.
Reduced or ablated Fc effector functions and extended half-life for Fc fusion formats.
Generally retained IL-2 biological activity and Treg selectivity for the described S125I (Ile-125) constructs.
Reduced IL-2Rβ/γc interaction as indicated by receptor-complex binding data.
Preferential expansion or proliferation of Treg cells in vivo.
Reduced activity on effector and NK lineages, including dose-response and repeated-dosing effects.
Suppression of antigen-driven inflammation in a DTH mouse model.
Documented Applications
Therapeutic use for autoimmune/inflammatory disorders.
Therapeutic use for organ transplantation/GvHD.
Therapeutic use includes pharmaceutical compositions comprising the IL-2 variant fusion proteins.
Therapeutic use includes nucleic acids/vectors encoding the IL-2 variants.
Preferential expansion/proliferation of Treg cells with reduced activity on effector and NK lineages in mouse studies, including effects described with dose-response and repeated-dosing.
Suppression of antigen-driven inflammation in a DTH mouse model (KLH-induced).
Functional evaluation of IL-2 biological activity and selectivity using human PBMC assays and associated receptor-complex binding data.
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