IL-2 Mutant Design for Selective Regulatory T-Cell Activation
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Solution Overview
Problem
Existing IL-2 drugs face challenges in stability and yield, and their binding abilities to specific receptor complexes are not optimized for preferential stimulation of regulatory T cells over non-regulatory T cells and NK cells.
Innovation Solution
Development of IL-2 mutants with specific mutations at positions Q13, L18, G27, Y31, A73, H79, P82, I89, N90, V91, V93, and R120, which reduce binding to IL-2Rβγ subunits and enhance preferential stimulation of regulatory T cells through STAT5 phosphorylation and proliferation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If wild-type IL-2 is used to activate immune cells, then both non-regulatory T cells and regulatory T cells are activated, but non-specific stimulation occurs and selectivity is reduced
Solution Approach 1:
The patent applies local quality by introducing specific point mutations at defined positions (Q13, L18, G27, Y31, A73, H79, P82, I89, N90, V91, V93, R120) in the IL-2 molecule. These localized changes modify the binding interface to preferentially recognize IL-2Rαβγ complexes on regulatory T cells while reducing affinity for IL-2Rβγ on non-regulatory cells, thereby achieving selectivity without altering the overall IL-2 structure
Solution Approach 2:
The patent employs parameter changes by systematically varying amino acid residues at 12 specific positions to optimize binding characteristics. Mutations such as Q13L, L18I, G27W, Y31V, A73L, H79Q, P82L, I89L, N90Y, V91A, V93I, F117W, and R120F alter the physical-chemical properties of the IL-2 molecule, enabling preferential binding to high-affinity IL-2Rαβγ complexes while reducing interaction with low-affinity IL-2Rβγ complexes
2Reliability
If high doses of IL-2 are administered to treat tumors, then immune cell activation is enhanced, but stability and yield of the drug are insufficient
Solution Approach 1:
The patent applies parameter changes by introducing mutations specifically targeted at improving stability (Q13L, L18I, G27W, Y31V, A73L, H79Q, P82L, I89L, N90Y, V91A, V93I, F117W, R120F) and yield (H16E, D20A, D20H, D20Y, N88A, N88I, N88G, N88R, N88D). These amino acid substitutions modify the protein's physical properties, including thermal stability, solubility, and expression efficiency, thereby simultaneously improving both stability and productivity parameters
3Adaptability or versatility
If IL-2 binding ability to IL-2Rβγ subunits is high, then non-regulatory T cells and NK cells are activated, but preferential stimulation of regulatory T cells is reduced
Solution Approach 1:
The patent applies local quality by modifying specific residues (Q13, L18, G27, Y31, A73, H79, P82, I89, N90, V91, V93, R120) that are involved in binding to IL-2Rβγ subunits. These localized changes reduce affinity for βγ complexes while preserving or enhancing affinity for αβγ complexes, thereby achieving preferential stimulation of regulatory T cells through differential binding characteristics
Data Source
AI summary
The present disclosure discloses IL-2 mutants and uses thereof. More specifically, the disclosure provides IL-2 mutants and corresponding fusion proteins, conjugates, nucleic acid fragments, vectors, host cells, methods for preparing the mutants or fusion proteins, IL-2 mutants or fusion proteins prepared according to the methods, pharmaceutical compositions, pharmaceutical uses, methods for treating diseases, and methods for preferentially stimulating regulatory T cells. Compared to wild-type IL-2, the IL-2 mutants of the present disclosure have higher Tm values and improved stability; alternatively, the IL-2 mutants of the present disclosure have an increased yield or changed binding activity to the IL-2Rβγ complexes compared to wild-type IL-2.