Interferon-gamma biased agonists

Inventors

Garcia, Kenan Christopher • Mendoza, Juan Luis

Assignees

Leland Stanford Junior University

Abstract

Disclosed herein are compositions and methods for modulating IFN-γ-mediated signaling by completely or partially agonizing the downstream signal transduction mediated through at least one of the IFN-γ receptors. More particularly, the disclosure provides novel IFN-γ polypeptide variants with reduced binding affinity to at least one of its receptors. The disclosure also provides compositions and methods useful for producing such molecules, as well as methods for the treatment of health diseases associated with the perturbation of signal transduction mediated by IFN-γ.

Core Innovation

The disclosed invention provides IFN-b3 biased/partial agonist polypeptides engineered to reduce binding affinity to one or both IFN-b3 receptors, IFN-b3R1 and/or IFN-b3R2, while retaining downstream signaling. The work focuses on creating IFN-b3 agonist variants with altered receptor binding, characterized relative to an IFN-b3 polypeptide sequence (SEQ ID NO: 1).

Structural determination of the hexameric 2:2:2 IFN-b3/IFN-b3R1/IFN-b3R2 complex is used to identify IFN-b3 residues at the IFN-b3R2 binding interface. Mutations at these positions reduce receptor-binding affinity while maintaining downstream signaling, and variant designs are centered on substitutions including K74A, E75Y, and N83R.

Functional bias is reported in terms of differential immune pathway activation, including reduced PD-L1 upregulation while substantially retaining or boosting MHC class I (HLA-ABC) expression. The disclosure further includes engineered architectures such as linked polypeptide segments, including cleavable peptide linker configurations, and provides example variants showing IFN-b3R2:IFN-b3R1 binding-affinity ratios alongside downstream signaling readouts including phospho-STAT1 and MHC I:PD-L1 expression ratio.

Claims Coverage

The provided independent claims cover IFN-b3 agonist recombinant polypeptides defined by high sequence identity to SEQ ID NO: 1 and specific amino-acid substitutions, including linked first/second amino acid sequences. The coverage further includes functional constraints such as receptor binding affinity ratio and immune-bias outcomes including reduced PD-L1 upregulation with retained MHC class I upregulation.

Across the independent claim set, coverage centers on IFN-b3 agonist recombinant polypeptides maintaining at least 95% identity to SEQ ID NO: 1 while incorporating specific substitutions at positions corresponding to K74, E75, and N83, with K74A, E75Y, and N83R explicitly specified in one independent claim. The disclosed claim landscape further includes operable linkage, linker configurations, and functional constraints involving receptor-binding affinity ratios and immune-bias outcomes.

Stated Advantages

Documented Applications