Design of binder proteins specific to IL-13

Interleukin-13 (IL-13) drives inflammatory responses in type 2 inflammatory disorders by binding to the IL-4Rα/IL-13Rα1 receptor complex. Anti-IL-13 monoclonal antibodies such as tralokinumab and lebrikizumab showed substantial clinical efficacy in autoimmunology diseases such as atopic dermatitis and asthma. However, their large size has limited the possible application in topical immunotherapy treatment. Here, we employed structure-based computational design to develop compact minibinder proteins targeting IL-13, based on the crystal structure of the IL-13-tralokinumab complex. Five designed constructs were successfully expressed in E. coli and biophysically characterized. The purified minibinders displayed a range of binding affinities for IL-13, with binder1 showing the strongest interaction of nanomolar affinity. Functional evaluation in TF-1 cells showed that all minibinders inhibited IL-13-dependent proliferation in a concentration-dependent manner, with binder1 exhibiting the most potent antagonism (IC₅₀ = 56.39 nM). Furthermore, binder1 effectively suppressed STAT6 phosphorylation, and significantly attenuated IL-13-induced expression of pro-inflammatory genes in A549 lung epithelial cells. This study demonstrates the design of minimalistic, potent IL-13-targeting minibinders with potential application in topical immunotherapy treatment of IL-13-related inflammatory disorders.