# Protein engineered inhibitors of TNF Receptor activation: new therapeutics and new technology

> **NIH NIH R21** · UNIVERSITY OF MINNESOTA · 2020 · $176,342

## Abstract

ABSTRACT
Tumor necrosis factor (TNF) ligands and TNF receptors (TNFRs) are essential regulators of the immune
response. Dysregulation of TNF plays a role in the pathology of many autoimmune diseases that currently
afflict more than 23.5 million people in the U.S. Therapeutic targeting of TNFR1 signaling (e.g. for rheumatoid
arthritis, psoriasis, and inflammatory bowel disease) is a billion-dollar industry. However, the available anti-
TNF agents cause severe and adverse side effects. Thus, there is a desperate need to develop 'anti-TNFR'
instead of 'anti-TNF' treatments in chronic inflammatory and autoimmune disorders. Despite some recent
progress in this regard, state-of-the-art small molecule approaches have failed to uncover any high affinity
small molecule inhibitors.
In an attempt to jumpstart renewed and needed therapeutic discovery efforts, we have been building on
existing yeast display/directed evolution technology to engineer high affinity TNFR ligands, in place of small
molecules. Protein ligand scaffolds, peptides with high affinity and large surface area, are engineered by
modulating amino acids in a select region, known as the paratope, of a protein while conserving a stable
underlying scaffold. One particular example, the affibody domain, which has been extensively studied and
improved by co-Investigator Hackel, has been effectively used as a ligand scaffold to numerous targets, with
affinities as strong as 20 pM, and application to diagnostics, molecular imaging, and therapy.
However, as we progressed towards high affinity binders to the TNFR family, we reached a familiar bottleneck
in the field: how to direct the evolution of binders based not on affinity, but on functionality. While numerous
platforms exist for discovery and evolution of protein binding, no robust methods have been established for the
selection of precise biological activity (aside from general survival screens).
Thus, the objective of this proposal is the development of a new technology for activity-based, high-throughput
screening of protein ligands. In so-doing, we will discover novel, high-affinity inhibitors of TNFRs. Aim 1 will
achieve discovery and evolution of a broad panel of strong binders to TNFRs, though the frequency of
functional inhibitors is expected to be quite low. Aim 2 develops a technology to dramatically enhance the
discovery of functional binders, which will have broad utility for all active ligand screening in addition to a
focused benefit on the current TNFR antagonist development.

## Key facts

- **NIH application ID:** 9859335
- **Project number:** 5R21AI144932-02
- **Recipient organization:** UNIVERSITY OF MINNESOTA
- **Principal Investigator:** Jonathan N Sachs
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $176,342
- **Award type:** 5
- **Project period:** 2019-02-01 → 2021-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9859335

## Citation

> US National Institutes of Health, RePORTER application 9859335, Protein engineered inhibitors of TNF Receptor activation: new therapeutics and new technology (5R21AI144932-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/9859335. Licensed CC0.

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