# Engineering selective inhibition of metalloproteinases by tissue inhibitors of metalloproteinases (R01 GM132100 RESUB - *TIMPs)

> **NIH NIH R01** · MAYO CLINIC  JACKSONVILLE · 2022 · $313,000

## Abstract

PROJECT SUMMARY/ABSTRACT
Matrix metalloproteinases (MMPs) have long been regarded as promising therapeutic targets, but clinical trials
of early-generation MMP inhibitors in arthritis and cancer proved disappointing. Broad-spectrum MMP
inhibitors produced serious dose-limiting musculoskeletal toxicity and failed to extend progression-free survival
in cancer trials, partly due to the inability of the inhibitors to distinguish among different MMPs. This was a
critical problem, because some MMPs serve a primarily protective function, and it is now clear that
indiscriminate inhibition of all MMPs inevitably leads to poorer outcomes. Based on our published and
preliminary data, we hypothesize that tissue inhibitors of metalloproteinases (TIMPs), endogenous regulators
of the MMP family, can be engineered into highly selective MMP inhibitors free of undesired off-target
activities, to produce probes and therapeutics targeting individual MMPs with exquisite selectivity. In this
application, we will (a) optimize novel methodology for directed evolution of selective binders to discriminate
within the large families of related MMPs and adamalysin proteases, (b) uncover mechanisms of molecular
recognition that govern MMP-TIMP binding specificity, and (c) develop a toolbox of engineered TIMPs that
selectively target individual MMPs with highly enhanced specificity. To accomplish these goals, we will use
state-of-the-art directed evolution approaches to engineer the TIMP-1 scaffold for fine discrimination between
closely similar MMPs, reengineering N- and C-terminal TIMP domain epitopes and exploiting cooperativity
between domains. Additionally, we will integrate X-ray crystallographic and computational approaches to
elucidate the protein structural and dynamic features that govern affinity and selectivity of TIMP/MMP
complexes. This project will thus elucidate fundamental principles of molecular recognition governing
TIMP/MMP selectivity, and will produce designer TIMPs targeting single MMPs with highly enhanced
specificity, with potential for development as useful molecular probes and as protein therapeutics for the many
diseases driven by MMP dysregulation.

## Key facts

- **NIH application ID:** 10319170
- **Project number:** 5R01GM132100-03
- **Recipient organization:** MAYO CLINIC  JACKSONVILLE
- **Principal Investigator:** Evette S Radisky
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $313,000
- **Award type:** 5
- **Project period:** 2020-04-01 → 2023-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10319170

## Citation

> US National Institutes of Health, RePORTER application 10319170, Engineering selective inhibition of metalloproteinases by tissue inhibitors of metalloproteinases (R01 GM132100 RESUB - *TIMPs) (5R01GM132100-03). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10319170. Licensed CC0.

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