BRISC Inhibitor Design via Allosteric Dimerization

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current methods lack selective inhibitors for the BRISC DUB complex due to cross-reactivity with other metalloenzymes, and obtaining high-resolution structures for inhibitor design is challenging due to conformational flexibility and complex dissociation.

Innovation Solution

An integrative structural biology approach using cryo-electron microscopy to identify unique modes of inhibition, revealing a new BRISC conformer and binding site, and employing BRISC dimers to facilitate inhibitor design through allosteric binding and detection methods like native mass spectrometry and FRET/BRET pairs.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If broad-spectrum zinc chelators are used to inhibit BRISC, then BRISC DUB activity is suppressed, but cross-reactivity with other metalloenzymes occurs

Engineering Contradiction:

Improveinhibitor specificityVSAvoidcross-reactivity with other metalloenzymes

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent applies local quality by targeting a specific allosteric binding site on the BRISC complex rather than using non-specific zinc chelators. The inhibitor binds to a unique pocket formed by the U-shaped arrangement of BRCC36, Abraxas2, BRCC45, and MERIT40 subunits, creating a locally specific interaction that does not cross-react with other metalloenzymes while maintaining selective BRISC inhibition.

Inventive Principle:

Principle #3Local quality

Solution Approach 2:

The patent uses an intermediary approach by employing a molecular glue compound that mediates between the BRISC subunits. This compound binds allosterically and stabilizes a dimeric conformation of the BRISC complex, acting as a mediator that induces structural changes without directly inhibiting the catalytic zinc site, thereby avoiding cross-reactivity issues.

Inventive Principle:

Principle #24Intermediary (Mediator)

2Measurement precision

If cryo-electron microscopy is used to obtain BRISC structures, then high-resolution structures can be obtained, but conformational flexibility and complex dissociation prevent successful crystallization

Engineering Contradiction:

Improvestructural resolutionVSAvoidcomplex stability

Core Design Contradiction:

Measurement precisionVSStability of the object's composition

Solution Approach 1:

The patent applies preliminary action by pre-stabilizing the BRISC complex in a dimeric conformation before structural analysis. The allosteric inhibitor is added to the BRISC complex prior to cryo-EM sample preparation, locking the complex into a stable dimeric state that resists dissociation during the cryo-EM process, thereby enabling successful high-resolution structure determination.

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent utilizes parameter changes by inducing a conformational transition in the BRISC complex from a flexible monomeric or dissociated state to a stable dimeric state. This structural parameter change (from dispersed to associated conformations) enhances the stability of the complex for cryo-EM analysis while maintaining high-resolution structural information.

Inventive Principle:

Principle #35Parameter changes

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This approach allows for the identification of selective BRISC inhibitors that act as 'molecular glues' to form dimeric complexes, sterically blocking the active site and enhancing inhibitor specificity, and provides stable protein samples for structural studies.

Implementation Method 1

employing BRISC dimers to facilitate inhibitor design through allosteric binding

Methodology Applied

Scientific EffectAllosteric binding:

Implementation Method 2

identification of selective BRISC inhibitors that act as 'molecular glues' to form dimeric complexes

Methodology Applied

Scientific EffectDimerization:

Implementation Method 3

sterically blocking the active site and enhancing inhibitor specificity

Methodology Applied

Scientific EffectSteric blocking:

Implementation Method 4

detection methods like native mass spectrometry and FRET/BRET pairs

Methodology Applied

Scientific EffectMass spectrometry:

Implementation Method 5

detection methods like native mass spectrometry and FRET/BRET pairs

Methodology Applied

Scientific EffectFluorescence resonance energy transfer: Fluorescence

Implementation Method 6

detection methods like native mass spectrometry and FRET/BRET pairs

Methodology Applied

Scientific EffectBioluminescence resonance energy transfer: Bioluminescence

Implementation Method 7

An integrative structural biology approach using cryo-electron microscopy to identify unique modes of inhibition

Methodology Applied

Scientific EffectCryo-electron microscopy:

Data Source

PatentUS20230317203A1Screening method

Publication Date: 2023.10.05 UNIVERSITY OF LEEDS

AI summary

The present invention relates to method of identifying a selective BRISC inhibitor. The present invention also relates to a stable BRISC dimer. The present invention further relates to use of the stable BRISC dimer to generate cryo-Electron Microscopy (cryo-EM), crystallography, nuclear magnetic resonance and/or X-ray crystallography structures for structure guided drug design.