Robo Receptor Inhibitors Targeting Dimerization Interfaces

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current approaches to inhibit Robo receptor activity, such as using soluble fragments or antibodies, face challenges due to the high affinity of Slit-Robo interactions and local concentration of Slit, which diminish their effectiveness in vivo.

Innovation Solution

Development of an agent that specifically binds to Robo receptor ectodomains, particularly the Ig-like domain 3, Ig-like domain 4, Fibronectin type III domains, and their hinge regions, to inhibit dimerization, thereby blocking Robo receptor activation without binding to Slit-binding domains like D1 or D2.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If soluble fragments or antibodies are used to inhibit Robo receptor activity, then Robo signaling can be blocked, but the high affinity of Slit-Robo interactions and local concentration of Slit diminish effectiveness in vivo

Engineering Contradiction:

Improveeffectiveness of Robo inhibitionVSAvoidhigh affinity Slit-Robo interaction

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The invention divides the Robo receptor ectodomain into specific functional segments (Ig-like domains D3-D8 and hinge regions) and targets each segment with specific inhibitory agents. This segmentation allows the agents to bind to specific dimerization interfaces (D3-D3 or D4-D4) without interfering with Slit binding to D1-D2, thereby resolving the contradiction between blocking Robo signaling and maintaining Slit-Robo interaction affinity.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The invention applies local quality by designing agents that specifically target particular local regions of the Robo receptor (D3-D8 domains and hinge regions) rather than binding to the Slit-binding sites (D1-D2). This localized binding enables selective inhibition of Robo dimerization and signaling while leaving the Slit-Robo interaction interface intact, thus maintaining high affinity binding where needed.

Inventive Principle:

Principle #3Local quality

2Reliability

If agents bind to Slit-binding domains (D1 or D2) to inhibit Robo activity, then Robo signaling is blocked, but the high local concentration of Slit reduces effectiveness

Engineering Contradiction:

Improveinhibition of Robo signalingVSAvoidlocal concentration of Slit

Core Design Contradiction:

ReliabilityVSQuantity of substance

Solution Approach 1:

The invention extracts the inhibitory function from the Slit-binding domains (D1-D2) and relocates it to the dimerization domains (D3-D8 and hinge regions). By taking out the binding target from the Slit-interaction zone and placing it in the dimerization zone, the agent can effectively inhibit Robo signaling without being outcompeted by high local concentrations of Slit, since the agent no longer competes for Slit binding sites.

Inventive Principle:

Principle #2Taking out (Extraction)

3Reliability

If non-specific binding to Robo receptor is used, then general inhibition is achieved, but specificity and selectivity are reduced

Engineering Contradiction:

Improveinhibition of Robo activityVSAvoidspecificity of binding

Core Design Contradiction:

ReliabilityVSMeasurement precision

Solution Approach 1:

The invention implements local quality by designing agents with specific binding sites for particular domains (D3-D8) and hinge regions of the Robo receptor. This localized specificity ensures that the agents selectively inhibit Robo dimerization at the correct interfaces without non-specific binding to other regions, thereby achieving both reliable inhibition and precise binding specificity.

Inventive Principle:

Principle #3Local quality

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

The agent effectively inhibits Robo receptor dimerization with a micromolar or sub-micromolar dissociation constant, specifically targeting the D4-D4 or D3-D3 interfaces, thereby disrupting intracellular signaling and reducing Robo receptor activity.

Implementation Method 1

an agent capable of specifically binding to a Robo receptor ectodomain polypeptide selected from the group consisting of a Robo receptor Immunoglobulin-like domain 3 (D3), a Robo receptor Immunoglobulin-like domain 4 (D4), a Robo receptor Immunoglobulin-like domain 5 (D5), a Robo receptor Fibronectin type III domain 1 (FnIII-1)(D6), a Robo receptor Fibronectin type III domain 2 (FnIII-2)(D7), a Robo receptor Fibronectin type III domain 3 (FnIII-3)(D8), a Robo receptor D3-D4 hinge region, a Robo receptor D4-D5 hinge region, a Robo receptor D5-D6 hinge region, a Robo receptor D6-D7 hinge region and a Robo receptor D7-D8 hinge region, or a portion thereof

Methodology Applied

Scientific EffectSpecific binding:

Implementation Method 2

inhibiting Robo receptor dimerization... specifically targeting the D4-D4 or D3-D3 interfaces, thereby disrupting intracellular signaling and reducing Robo receptor activity

Methodology Applied

Scientific EffectDimerization inhibition:

Data Source

PatentUS11406682B2Roundabout (Robo) receptor inhibitors and uses thereof

Publication Date: 2022.08.09 BAR ILAN UNIV

AI summary

The present invention, in some embodiments thereof, relates to inhibitory compositions binding to the ectodomain of a Roundabout (Robo) receptor and to the use of same for downregulating Robo-mediated signaling. In particular, the present invention provides moieties that bind to and prevent dimerization of an Ig-like Robo receptor ectodomain, thereby inhibiting Robo receptor activity.