PD-1 Receptor Mutants for Selective PD-L1 Over PD-L2 Binding

Solution Overview

Problem

Existing therapeutic agents, such as monoclonal antibodies, have limitations in targeting the PD-1 signaling pathway due to their large size and antigenicity, and there is a need for PD-1 receptor mutants with selective binding to PD-L1 over PD-L2 for improved immune response modulation and imaging.

Innovation Solution

Computational redesign of the PD-1 receptor using ProtLID to generate residue-based pharmacophores for PD-L1 and PD-L2 interfaces, followed by site-directed mutagenesis to create mutants with specific mutations like N66Q, Y68N, Y68R, Y68Q, S73R, Q75N, T76D, T76E, D77E, K78R, G124V, L128V, K131H, I134N, or I134F, enhancing affinity to PD-L1 while reducing or eliminating binding to PD-L2.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If monoclonal antibodies are used to block PD-1 signaling pathway, then therapeutic activity against cancers is achieved, but antigenicity and poor tissue penetrance occur due to large size

Engineering Contradiction:

Improvetherapeutic activityVSAvoidantigenicity and poor tissue penetrance

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The invention extracts the essential functional domain of PD-1 (the ectodomain containing residues 25-168) and uses it to create simplified receptor mutants. This extraction eliminates the harmful properties of full-sized monoclonal antibodies (large size, antigenicity) while retaining the therapeutic function of blocking PD-1 signaling pathway and enhancing anti-tumor immune responses

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The invention applies parameter changes by introducing specific point mutations (N66Q, Y68N, Y68R, Y68Q, S73R, Q75N, T76D, T76E, D77E, K78R, G124V, L128V, K131H, I134N, or I134F) into the PD-1 ectodomain to create mutants with selective binding to PD-L1. These parameter changes at the molecular level achieve the desired selectivity without the drawbacks of large antibody molecules

Inventive Principle:

Principle #35Parameter changes

2Force

If PD-1 receptor mutants with multiple mutations are engineered to achieve high affinity to PD-L1, then binding affinity increases 15-40 thousand fold, but antigenic properties deteriorate

Engineering Contradiction:

Improvebinding affinityVSAvoidantigenic properties

Core Design Contradiction:

ForceVSObject-affected harmful factors

Solution Approach 1:

The invention applies parameter changes by introducing specific point mutations (N66Q, Y68N, Y68R, Y68Q, S73R, Q75N, T76D, T76E, D77E, K78R, G124V, L128V, K131H, I134N, or I134F) into the PD-1 ectodomain to create mutants with selective binding to PD-L1. These parameter changes at the molecular level achieve the desired selectivity without the drawbacks of large antibody molecules

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The invention applies local quality by making targeted mutations only at specific residues (26 out of 168 residues) within the PD-1 ectodomain that are critical for PD-L1 binding. This localized modification approach achieves high affinity and selectivity while minimizing overall structural changes and potential antigenic properties

Inventive Principle:

Principle #3Local quality

3Measurement precision

If PD-1 receptor mutants are designed to bind selectively to PD-L1, then imaging and treatment of PD-L1 positive tumors is improved, but binding selectivity must be optimized to eliminate cross-reactivity with PD-L2

Engineering Contradiction:

Improvebinding selectivityVSAvoidcross-reactivity with PD-L2

Core Design Contradiction:

Measurement precisionVSObject-affected harmful factors

Solution Approach 1:

The invention applies local quality by making targeted mutations only at specific residues (26 out of 168 residues) within the PD-1 ectodomain that are critical for PD-L1 binding. This localized modification approach achieves high affinity and selectivity while minimizing overall structural changes and potential antigenic properties

Inventive Principle:

Principle #3Local quality

Solution Approach 2:

The invention applies parameter changes by introducing specific point mutations (N66Q, Y68N, Y68R, Y68Q, S73R, Q75N, T76D, T76E, D77E, K78R, G124V, L128V, K131H, I134N, or I134F) into the PD-1 ectodomain to create mutants with selective binding to PD-L1. These parameter changes at the molecular level achieve the desired selectivity without the drawbacks of large antibody molecules

Inventive Principle:

Principle #35Parameter changes

Data Source

PatentUS12390541B2Mutant variants of PD-1 receptor with selective binding to PD-L1 and uses thereof

Publication Date: 2025.08.19 ALBERT EINSTEIN COLLEGE OF MEDICINE OF YESHIVA UNIV

AI summary

Disclosed are human Programmed Cell Death-1 (PD-1) receptor mutants having selectivity for PD-L1 compared to PD-L2, methods of obtaining the mutants, and uses of the mutants for treatment and imaging.