Non-IL-2 Blocking Anti-CD25 Antibody for Treg Depletion

Resolve Bottlenecks,
Find Innovative Solutions
Generate Solutions

Solution Overview

Problem

Current cancer immunotherapies face limitations due to the infiltration of regulatory T cells (Tregs) in tumors, which suppress anti-tumor immune responses by altering the Teff/Treg ratio, necessitating a targeted approach to deplete Tregs without inhibiting IL-2 signaling to enhance anti-tumor immunity.

Innovation Solution

Development of a non-IL-2 blocking anti-CD25 antibody through hybridoma screening and humanization, which selectively binds to human CD25, depletes Tregs while allowing IL-2 to stimulate effector T cells, thereby correcting the tumor microenvironment.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If existing anti-CD25 antibodies (Basiliximab, Daclizumab) are used to deplete Tregs, then Treg depletion is achieved, but IL-2 signaling is inhibited which reduces anti-tumor immunity

Engineering Contradiction:
ImproveTreg depletion efficacyVSAvoidIL-2 signaling inhibition
Core Design Contradiction:
ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The antibody molecule is divided into distinct functional regions: the variable region (VH-VL) provides high-affinity binding to CD25 for Treg depletion, while the constant region (CH2-CH3) is engineered to avoid IL-2 receptor blockade. This segmentation allows independent optimization of Treg depletion capability while preserving IL-2 signaling pathways.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The antibody exhibits localized functionality: the antigen-binding site specifically targets CD25 on Tregs with high affinity, while the Fc region is designed to mediate ADCC without interfering with IL-2 binding to its natural receptor. This spatial separation of functions enables selective Treg depletion while maintaining anti-tumor immune responses.

Inventive Principle:
Principle #3Local quality

2Reliability

If Tregs are depleted to increase Teff/Treg ratio, then anti-tumor immunity is enhanced, but existing antibodies cause off-target effects by blocking IL-2 signaling

Engineering Contradiction:
ImproveTeff/Treg ratio improvementVSAvoidOff-target IL-2 receptor blockade
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The antibody serves as a selective intermediary that binds CD25 on Tregs to trigger ADCC-mediated depletion, while its engineered structure prevents interference with IL-2 binding to the complete IL-2 receptor complex on Teff cells. This intermediary action achieves Treg depletion without the off-target effects of previous antibodies.

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The antibody's binding parameters were optimized through humanization and affinity maturation to achieve high specificity for CD25 on activated Tregs. The binding affinity (Kd) and specificity were tuned to distinguish between Treg CD25 and the IL-2 receptor complex, enabling selective depletion without blocking IL-2 signaling to effector cells.

Inventive Principle:
Principle #35Parameter changes

3Reliability

If non-IL-2 blocking anti-CD25 antibody is developed, then both Treg depletion and IL-2 signaling preservation are achieved, but development complexity increases

Engineering Contradiction:
ImproveDual functionality (Treg depletion + IL-2 signaling preservation)VSAvoidAntibody development process
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The antibody development process incorporated preliminary humanization of the murine antibody framework, which pre-positioned the variable regions to bind CD25 with high affinity while using human constant regions that are incompatible with blocking IL-2 signaling. This preliminary structural preparation simplified subsequent optimization steps and ensured the desired dual functionality from the outset.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The successful antibody structure was copied and validated through multiple sequence variants (SEQ ID NO: 1-25 for VH, SEQ ID NO: 26-62 for VL) that maintained the core functional architecture. By copying and refining the proven design, the complex functionality was achieved through systematic optimization rather than de novo design.

Inventive Principle:
Principle #26Copying

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 antibody effectively increases the Teff/Treg ratio, enhancing the killing ability of effector T cells and improving anti-tumor responses by maintaining IL-2 signaling, thus offering a significant clinical benefit in cancer treatment.

Implementation Method 1

A CD25-targeted antibody with antibody-dependent cell-mediated cytotoxicity (ADCC) effect can effectively deplete Tregs in a tumor microenvironment

Methodology Applied
Scientific EffectAntibody-dependent cell-mediated cytotoxicity (ADCC):

Implementation Method 2

IL-2 is a key cytokine for Teff responses. Therefore, antibodies targeting CD25 on the one hand can effectively deplete Tregs in the tumor microenvironment and on the other hand do not inhibit the IL-2 signaling through CD25, so that not only Treg cells can be removed mechanically, but also IL-2 can be allowed to stimulate Teff cells

Methodology Applied
Scientific EffectCytokine signaling:

Data Source

PatentEP4386000A1Recombinant Anti-human-CD25 antibody and use thereof
Publication Date: 2024.06.19 NANJING NOVOACINE BIO-TECH CO LTD
  • EP4386000A1 patent drawingFigure 1A~1B
  • EP4386000A1 patent drawingFigure 2A~2D
  • EP4386000A1 patent drawingFigure 3~4

AI summary

An anti-human-CD25 antibody molecule or an antigen-binding fragment thereof. Compared with an existing anti-CD25 antibody, the provided antibody is non-IL-2 inhibitory, and can not only eliminate Treg, but can also correct a tumor microenvironment and increase the proportion and killing ability of Teff. Further provided is the use of the antibody molecule or the antigen-binding fragment in the preparation of a drug for treating solid tumors and hematological tumors.