Humanized CD19 CAR-T Cells with CDR1 V27G Mutation

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Solution Overview

Problem

Current adoptive T cell immunotherapy for cancer has limitations in terms of efficacy and safety, with a need for improved treatments that can effectively target cancer cells while minimizing toxicity.

Innovation Solution

Development of humanized CD19-CAR-T cells with a mutation in the CD19 ScFv VH CDR1 region (clone 11), which includes a chimeric antigen receptor (CAR) fusion protein comprising a humanized single-chain variable fragment (scFv) of CD19, a transmembrane domain, at least one co-stimulatory domain, and an activating domain.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional CAR-T cells are used for immunotherapy, then tumor targeting capability is achieved, but efficacy and safety are limited due to insufficient specificity and high toxicity

Engineering Contradiction:

Improveefficacy and safety of immunotherapyVSAvoidtoxicity

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent applies local quality by introducing a specific mutation (V27G) in the CDR1 region of the VH domain of the CD19 scFv antibody. This localized modification enhances the binding specificity to human CD19 antigen while minimizing off-target effects, thereby improving therapeutic efficacy and reducing toxicity simultaneously

Inventive Principle:

Principle #3Local quality

Solution Approach 2:

The patent changes the amino acid parameter at position 27 of the VH CDR1 region from Valine to Glycine. This parameter change optimizes the antigen-binding properties of the CAR-T cells, enabling them to specifically recognize and target human CD19+ leukemia cells with higher precision and reduced harmful effects

Inventive Principle:

Principle #35Parameter changes

2Measurement precision

If humanized CD19-CAR-T cells with CDR1 mutation are developed, then specificity to leukemia cells is enhanced, but device complexity increases

Engineering Contradiction:

Improvespecificity of targetingVSAvoidcomplexity of CAR structure

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The patent segments the CAR structure into distinct functional domains: a humanized CD19 scFv with V27G mutation for antigen recognition, a hinge region for flexibility, a transmembrane domain for cell membrane integration, and intracellular signaling domains (CD28 and CD3ζ) for T cell activation. This segmentation allows each domain to be optimized independently while maintaining overall functionality

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent creates a composite CAR structure by combining humanized antibody fragments (scFv with V27G mutation) with T cell signaling components (CD28 co-stimulatory domain and CD3ζ activation domain). This composite design integrates multiple functional elements into a single receptor that achieves high specificity while maintaining manageable structural complexity through modular organization

Inventive Principle:

Principle #40Composite materials

Data Source

PatentUS12202896B2CAR-T cells with humanized CD19 ScFv with mutation in CDR 1 region

Publication Date: 2025.01.21 ZHEJIANG RUIJIAMEI BIOTECH CO LTD

AI summary

Provided is a humanized CD19 single-chain variable fragment (scFv) having a mutation of valine to glycine in CDR1, comprising VH having the amino acid sequence of SEQ ID NO: 6 and VL having the amino acid sequence of SEQ ID NO: 5. Also provided is a CD19 chimeric antigen receptor fusion protein comprising from N-terminus to C-terminus: (i) a single-chain variable fragment (scFv), (ii) a transmembrane domain, (iii) at least one co-stimulatory domains, and (iv) an activating domain. This humanized CD19-CAR-T cells have specific killing activity with secretion of cytokine IFN-gamma in CAR-T cells in vitro and in vivo.