Activatable Antibody Conjugation via Partial Disulfide Reduction
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Solution Overview
Problem
Existing antibody-based therapeutics face limitations such as rapid clearance from circulation and toxicities due to broad target expression, and there is a need for improved methods of conjugating agents to antibodies without compromising their therapeutic effectiveness.
Innovation Solution
The development of activatable antibodies with masking and cleavable moieties that allow for selective conjugation of agents at specific sites without disturbing disulfide bonds, maintaining the antibody's binding and masking activities.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If agents are conjugated to antibodies using conventional methods, then the therapeutic and diagnostic functionality is enhanced, but the antibody's binding activity and masking capabilities are compromised
Solution Approach 1:
The patent segments the antibody structure by introducing a separate masking moiety (MM) that can be independently controlled from the antibody binding region. This allows the antibody (AB) to maintain its binding capability while the MM provides selective masking, and conjugation can be performed on the AB without affecting the MM's masking function.
Solution Approach 2:
The patent uses a cleavable moiety (CM) as an intermediary element that links the masking moiety to the antibody. The CM is designed to be cleaved by specific proteases, allowing the MM to be released from the AB only when needed. This intermediary enables controlled interaction between components while preserving their individual functionalities.
2Productivity
If conventional conjugation methods are used, then agents can be attached to antibodies, but rapid clearance from circulation occurs and toxicities increase
Solution Approach 1:
The patent applies local quality by creating a specific conjugation site on the antibody that is distinct from the binding site and masking site. The antibody is engineered with a specific cysteine residue or reduced disulfide bond location that allows agent attachment without interfering with the binding or masking functions, thereby reducing off-target effects and clearance.
Solution Approach 2:
The patent performs preliminary reduction of specific disulfide bonds in the antibody to create designated conjugation sites before agent attachment. This preliminary action ensures that conjugation occurs at predetermined locations that do not interfere with binding or masking, preventing subsequent loss of function and reducing clearance.
3Adaptability or versatility
If all disulfide bonds are reduced for conjugation, then maximum conjugation sites are available, but the masking and binding activities are lost
Solution Approach 1:
The patent applies partial action by selectively reducing only specific disulfide bonds in the antibody rather than all disulfide bonds. This partial reduction creates sufficient conjugation sites for agent attachment while preserving other disulfide bonds that maintain the antibody's binding and masking activities.
Solution Approach 2:
The patent creates local quality differences within the antibody structure by reducing only specific disulfide bonds located in regions designated for conjugation, while leaving other disulfide bonds intact in regions critical for binding and masking functions. This spatial differentiation allows simultaneous achievement of conjugation capability and functional preservation.
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 enables the creation of conjugated activatable antibodies that retain their masking and binding capabilities, allowing for targeted therapy and diagnosis while minimizing adverse effects.
Implementation Method 1
partially reducing at least one interchain disulfide bond in the activatable antibody with a reducing agent to form at least two interchain thiols
Implementation Method 2
conjugating the agent to the reduced activatable antibody
Data Source
AI summary
The invention relates generally to compositions and methods for conjugating antibodies and activatable antibodies, and methods of partially reducing antibodies and/or activatable antibodies prior to conjugation, e.g., thiol-based conjugation, with an agent, e.g., a therapeutic and/or diagnostic agent.