Polypeptides and uses thereof
Inventors
Grinstaff, Mark • HAMOUD, Aladin • Berry, Samantha • GROMISCH, Christopher • Ringaci, Alina
Assignees
Publication Number
US-12281161-B2
Publication Date
2025-04-22
Expiration Date
2042-07-26
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Abstract
Provided herein are novel polypeptides, methods for their production and uses thereof.
Core Innovation
The invention provides antibodies in which a polypeptide chain, such as the heavy chain, is linked at its C-terminus to a coiled-coil peptide capable of forming a heterodimer with another coiled-coil peptide. Specific amino acid sequences for these coiled-coil peptides are defined, including but not limited to SEQ ID NOs: 1, 5–8, 10–13. The coiled-coil peptide can be directly linked to the antibody or connected via a peptide linker, such as those recognized by Sortase A (e.g., LPETG, SEQ ID NO: 48), to facilitate selective and efficient attachment.
The background identifies antibody-drug conjugates (ADCs) as an area where improvements are sought, particularly in the stability of linkers and expansion of small molecule drug options. The presented technology addresses these needs by employing coiled-coil peptides to mediate site-specific and orthogonal attachment of various payloads, such as drugs, fluorophores, or other agents, to antibodies. The coiled-coil peptides offer a flexible platform, enabling selective heterodimeric assembly through predefined sequences.
A key aspect involves a two-component system: (i) an antibody chain (typically the heavy chain) with a coiled-coil peptide linked at the C-terminus, and (ii) a second coiled-coil peptide conjugated to a payload, such as a therapeutic, imaging, or detectable agent. The pairwise-specific coiled-coil interaction brings the payload into stable association with the antibody, enabling targeted delivery. This system can employ payload linkers with properties such as enzymatic cleavability (e.g., EVC tripeptide) and self-immolative moieties (e.g., PABC), enhancing control over payload release profiles.
Claims Coverage
There are two independent claims that define major inventive features relating to antibodies with engineered coiled-coil peptide linkages and their use in conjugate payload delivery.
Antibody with polypeptide chain linked at its C-terminus to a coiled-coil peptide
The invention features an antibody in which a polypeptide chain is linked at the C-terminus to a coiled-coil peptide, where: - The coiled-coil peptide is designed to form a heterodimer with another coiled-coil peptide. - Specific sequences for the coiled-coil peptides are recited (including SEQ ID NOs: 1, 5, 6, 7, 8, 10, 11, 12, 13). - The antibody comprises a second coiled-coil peptide (not linked to the antibody), able to form a heterodimer with the antibody-linked coiled-coil and further comprising a ligand or payload. - The second coiled-coil peptide includes specific sequences (e.g., SPEDEIQALEEENAQLEQENAALEEELAQLEYGK-azidoG, SEQ ID NO: 4; or analogous sequences with lysine-modification for payload conjugation, SEQ ID NOs: 51 and 52). - The ligand may be a therapeutic, imaging agent, detectable label, or other compound, and can be attached via cleavable linkers (such as EVC or PEG3-EVC-PABC).
Antibody with heavy chain linked to a coiled-coil peptide, wherein the conjugated ligand is covalently linked with a coiled-coil peptide
This inventive feature covers: - An antibody where the heavy chain is linked at its C-terminus to a coiled-coil peptide, specifically SEQ ID NO: 10. - The coiled-coil is attached via a peptide linker containing the sequence GGGGG (SEQ ID NO: 55). - The antibody is conjugated with a ligand through a second coiled-coil peptide, which incorporates a sequence for selective interaction with the antibody-linked peptide (e.g., SEQ ID NO: 52). - The ligand/payload is covalently attached to the coiled-coil peptide, thus forming a modular antibody–payload conjugate through heterodimeric coiled-coil pairing.
The claims encompass antibodies with precisely engineered C-terminal coiled-coil peptides enabling heterodimeric assembly with payload-bearing coiled-coil peptides, where payloads are attached through cleavable linkers to achieve targeted delivery and modular construction.
Stated Advantages
Provides orthogonal, modular, and site-specific conjugation of diverse payloads to antibodies using coiled-coil peptide interactions, allowing flexible configuration of antibody–payload systems.
Permits use of cleavable linkers, such as enzyme-sensitive or self-immolative linkers, enhancing control over payload release at target sites.
Enables straightforward and efficient method for preparing antibody conjugates without affecting antibody binding affinity or pharmacokinetic properties, as described in plasma stability and in vivo clearance characterization.
Minimizes the need for harsh chemical modifications or random conjugation, reducing risk of compromising antibody structure or function.
Documented Applications
Targeted delivery of therapeutic agents, such as anti-cancer drugs, to disease-relevant cells using antibody-drug conjugates assembled via coiled-coil peptides.
Use of antibody–coiled-coil conjugates for delivery of imaging agents or detectable labels to cells for diagnostic purposes.
Preparation and administration of pharmaceutical compositions comprising antibody–payload conjugates for treating diseases or disorders, including various forms of cancer.
Delivery of ligands, including nucleic acids, peptides, oligonucleotides, or other biologically active molecules, to target cells in research or therapeutic applications.
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