Antibody-drug conjugates through specific linker oligopeptides
Inventors
Spycher, Philipp • PROBST, Philipp • Attinger-Toller, Isabella • Bertrand, Romain • STARK, Ramona • Grabulovski, Dragan
Assignees
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Publication Number
US-12076412-B2
Publication Date
2024-09-03
Expiration Date
Abstract
The present invention relates to a method for generating an antibody-payload conjugate by means of a microbial transglutaminase (MTG). The method comprises a step of conjugating a linker comprising the structure (shown in N→C direction) (Sp1)-RK-(Sp2)-B-(Sp3) or (Sp1)-B-(Sp2)-RK-(Sp3) to a Gln residue comprised in an antibody, wherein (Sp1) is a chemical spacer or is absent; (Sp2) is a chemical spacer or is absent; (Sp3) is a chemical spacer or is absent; R is arginine or an arginine derivative or an arginine mimetic; K is lysine or a lysine derivative or a lysine mimetic; B is a linking moiety or a payload; and wherein the linker is conjugated to the Gin residue comprised in the antibody via a primary amine comprised in the side chain of the lysine residue, the lysine derivative or the lysine mimetic. Further, the invention relates to antibody-linker conjugates, antibody-drug conjugates and linker constructs comprising an RK motif.
Core Innovation
The invention relates to antibody-drug conjugates comprising an IgG antibody and a linker comprising a drug moiety B, wherein the drug moiety B is a toxin covalently linked to a peptide selected from RKAA, RKA, ARK, or RK-Val-Cit. The linker is conjugated to the IgG antibody via an isopeptide bond formed between the γ-carboxamide group of glutamine residue Q295 (EU numbering) of the CH2 domain of the antibody and the primary amine comprised in the side chain of a lysine residue comprised in the peptide.
The disclosed linker includes RK motif linker constructs and MTG-mediated site-specific antibody conjugation using linker peptides in the form (Sp1)-RK-(Sp2)-B-(Sp3) or (Sp1)-B-(Sp2)-RK-(Sp3). The document further describes linker variants including specific RK sequences, self-immolative spacers such as PABC, and cathepsin-cleavable motifs including Val-Cit, together with ether, thioether, and carbamate variants as part of the conjugation framework.
The invention states that conserved conjugation at IgG Q295 enables efficient coupling to glycosylated IgG while retaining the N297 glycan, and that this supports off-the-shelf production of IgG ADCs of basically any type. It also describes combining two different toxin or payload types to provide potential benefits such as increased cytotoxicity and targeting different cellular mechanisms, and it includes therapeutic and diagnostic use assertions.
Claims Coverage
The consolidated independent claim coverage centers on site-specific isopeptide bond conjugation between Q295 (EU numbering) in the IgG CH2 domain and a lysine side-chain primary amine in an RK peptide bearing a toxin drug moiety B. Across the independent claims, the coverage includes multiple specified antibody embodiments and peptide-linked toxin payload selections.
Isopeptide bond conjugation at Q295 (EU numbering)
The linker is conjugated to the IgG antibody via an isopeptide bond formed between the γ-carboxamide group of glutamine residue Q295 (EU numbering) of the CH2 domain and the primary amine comprised in the side chain of the lysine residue comprised in the peptide.
Toxin drug moiety B covalently linked to RK motif peptide
The drug moiety B is a toxin covalently linked to a peptide selected from RKAA, RKA, ARK, or RK-Val-Cit.
Antibody embodiments with specified IgG sequences
The IgG antibody is Polatuzumab or an antibody comprising a heavy chain as set forth in SEQ ID NO:5 and a light chain as set forth in SEQ ID NO:6, Trastuzumab or an antibody comprising a heavy chain as set forth in SEQ ID NO:7 and a light chain as set forth in SEQ ID NO:8, or Enfortumab or an antibody comprising a heavy chain as set forth in SEQ ID NO:9 and a light chain as set forth in SEQ ID NO:10 or 11.
Overall, the independent claims cover ADCs in which a toxin drug moiety B is covalently linked to an RK motif peptide and attached to an IgG antibody through a site-specific isopeptide bond between Q295 in the CH2 domain and a lysine side-chain primary amine in the linker. The claims further narrow the IgG antibody to Polatuzumab, Trastuzumab, Enfortumab, or specified heavy and light chain sequence sets.
Stated Advantages
Conserved conjugation site Q295 enables off-the-shelf production of IgG ADCs of basically any type.
Efficient conjugation for glycosylated antibodies while retaining the N297 glycan.
Efficient conjugation even for bulky payloads.
High conjugation efficiencies are reported as ≥82%, including examples described as up to ~100%.
The approach avoids deglycosylation or deleterious N297 substitution.
Potential benefits of combining two different toxin or payload types, including increased cytotoxicity and targeting different cellular mechanisms.
Documented Applications
Therapeutic use in neoplastic diseases is described.
Diagnostic use assertions, including radionuclide and near-infrared fluorescent dye combinations (SPECT/PET; fluorescence).
Example antibody targets such as Polatuzumab, Trastuzumab, and Enfortumab in connection with therapeutic use.
Nectin-4 and HER2/CD79b use cases as described.
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