Method for in vivo generation of multispecific antibodies from monospecific antibodies
Inventors
Brinkmann, Ulrich • Mayer, Klaus • Dickopf, Steffen
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-12180279-B2
Publication Date
2024-12-31
Expiration Date
Abstract
Herein is reported a method for the generation of multispecific antibodies directly on the cell-surface at the site of action by a half-antibody exchange reaction between two 2/3-IgGs or two 2/3-BiFabs destabilized in one half by asymmetric perturbing mutations fostering the generation of correctly assembled full length bi- or multispecific antibodies. The method is performed in the absence hinge region disulfide bonds in the starting 2/3-IgGs or 2/3-BiFabs.
Core Innovation
The invention relates to multimeric polypeptides comprising a first polypeptide and a second polypeptide arranged in an N- to C-terminal direction. Each polypeptide includes an antibody variable domain selected from a pair of an antibody light and heavy chain variable domain and a human immunoglobulin G CH3 domain. The variable-domain pairs are configured to specifically bind a first target, a second target, and a third target, with the second target positioned either N-terminal to the first antibody variable domain or C-terminal to the first CH3 domain.
The second polypeptide includes a second antibody variable domain specifically binding to a third target and a second human immunoglobulin G CH3 domain, and lacks a pair of an antibody light and heavy chain variable domain specifically binding to a target. The second CH3 domain comprises a perturbing mutation selected from D356K, E357K, K370E, and K439E, with corresponding residue requirements in the first CH3 domain, and all numbering is according to Kabat EU index.
The document further describes half-antibody exchange and on-cell in vivo assembly using destabilized, non-complete antibody formats, including 2/3-IgGs or 2/3-BiFabs, lacking inter-heavy-chain disulfide bonds. It also recites higher-order constructs and associated binding-site design, including heterotrimeric constructs, multimeric IgG formats, disulfide-linked third polypeptides, and pharmaceutical formulations containing the described entities.
Claims Coverage
The partial document contains two independent claims. Both independent claims define multimeric polypeptides built from two polypeptides containing human IgG CH3 domains and antibody light/heavy variable-domain pairs, and both require a specific CH3 perturbing-mutation scheme with Kabat EU numbering; the claims differ mainly in the order of the CH3 and variable domains on the first versus second polypeptides.
Multimeric polypeptide with first and second polypeptides and CH3 perturbing mutation scheme
A multimeric polypeptide comprising a first polypeptide including a first antibody variable domain specifically binding to a first target and a first human immunoglobulin G CH3 domain, and including a pair of antibody light and heavy chain variable domains specifically binding to a second target either N-terminal to the first antibody variable domain or C-terminal to the first CH3 domain; and a second polypeptide including a second antibody variable domain specifically binding to a third target and a second human immunoglobulin G CH3 domain, wherein the second polypeptide lacks a pair of antibody light and heavy chain variable domains specifically binding to a target; and wherein the second CH3 domain comprises a perturbing mutation selected from D356K, E357K, K370E and K439E with corresponding residue requirements in the first CH3 domain; all numbering is according to Kabat EU index.
Multimeric polypeptide with CH3 and variable-domain placement and CH3 perturbing mutation scheme
A multimeric polypeptide comprising a first polypeptide including a first human immunoglobulin G CH3 domain and a first antibody variable domain selected from a pair of an antibody light and heavy chain variable domain specifically binding to a first target; and including a pair of antibody light and heavy chain variable domains specifically binding to a second target either N-terminal to the first CH3 domain or C-terminal to the first variable domain; and a second polypeptide including a second human immunoglobulin G CH3 domain and a second antibody variable domain selected from a pair of an antibody light and heavy chain variable domain specifically binding to a third target; wherein the second CH3 domain comprises a perturbing mutation selected from D356K, E357K, K370E and K439E, whereby the first CH3 domain comprises K at position 439 if D356K, K at position 370 if E357K, E at position 357 if K370E, or D at position 356 if K439E; all numbering is according to Kabat EU index.
Across the independent claims, the main inventive content is the construction of multimeric polypeptides from two polypeptides with defined placement of human IgG CH3 domains and antibody light/heavy variable domains for binding to multiple targets, together with a Kabat EU-numbered CH3 perturbing-mutation scheme in which the second CH3 mutation selection dictates specific residue identities in the first CH3 domain.
Stated Advantages
Supports reduction-free assembly of full-length bi-/multispecific antibodies from destabilized, non-complete antibody formats.
Promotes dissociation and correct assembly of full-length bi-/multispecific antibodies by asymmetric perturbing CH3-domain charge mutations.
Enables physiological/on-surface polypeptide exchange driven by Fc/CH3 interactions without reduction.
Allows chain exchange/assembly using an exchange framework based on CH3 heterodimerization behavior.
Documented Applications
Medicaments and pharmaceutical compositions for treatment use cases are described in connection with the multispecific antibodies and constructs.
On-cell/in vivo assembly context is described for assembling full-length bi-/multispecific antibodies on cells, including constructs involving a cell surface antigen context such as human CD3.
Compositions for pharmaceutical use and use with pharmaceutically acceptable excipients.
Detection/diagnostic uses.
Immunoconjugates including antibody-drug conjugates (ADCs).
Use involving cytotoxic agents.
Pharmaceutical formulations containing the described entities.
Interested in licensing this patent?