Single chain variable fragment CD3 binding proteins
Inventors
Dubridge, Robert B. • SETO, Pui • Austin, Richard J. • Evnin, Luke • Guenot, Jeanmarie • Lemon, Bryan
Assignees
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Publication Number
US-11623958-B2
Publication Date
2023-04-11
Expiration Date
Abstract
Disclosed herein are single chain variable fragment CD3 binding proteins with improved binding affinities, and robust aggregation profiles. Also described are multispecific binding proteins comprising a single chain variable fragment CD3 binding protein according to the instant disclosure. Pharmaceutical compositions comprising the binding proteins disclosed herein and methods of using such formulations are provided.
Core Innovation
The invention relates to scFv CD3-binding proteins defined by specific variable heavy chain (VH) and variable light chain (VL) complementarity determining regions (HC CDR1/HC CDR2/HC CDR3 and LC CDR1/LC CDR2/LC CDR3) sequence patterns with framework residue constraints. The scFv CD3-binding proteins are described as having an amino acid sequence that is at least eighty percent identical to SEQ ID NO: 22 (wt anti-CD3), and in further restrictions as being at least ninety-five percent identical to a sequence selected from a group of SEQ ID NOs including SEQ ID NO: 8-21, 94, and 95.
The document specifies alternative sets of CDR assignments in which HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 are each set forth as particular SEQ ID NOs. It further describes variants based on wt anti-CD3 SEQ ID NO:22 and provides mutation schemes involving CDR and other residue substitutions that yield improved CD3 binding. Binding is described for human CD3 (hKd) and cynomolgus CD3 (cKd) with comparative affinity versus wt anti-CD3.
The invention further includes multispecific constructs that combine the CD3-binding scFv with a serum albumin half-life extension domain and a target antigen binding domain. The document discusses formulation/administration concepts and provides example data for affinity selection, cytotoxicity of CD20×CD3 bispecifics, and thermal stability measurements of anti-CD3ε scFv variants.
Claims Coverage
The provided excerpt contains three independent claims (clm-00001, clm-00020, clm-00024). Each claim centers on administering a CD3-binding scFv with specified VH and VL CDR compositions and SEQ ID-defined sequence identity constraints, with clm-00024 additionally requiring a serum albumin-binding domain and a cancer antigen binding domain.
Cancer treatment using a CD3-binding scFv with defined VH and VL CDR sequences
A method of treating a cancer by administering a protein comprising a single chain variable fragment (scFv) CD3 binding protein with VH and VL, wherein the scFv has HC CDR1/HC CDR2/HC CDR3 and LC CDR1/LC CDR2/LC CDR3 defined by specific SEQ ID NOs, and wherein the scFv CD3 binding protein is at least eighty percent identical to SEQ ID NO: 22, and further has at least ninety-five percent identity to a sequence selected from SEQ ID NOs including SEQ ID NO: 8-21, 94, and 95, the protein further comprising a cancer antigen binding domain.
Cancer treatment with a CD3-binding scFv excluding a defined SEQ ID set and using alternative CDR sequence combinations
A method of treating a cancer by administering a protein where the scFv CD3 binding protein includes HC CDR1/HC CDR2/HC CDR3 and LC CDR1/LC CDR2/LC CDR3, wherein the HC-CDR1, HC-CDR2, HC-CDR3, LC-CDR1, LC-CDR1, and LC-CDR3 are not simultaneously SEQ ID NO. 23, SEQ ID NO. 24, SEQ ID NO. 25, SEQ ID NO. 26, SEQ ID NO. 27, and SEQ ID NO. 28, wherein the scFv CD3 binding protein is at least eighty percent identical to SEQ ID NO. 22, and wherein the protein further comprises a cancer antigen binding domain, with the CD3 binding CDRs selected from enumerated combinations of SEQ ID NO-defined CDRs.
Cancer treatment with a multispecific protein including CD3 scFv, serum albumin binding domain, and cancer antigen binding domain
A method of treating a cancer by administering a multispecific protein comprising a single chain variable fragment CD3 binding protein, a domain that specifically binds to a serum albumin, and a cancer antigen binding domain, wherein the CD3 binding protein has VH with HC CDR1/HC CDR2/HC CDR3 and VL with LC CDR1/LC CDR2/LC CDR3 defined as one of multiple enumerated SEQ ID NO-defined CDR sets.
Across the independent claims, the inventive coverage is directed to treating cancer via administration of a CD3-binding scFv whose VH/VL CDRs are defined by enumerated SEQ ID NO sets and constrained by SEQ ID identity thresholds to SEQ ID NO: 22, with one claim additionally excluding a specific simultaneous SEQ ID configuration and another claim requiring a multispecific format that includes a serum albumin-binding domain and a cancer antigen binding domain.
Stated Advantages
Improved CD3 binding, including comparative affinity versus wt anti-CD3.
Thermal stability measurement (temperature of hydrophobic exposure, T h /DSF) of anti-CD3ε scFv variants is provided in the document.
Documented Applications
Treating a cancer by administering the specified CD3-binding scFv protein or multispecific protein.
Affinity selection and example data related to cytotoxicity of CD20×CD3 bispecifics.
Thermal stability measurements of anti-CD3ε scFv variants (T h /DSF).
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