Manufacturing optimization of GL-2045, a multimerizing stradomer
Inventors
Block, David S. • MÉRIGEON, Emmanuel Y. • Olsen, Henrik
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11795193-B2
Publication Date
2023-10-24
Expiration Date
Abstract
The present disclosure involves optimized methods for production of biologically active proteins termed optimally manufactured stradomers. The present disclosure further provides compositions and methods useful in the treatment of diseases and conditions including autoimmune diseases, inflammatory diseases, or infectious diseases.
Core Innovation
The invention provides a recombinantly produced homodimer and a multimer of the homodimer, where the homodimer comprises two monomers each comprising amino acids 21-264 of SEQ ID NO:4 and does not comprise an antigen binding domain. The multimer profile is controlled by defined percentage ranges, with the homodimer comprising less than about 20% of the total composition and higher-order homodimer multimers present in specified ranges.
The document describes an optimally manufactured multimerizing stradomer (GL-2045) in which the homodimer and multimer of the homodimer are present as a defined multimer/banding pattern. It further describes producing the homodimer and/or multimer from CHO cell culture media and manufacturing the drug substance through upstream and downstream process controls that preserve the defined multimer profile.
Downstream processing includes protein A purification with pH elution gradient/step elution, followed by ion-exchange and hydrophobic interaction chromatography polishing, and SEC-HPLC assessment of multimer percentages. These processing steps separate very high molecular weight unordered aggregates from lower and higher order multimers and adjust the multimer composition.
Claims Coverage
Independent claim coverage is provided by three independent composition claims that define a recombinantly produced homodimer lacking an antigen binding domain together with a multimer of the homodimer having specific oligomer composition percentage ranges. Across the independent claims, the inventive features include the structural definition of the homodimer monomers, the exclusion of an antigen binding domain, quantitative limits for the homodimer content relative to total composition, and specific percentage ranges for multiple multimer/oligomer size categories.
Defined homodimer monomer region without antigen binding domain
The composition comprises a recombinantly produced homodimer and a multimer of the homodimer, wherein the homodimer comprises two monomers each comprising amino acids 21-264 of SEQ ID NO:4, and the homodimer does not comprise an antigen binding domain.
Homodimer content limited to less than about 20% of total composition
The homodimer comprises less than about 20% of the total composition.
Heptamer-plus abundance and defined multimer oligomer ranges
The multimer of the homodimer comprises a heptamer of the homodimer and above comprising at least about 28% of the total composition, together with specified percentage ranges for dimer through hexamer either individually or as grouped ranges.
Across the independent claims, the composition is defined by a recombinantly produced homodimer consisting of monomers comprising amino acids 21-264 of SEQ ID NO:4 and lacking an antigen binding domain, with the homodimer constituting less than about 20% of the total composition. The multimer is further defined by minimum heptamer-plus content of at least about 28% and by specified percentage ranges for multimer/oligomer states.
Stated Advantages
Controls multimer profile by separating very high molecular weight unordered aggregates from biologically active lower and higher order multimers.
Reduces or eliminates residual high-MW material and specified multimer species by using defined processing conditions.
Achieves defined multimer pattern through an integrated manufacturing protocol using chromatography conditions and multimer analysis after each step.
The manufacturing approach aims to increase the higher-order multimer percentage while maintaining high CHO cell viability and high protein titer.
The defined multimer profile is associated with FcγRIIIA avidity, complement-dependent cytotoxicity inhibition, and chemotaxis inhibition.
Documented Applications
Drug substance manufacturing using an integrated upstream culture context and downstream chromatographic processing to obtain a defined multimer composition for analysis and characterization.
Multimer analysis after each processing step using analytical SEC-HPLC/HPLC to support the defined multimer composition ranges.
Purification and polishing of GL-2045 multimers using protein A affinity chromatography, ion-exchange polishing, and hydrophobic interaction chromatography to control the multimerization profile and reduce unordered high-molecular-weight aggregates.
Treating autoimmune/inflammatory/complement-mediated diseases and infectious diseases.
Interested in licensing this patent?