Fusion proteins of natural human protein fragments to create orderly multimerized immunoglobulin Fc compositions
Inventors
Block, David S. • Olsen, Henrik
Assignees
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Publication Number
US-11117940-B2
Publication Date
2021-09-14
Expiration Date
Abstract
The current invention involves a series of fully recombinant multimerized forms of immunoglobulin Fc which thereby present polyvalent immunoglobulin Fc to immune cell receptors. The fusion proteins exist as both homodimeric and highly ordered multimeric fractions, termed stradomers. In comparison to the homodimeric fraction, purified multimeric stradomers have higher affinity and avidity for FcγRs with slower dissociation and are useful in the treatment and prevention of disease. The current invention demonstrates that directly linking IgG1 Fc regions to multimerization domains leads to enhanced multimerization and biological activity.
Core Innovation
The invention relates to recombinant, naturally linked stradomer fusion proteins that form orderly homodimeric and highly ordered multimeric Fc multimers. A homodimeric stradomer unit includes two stradomer unit monomers, and the multimerized compound comprises two or more homodimeric stradomer units assembled from these units.
The stradomer design directly fuses IgG Fc to a single multimerization domain to create an ordered multimeric Fc architecture. Purified multimeric stradomers show increased affinity/avidity for FcγRs, including FcγRIIIa and FcγRIIb, together with slower dissociation. The document contrasts this with earlier stradomers that contained extraneous linker/restriction/tag sequences and reports improved biological activity versus those earlier constructs.
The document also describes multimerized compounds comprising two or more homodimeric stradomer units, where each stradomer unit monomer is defined by amino acid residues 21-264 of SEQ ID NO:4, by an SEQ ID NO:4 form lacking amino acid residues 1-20 corresponding to the leader sequence of SEQ ID NO:1, or by direct carboxy terminal linkage between amino acid sequences of SEQ ID NO:2 and SEQ ID NO:3. It also links stradomers to FcγR aggregation or cross-linking and Fc:FcγR signaling, and describes pharmaceutical compositions that include the multimerized compound and higher order multimers.
Claims Coverage
The independent claims collectively cover multimerized compounds comprising two or more homodimeric stradomer units and stradomer unit monomers defined by specific sequence specifications, including residue range specification, leader-sequence-lacking variants, and direct carboxy terminal linkages between defined sequences. Across these independent claims, the core inventive structure centers on homodimeric stradomer units forming multimerized compounds, with selected claim families also supporting pharmaceutical compositions including higher order multimers.
Multimerized compound with homodimeric stradomer units defined by SEQ ID NO:4 (residues 21-264)
A multimerized compound comprising two or more homodimeric stradomer units, wherein each homodimeric stradomer unit comprises two stradomer unit monomers, and wherein each stradomer unit monomer comprises amino acid residues 21-264 of SEQ ID NO:4.
Multimerized compound with homodimeric stradomer units defined by SEQ ID NO:4 lacking leader sequence residues 1-20
A multimerized compound comprising two or more homodimeric stradomer units, wherein each homodimeric stradomer unit comprises two stradomer unit monomers, and wherein each stradomer unit monomer comprises the amino acid sequence of SEQ ID NO:4 lacking amino acid residues 1-20 corresponding to the leader sequence of SEQ ID NO:1.
Multimerized compound with carboxy-terminal direct linkage between SEQ ID NO:2 and SEQ ID NO:3
A multimerized compound comprising two or more homodimeric stradomer units, wherein each homodimeric stradomer unit comprises two stradomer unit monomers, each stradomer unit monomer comprising the amino acid sequence of SEQ ID NO:2 directly linked at its carboxy terminus to the amino acid sequence of SEQ ID NO:3.
Stradomer unit monomer with direct carboxy-terminal linkage of SEQ ID NO:2 to a SEQ ID NO:3 polypeptide
A stradomer unit monomer comprising the amino acid sequence of SEQ ID NO:2 directly linked at its carboxy terminus to a polypeptide comprising the amino acid sequence of SEQ ID NO:3.
Stradomer unit monomer containing amino acid residues 21-264 of SEQ ID NO:4
A stradomer unit monomer comprising amino acid residues 21-264 of SEQ ID NO:4.
Stradomer unit monomer consisting of amino acid residues 21-264 of SEQ ID NO:4
A stradomer unit monomer consisting of amino acid residues 21-264 of SEQ ID NO:4.
Pharmaceutical compositions including higher order multimers
Pharmaceutical compositions that include the multimerized compound and additionally comprise higher order multimers.
Across the independent claims, the claimed subject matter is defined by multimerized compounds comprising two or more homodimeric stradomer units, where each monomer is specified using either amino acid residues 21-264 of SEQ ID NO:4, an SEQ ID NO:4 leader-sequence-lacking form, or direct carboxy-terminal linkage between amino acid sequences of SEQ ID NO:2 and SEQ ID NO:3. Selected independent-claim families also support pharmaceutical compositions that include higher order multimers.
Stated Advantages
Reduces confounding Fc-mediated effects in immunological assays by using stradomers as controls.
Binds endotoxin with very high affinity and can be used as an endotoxin-binding pool.
Provides Fc-FcγR cross-linking that contributes to biological effects.
Increased affinity/avidity for FcγRs, including FcγRIIIa and FcγRIIb.
Slower dissociation from FcγRs.
Improved biological activity versus earlier stradomers containing extraneous linker/restriction/tag sequences.
Improved efficacy in collagen-induced arthritis compared with predecessor constructs.
Synergistic effect with prednisolone at low doses.
Addresses limitations of IVIG.
Documented Applications
Treating or prophylaxis in cancers, autoimmune diseases such as ITP and experimental autoimmune neuritis, and inflammatory or other conditions.
Serving as controls in immunological assays to reduce confounding Fc-mediated effects, including Mixed Lymphocyte Reaction and flow cytometry contexts described in the document.
Using stradomers to bind endotoxin and remove endotoxin complexes.
Application in models including collagen-induced arthritis, ITP prevention, and experimental autoimmune neuritis.
Using stradomers with FcγR aggregation/cross-linking and Fc:FcγR signaling in connection with biological effects described in the document.
Coating/stabilizing Fc reagents on implantable medical devices and other substrates including platelet substrates and FcγR-expressing cells.
Therapeutic and prophylactic uses for immune-mediated inflammatory, autoimmune, and infectious diseases.
Fc-blocking reagents in immunological assays.
Endotoxin binding/removal.
Treatment in collagen-induced arthritis.
Treatment in idiopathic thrombocytopenia purpura (ITP).
Treatment in experimental autoimmune neuritis (EAN).
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