Cytokine conjugates
Patent Information
- Authority / Receiving Office
- RU · RU
- Patent Type
- Patents
- Current Assignee / Owner
- AMUNIX PHARMACEUTICALS INC
- Filing Date
- 2021-06-24
- Publication Date
- 2026-06-30
Smart Images
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Abstract
Claims
1. A fusion protein useful for controlled release of a cytokine, comprising a cytokine that is IL-12 or a variant of IL-12 that contains a p35 subunit and a p40 subunit, where each subunit has an N-terminus and a C-terminus, and an extended recombinant polypeptide (XTEN) fused to each of the two N-termini and two C-termini of the cytokine via a release segment (RS), wherein each XTEN has a length of 100 to 3000 amino acid residues and comprises several units of amino acid sequences selected from the group of SEQ ID NOs: 182-203 and 881-888.
2. The fusion protein of claim 1, wherein each release segment (RS) is characterized by at least 88%, at least 94%, or 100% sequence identity with a sequence selected from SEQ ID NOs: 20-151 and 335-830.
3. The fusion protein of claim 1, wherein the p40 subunit comprises an amino acid sequence characterized by at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the sequence under SEQ ID NO:
5.
4. The fusion protein of claim 1, wherein the p35 subunit comprises an amino acid sequence characterized by at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the sequence of SEQ ID NO:
6.
5. The fusion protein of any one of claims 1-4, wherein the cytokine comprises a linker located between the p35 subunit and the p40 subunit.
6. The fusion protein of claim 5, wherein the cytokine is an IL-12 variant comprising an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:
7.
7. The fusion protein of any one of claims 1-6, wherein XTEN comprises one or more motifs selected from the group consisting of SEQ ID NO: 204-221 and 889-908.
8. The fusion protein of any one of claims 1 to 7, wherein the binding activity of the cytokine, when bound to XTEN in the fusion protein, to the corresponding cytokine receptor is characterized by a half-maximal effective concentration (EC50) that is at least 1.2 times greater, at least 1.4 times greater, at least 1.6 times greater, at least 1.8 times greater, at least 2.0 times greater, at least 3.0 times greater, at least 4.0 times greater, at least 5.0 times greater, at least 6.0 times greater, at least 7.0 times greater, at least 8.0 times greater, at least 9.0 times greater, or at least 10.0 times greater than the EC50 characterizing the corresponding binding activity of the cytokine when not bound to XTEN, as determined in an in vitro binding assay.
9. The fusion protein of claim 8, wherein the in vitro binding assay utilizes a genetically engineered cell line with a reporter gene configured to respond to the binding of a cytokine to a corresponding cytokine receptor with proportional expression of the reporter protein.
10. A pharmaceutical composition for use in stimulating an antitumor immune response, comprising an effective amount of a fusion protein according to any one of claims 1-9 and at least one pharmaceutically acceptable carrier.
11. Use of the composition according to claim 10 in the production of a medicinal product for stimulating an antitumor immune response in a subject in need thereof.
12. A method for stimulating an anti-tumor immune response in a subject, the method comprising administering to the subject a therapeutically effective amount of the fusion protein of any one of claims 1-9 or the composition of claim 10.
13. The method of claim 12, further comprising administering to the subject a therapeutically effective amount of at least one immune checkpoint inhibitor.
14. The method according to claim 12 or 13, wherein the fusion protein is administered intravenously, subcutaneously or orally.