Search

JP-2026514293-A - A fusion protein of β2-microglobulin, HLA heavy chain polypeptide, and CD47-SIRPα inhibitor.

JP2026514293AJP 2026514293 AJP2026514293 AJP 2026514293AJP-2026514293-A

Abstract

The present invention relates to a fusion protein, the fusion protein comprising: a β2m (beta-2 microglobulin) polypeptide, wherein the β2m polypeptide is linked to an HLA heavy chain polypeptide containing an HLAα3 domain by a β2m linker peptide; optionally, an Fc polypeptide linked to an HLA heavy chain polypeptide via a peptide linker, optionally linked to a polypeptide inhibitor of the interaction between cellular CD47 and cellular signaling regulatory protein alpha (SIRPα) by a short peptide linker. Fusion proteins and their dimers are particularly useful as cancer treatments. [Selection Diagram] None

Inventors

  • ラフィエイ，アナヒタ
  • コーツ，スティーブン
  • エバースバッハ，ヒルマー
  • マロカン ベラウンザラン，オシリス

Assignees

  • イムノス セラピューティクス アーゲー

Dates

Publication Date
20260508
Application Date
20240208
Priority Date
20230208

Claims (20)

  1. It is a fusion protein: a. β2m (beta-2 microglobulin) polypeptide; β2m linker peptide; b. HLA heavy chain polypeptides containing the HLAα3 domain; The Fc polypeptide is optionally connected to the HLA heavy chain polypeptide via a peptide linker, particularly a peptide linker having a length of 5 to 25 aa, and optionally connected to the following polypeptide inhibitors by a short peptide linker: c. The fusion protein comprising a polypeptide inhibitor of the interaction between CD47 and signal regulatory protein alpha (SIRPα).
  2. The β2m polypeptide consists of a single copy of the β2m sequence, and the HLA heavy chain polypeptide consists of an HLA heavy chain composed of an α1 domain, an α2 domain, and an α3 domain. In particular, the fusion protein according to claim 1, wherein the HLA heavy chain polypeptide consists of an HLA-B * 57 heavy chain composed of an α1 domain, an α2 domain, and an α3 domain.
  3. The fusion protein according to claim 2, wherein the β2m polypeptide and the HLA heavy chain polypeptide together are characterized by the sequence of SEQ ID NO: 017, or a sequence having at least (≧) 85% identity with SEQ ID NO: 017 and having at least 75% of the biological function of SEQ ID NO: 017.
  4. The β2m polypeptide consists of a single copy of the β2m sequence, and the HLA heavy chain polypeptide consists of one, two, or three HLAα3 domains. The fusion protein according to claim 1, in particular, the HLA heavy chain polypeptide comprises one, two, or three HLA-B * 57α3 domains.
  5. The fusion protein according to claim 4, wherein the β2m polypeptide and the HLA heavy chain polypeptide together are characterized by the sequence of SEQ ID NO: 019, or a sequence having at least (≧) 85% identity, particularly 90% or more, 93% or more, 96% or more, or 98% or more identity with SEQ ID NO: 019, and having at least 75% of the biological function of SEQ ID NO: 019.
  6. The β2m polypeptide consists of two copies of the β2m sequence linked by an internal (β2m) 2- peptide linker, and the HLA heavy chain polypeptide consists of two HLAα3 domains. In particular, the fusion protein according to claim 1, wherein the HLA heavy chain polypeptide consists of two HLA-B * 57α3 domains.
  7. The aforementioned internal (β2m) 2- peptide linker is a glycine-serine linker composed of 1 to 30 amino acids. The fusion protein according to claim 6, in particular, the internal (β2m) 2 -peptide linker is represented by formula (GGGGS)n (Sequence ID: 003), where n is 1, 2, 3, 4, or 5.
  8. The fusion protein according to claim 6 or 7, wherein the β2m polypeptide and the HLA heavy chain polypeptide together are characterized by the sequence of SEQ ID NO: 021, or a sequence having at least (≧) 85% identity, particularly 90% or more, 93% or more, 96% or more, or 98% or more identity with SEQ ID NO: 021, and having at least 75% of the biological function of SEQ ID NO: 021.
  9. The fusion protein according to any one of claims 4 to 8, wherein the first α3 domain and the second α3 domain are connected by an (α3) 2 peptide linker, the (α3) 2 peptide linker is a glycine-serine linker composed of 1 to 30 amino acids and is represented by an n-mer of (GGGGS) (Sequence ID: 005), where n is 1, 2, 3, 4, or 5.
  10. A β2m linker peptide having a length of 1 to 30 amino acids, particularly 10 to 20 amino acids, is used to link a β2m polypeptide to an HLA heavy chain polypeptide, as described in any one of claims 1 to 9.
  11. The fusion protein according to any one of claims 2 to 11, wherein the HLA heavy chain polypeptide is selected from the group consisting of B * 57 polypeptide, B * 58 polypeptide, B * 27 polypeptide, B * 44 polypeptide, B * 81 polypeptide, C * 08 polypeptide, and C * 12 polypeptide.
  12. The fusion protein according to any one of claims 1, 2, 4 , 6 , 7, 9, or 10, wherein the HLA heavy chain polypeptide is selected from the group consisting of HLA-A * 11 polypeptide, A * 24 polypeptide, A * 25 polypeptide, A * 45 polypeptide, B*78 polypeptide, B * 81 polypeptide, C*06 polypeptide, and C * 12 polypeptide.
  13. The fusion protein according to any one of claims 1 to 11, wherein the HLA heavy chain polypeptide is a B * 57 polypeptide.
  14. The HLA heavy chain polypeptide has the sequence B * 57 of SEQ ID NO: 006, - A fusion protein according to any one of claims 8 to 12, comprising one or more mutations selected from the 7th, 46th, 97th, 143rd, 194th, and 197th positions.
  15. The fusion protein according to any one of claims 8 to 12, wherein the HLA heavy chain polypeptide comprises the A46E and V97R mutations in the B * 57 sequence of SEQ ID NO: 006.
  16. The fusion protein according to any one of claims 8 to 12, wherein the HLA heavy chain polypeptide comprises the A46E, V97R, and I194T mutations in the B * 57 sequence of SEQ ID NO: 006.
  17. The fusion protein according to any one of claims 8 to 12, wherein the HLA heavy chain polypeptide comprises the A46E, V97R, and H197F mutations in the B * 57 sequence of SEQ ID NO: 006.
  18. The fusion protein according to any one of claims 8 to 12, wherein the HLA heavy chain polypeptide comprises the mutations A46E, V97R, I194T and H197F in the sequence of B * 57 of SEQ ID NO: 006.
  19. The fusion protein according to any one of claims 1 to 4, 7, or 10 to 18, wherein the HLA heavy chain polypeptide comprises or consists of a polypeptide selected from SEQ ID NOs: 007, 061, 62, 63, and 64.
  20. The polypeptide inhibitor of the interaction between CD47 and SIRPα is a SIRPα extracellular domain polypeptide, according to any one of claims 1 to 19, for the fusion protein.

Description

This invention relates to a fusion protein and its use in patients diagnosed with cancer. The fusion protein described in this invention comprises β2m, a soluble HLA (human leukocyte antigen) heavy chain polypeptide, and an inhibitor of the interaction between CD47 and signal-regulating protein alpha (SIRPα). This application claims priority to European Patent Application (EP) No. 23155693.7, filed on 8 February 2023, and European Patent Application (EP) No. 23213177.1, filed on 29 November 2023, which are incorporated herein by reference. The tumor microenvironment contains a large number of macrophages, sometimes accounting for up to 50% of the tumor mass. Importantly, macrophages perform immune surveillance. Therefore, both innate immunity and tumor-associated macrophages are gradually gaining attention as targets for innate immunotherapy. However, there is a complex relationship between tumors and macrophages, and in some cases, macrophages can actually promote tumor growth or metastasis. The dual role of macrophages has been shown to stem from their high degree of heterogeneity and plasticity. Macrophages can be broadly classified into two distinct groups: type 1 or classically activated (M1) and type 2 or alternatively activated (M2). In vitro, M1 cells are characterized by a pro-inflammatory phenotype, exhibiting antimicrobial activity and leading to tumor suppression, while M2 macrophages can promote tissue repair, matrix remodeling, and angiogenesis-supporting tumorigenesis. Different macrophage phenotypes result in different phagocytic activities. In the context of cancer, macrophages can rapidly detect membrane molecules on tumor cells and engulf tumor cells through phagocytosis, a multi-step cellular process involving target cell recognition, cell entrapment, and lysosomal digestion, regulated by receptor-ligand interactions between target cells and phagocytic cells. Multiple antiphagocytic signals present in cancer cells have been identified, including LILRB1 and LILRB2. The leukocyte Ig-like receptor subfamily B (LILRB) is a group of type I transmembrane glycoproteins possessing an extracellular Ig-like domain that binds to a ligand, and an intracellular ITIM capable of recruiting tyrosine phosphatases SHP-1, SHP-2, and inositol phosphatase SHIP. Macrophages express both LILRB1 and LILRB2 receptors, and activation of these receptors downregulates macrophage activity in the tumor microenvironment. LILRB1 blockade in immune cells has been demonstrated to be effective against solid and liquid tumors using in vitro models. For example, LILRB1 signaling can inhibit monocyte activation and macrophage phagocytosis (Colonna M. et al. 1997 J. Exp. Med. 186(1):1809). LILRB2 blockade reprograms TAMs into a pro-inflammatory phenotype, suppresses T regulatory cell infiltration, and enhances the effectiveness of immune checkpoint inhibitors. Furthermore, LILRB2 blockade inhibits SHP-1/SHP-2 receptor-mediated activation and enhances pro-inflammatory responses (Alsina-Beauchamp D. et al. 2018, J. Clin. Invest. 128(12):5647). ImmunOs Therapeutics is developing molecules based on human leukocyte antigen (HLA) heavy chains that bind to the LILRB1, LILRB2, and KIR3DL1 receptors (see, for example, International Publication (WO) 2017153438 (A1)). International Publication (WO) No. 2017153438 (A1) Colonna M. et al. 1997 J. Exp. Med. 186(1):1809Alsina-Beauchamp D. et al. 2018, J. Clin. Invest. 128(12):5647 Detailed Description of the Invention A first aspect of the present invention relates to a fusion protein comprising: a. A β2m (beta-2 microglobulin) polypeptide comprising, or consisting of, sequence number 001, or a sequence having at least 85% identity with sequence number 001; A β2m linker peptide that connects a β2m polypeptide and an HLA heavy chain polypeptide, wherein the β2m polypeptide is linked to the HLA heavy chain polypeptide via the β2m linker peptide; b. HLA heavy chain polypeptides containing the HLAα3 domain; The Fc polypeptide is then optionally connected to an HLA heavy chain polypeptide via a peptide linker, particularly a peptide linker having a length of 5 to 25 aa, and this optional Fc polypeptide is linked by a short optional peptide linker to a polypeptide inhibitor of the interaction between cellular CD47 and cellular signal regulatory protein alpha (SIRPα); in other words, this optional Fc polypeptide can act as a bridge between the β2m-HLA domain and the SIRPα-CD47 inhibitory domain, and c. a polypeptide inhibitor of the interaction between cellular CD47 and cellular signal regulatory protein alpha (SIRPα). In a particular embodiment, a single fusion protein would contain all of the above functionalities as part of the same polypeptide sequence. In that particular embodiment, all of the above functionalities, including the Fc polypeptide, would be arranged in the order described, i.e., from the N-terminus to the C-terminus of the fusion protein, in the order of β2m, HLA, Fc, and inhibit