Multi-specific polypeptide useful for localized tumor immunomodulation

Patent metadata

Field	Value
Publication number	US-10927154-B2
Application number	US-201515111153-A
Country	US
Kind code	B2
Filing date	Jan 12, 2015
Priority date	Jan 13, 2014
Publication date	Feb 23, 2021
Grant date	Feb 23, 2021

Abstract

The disclosure provides a multi-specific polypeptide with a first moiety specific for a tumor-associated antigen on tumor cell surface and a second moiety specific for an immune checkpoint protein, which multi-specific polypeptide can be useful for biasing a T-cell-mediated response to a tumor micro-environment. For example, the polypeptide may contain: a) a first binding domain, for example, a full-length antibody or an antigen-binding domain of an antibody, specifically recognizing a tumor-associated antigen on tumor cell surface, and b) a second binding domain, such as a lipocalin mutein, capable of stimulating T-cell proliferation e.g., by inhibiting a protein receptor that down-regulates the immune system. The first binding domain may be genetically linked (i.e., peptide bond at its N- or C-terminus) to the second binding domain. The multispecific polypeptide also may contain a third or yet additional specific binding moieties, any of which can specifically bind a distinct immune checkpoint protein. The polypeptide may contain an Fc region of an antibody or of an antigen-binding domain thereof and simultaneously engage (1) a T cell receptor complex of a T cell, (2) a tumor-associated antigen on tumor cell surface, while (3) preserving the Fc function of the Fc region to Fc receptor-positive cell. The polypeptide is useful for the induction of an anti-tumor immunity in humans and/or animals. The disclosure also provides thermal-stable lipocalin muteins specific for CTLA-4. The disclosure further relates to a process for the production of the polypeptide or muteins as well nucleic acids encoding for the polypeptide or muteins, to vectors comprising the same and to host cells comprising the vector. In another aspect, the disclosure provides for a pharmaceutical composition comprising the polypeptide or muteins and medical uses of the polypeptide or muteins.

First claim

The invention claimed is:

1. A fusion protein comprising at least two binding domains, wherein a first binding domain comprises a full-length immunoglobulin, wherein a second binding domain comprises a mutein of human neutrophil gelatinase-associated lipocalin (hNGAL)
wherein said first binding domain has binding specificity for Her2/neu, and said second binding domain has binding specificity for CTLA-4;
wherein the fusion protein comprises said second binding domain fused to C-terminus of each heavy chain and each light chain of the first binding domain; and
wherein the fusion protein comprises the amino acid sequences of SEQ ID NOs: 97 and 99.

2. The fusion protein of claim 1 , wherein the fusion protein activates CTLA-4.

3. The fusion protein of claim 1 , wherein the Fc function of the Fc region of the full-length immunoglobulin to Fc receptor-positive cell is preserved.

4. A nucleic acid molecule comprising a nucleotide sequence encoding the fusion protein of claim 1 , wherein the nucleic acid molecule is comprised in a vector or in a phagemid vector, or a host cell containing said nucleic acid molecule.

5. A pharmaceutical composition comprising the fusion protein of claim 1 and a pharmaceutically acceptable excipient.

6. A diagnostic or analytical kit comprising the fusion protein of claim 1 .

Assignees

• Pieris Pharmaceuticals Gmbh

Inventors

• Olwill Shane
• Wiedenmann Alexander
• Allersdorfer Andrea
• Aiba Rachida Bel
• Matschiner Gabriele
• Lunde Bradley

Classifications

• C07K16/2863

  against receptors for growth factors, growth regulators · CPC title

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title

• C07K2318/20

  Antigen-binding scaffold molecules wherein the scaffold is not an immunoglobulin variable region or antibody mimetics · CPC title

• C07K16/32

  against translation products of oncogenes · CPC title

• C07K2317/76

  Antagonist effect on antigen, e.g. neutralization or inhibition of binding · CPC title