Fusion proteins specific for CD137 and GPC3

Patent metadata

Field	Value
Publication number	US-12486330-B2
Application number	US-202017433155-A
Country	US
Kind code	B2
Filing date	Feb 25, 2020
Priority date	Feb 26, 2019
Publication date	Dec 2, 2025
Grant date	Dec 2, 2025

Abstract

The disclosure provides fusion proteins specific for both CD137 and GPC3, which fusion protein can be used to co-stimulate lymphocyte activation in a GPC3-target-dependent manner. Such fusion proteins can be used in many pharmaceutical applications, for example, as anti-cancer agents and/or immune modulators for the treatment or prevention of human diseases such as a variety of tumors. The present disclosure also concerns methods of making the fusion proteins described herein as well as compositions comprising such fusion proteins. The present disclosure further relates to nucleic acid molecules encoding such fusion proteins and to methods for generation of such fusion proteins and nucleic acid molecules. In addition, the application discloses therapeutic and/or diagnostic uses of such fusion proteins as well as compositions comprising one or more of such fusion proteins.

First claim

The invention claimed is:

1 . A fusion protein that is capable of binding both CD137 and GPC3, wherein the fusion protein comprises at least two subunits, wherein a first subunit comprises a full-length immunoglobulin and is specific for GPC3, and wherein a second subunit comprises a lipocalin mutein and is specific for CD137, wherein the second subunit is linked at the N-terminus to the C-terminus of the first subunit via a peptide linker, and wherein the fusion protein comprises the amino acid sequences of SEQ ID NOs: 87 and 82.

2 . The fusion protein of claim 1 ,
(a) wherein the fusion protein is capable of binding GPC3 with a KD value of at most about 1 nM or comparable to or lower than the KD value of the immunoglobulin or an antigen-binding domain thereof that is included in the first subunit alone; (b) wherein the fusion protein is capable of binding CD137 with an EC 50 value of at most about 3 nM or comparable to or lower than the EC 50 value of the lipocalin mutein specific for CD137 that is included in the second subunit alone; (c) wherein the fusion protein is cross-reactive with cynomolgus GPC3; (d) wherein the fusion protein is capable of simultaneously binding CD137 and GPC3 with an EC so value of at most about 10 nM, when said fusion protein is measured in an ELISA assay; (e) wherein the fusion protein is capable of binding GPC3 expressing tumor cells; (f) wherein the fusion protein is capable of inducing increased secretion of IL-2; (g) wherein the fusion protein is capable of inducing increased IL-2 secretion to a higher level than SEQ ID NO: 83 and/or with a better efficiency as compared to SEQ ID NO: 83; (h) wherein the fusion protein is capable of inducing lymphocyte-mediated cytotoxicity; (i) wherein the fusion protein is capable of inducing enhanced killing of GPC3 expressing tumor cells mediated by T cells than SEQ ID NO: 83 and/or inducing cytotoxic T cell-activation with a better efficacy as compared to SEQ ID NO: 83; (j) wherein the fusion protein is capable of co-stimulating T-cell responses in a GPC3-dependent manner; or (k) wherein the fusion protein is capable of co-stimulating T-cell responses in a tumor microenvironment.

3 . The fusion protein of claim 1 , wherein the fusion protein has a half-life in mice of at least 50 hours, at least 75 hours, at least 100 hours, at least 125 hours, at least 150 hours, at least 175 hours, at least 200 hours, at least 250 hours, or even longer, and/or wherein the fusion protein has a half-life in mice that is longer than that of SEQ ID NO: 83.

4 . The fusion protein of claim 1 , wherein the fusion protein has an isoelectric point of at least 6.5, at least 6.8, at least 7.1, at least 7.4, at least 7.5, at least 7.7, or even higher, and/or wherein the fusion protein has an isoelectric point higher than that of SEQ ID NO: 83.

5 . A nucleic acid molecule comprising a nucleotide sequence encoding the fusion protein of claim 1 .

6 . A pharmaceutical composition comprising one or more fusion proteins of claim 1 .

Assignees

• Pieris Pharmaceuticals Gmbh

Inventors

• Bel Aiba Rachida Siham
• Bossenmaier Birgit
• Jaquin Thomas
• Peper-Gabriel Janet
• Hansbauer Eva-Maria
• Schlosser Corinna
• Olwill Shane

Classifications

• C07K2319/00

  Fusion polypeptide · CPC title

• C07K2317/94

  Stability, e.g. half-life, pH, temperature or enzyme-resistance · CPC title

• C07K2317/92

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

• C07K2317/33

  Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity · CPC title

• C07K14/47 Primary

  from mammals · CPC title