Multi-chain chimeric polypeptides and uses thereof

Patent metadata

Field	Value
Publication number	US-12479899-B2
Application number	US-202318354866-A
Country	US
Kind code	B2
Filing date	Jul 19, 2023
Priority date	Jun 21, 2019
Publication date	Nov 25, 2025
Grant date	Nov 25, 2025

Abstract

The present disclosure relates to methods that include the use of a first multi-chain chimeric polypeptide and a second multi-chain chimeric polypeptide for stimulating the NK cells, inducing or increasing proliferation of the NK cells, inducing differentiation of the NK cells, and treating a subject in need thereof using activated NK cells.

First claim

What is claimed is:

1 . A method of enhancing cytotoxicity of a NK cell comprising the following steps:
(a) contacting a natural killer cell in a liquid culture medium comprising an effective amount of a first multi-chain chimeric polypeptide for 15 minutes to one day under conditions that allow for differentiation of the natural killer (NK) cell; and (b) contacting the natural killer cell in a liquid culture medium comprising an effective amount of (i) a second multi-chain chimeric polypeptide and (ii) an IgG1 antibody construct, for 1 day to 30 days under conditions that allow for the activation and proliferation of the natural killer cell, wherein the first multi-chain chimeric polypeptide comprises: a first chimeric polypeptide comprising a first target-binding domain, a soluble tissue factor domain comprising a sequence that is at least 90% identical to SEQ ID NO: 5, and a first domain of a pair of affinity domains comprising a sequence that is at least 90% identical to SEQ ID NO: 26; and a second chimeric polypeptide comprising a second domain of a pair of affinity domains comprising a sequence that is at least 90% identical to SEQ ID NO: 24 and a second target-binding domain, wherein the first chimeric polypeptide and the second chimeric polypeptide in the first multi-chain chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains in the first multi-chain chimeric polypeptide; wherein (A) the first target-binding domain in the first multi-chain chimeric polypeptide comprises a sequence at least 90% identical to SEQ ID NO: 20 and the second target-binding domain in the first multi-chain chimeric polypeptide comprises a first sequence that is at least 90% identical to SEQ ID NO: 14 and a second sequence that is at least 90% identical to SEQ ID NO: 16, or (B) the first target-binding domain in the first multi-chain chimeric polypeptide comprises a first sequence that is at least 90% identical to SEQ ID NO: 14 and a second sequence that is at least 90% identical to SEQ ID NO: 16, and the second target-binding domain in the first multi-chain chimeric polypeptide comprises a sequence at least 90% identical to SEQ ID NO: 20; and wherein the second multi-chain chimeric polypeptide comprises: a first chimeric polypeptide comprising a first target-binding domain, a soluble tissue factor domain comprising a sequence that is at least 90% identical to SEQ ID NO: 5, a first domain of a pair of affinity domains comprising a sequence that is at least 90% identical to SEQ ID NO: 26, and a second chimeric polypeptide comprising a second domain of a pair of affinity domains comprising a sequence that is at least 90% identical to SEQ ID NO: 24, and a second target-binding domain, wherein the first chimeric polypeptide and the second chimeric polypeptide in the second multi-chain chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains in the second multi-chain chimeric polypeptide; wherein (A) the first target-binding domain in the second multi-chain chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO: 23 and the second target-binding domain in the second multi-chain chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO: 22, or (B) the first target-binding domain in the second multi-chain chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO: 22 and the second target-binding domain in the second multi-chain chimeric polypeptide comprises a sequence that is at least 90% identical to SEQ ID NO: 23; and wherein the IgG1 antibody construct comprises at least one antigen-binding domain that binds specifically to the soluble tissue factor domain.

2 . The method of claim 1 , wherein the first target-binding domain and the soluble tissue factor domain directly abut each other in the first chimeric polypeptide in the first multi-chain chimeric polypeptide.

3 . The method of claim 1 , wherein the first chimeric polypeptide in the first multi-chain chimeric polypeptide further comprises a linker sequence between the first target-binding domain and the soluble tissue factor domain in the first chimeric polypeptide.

4 . The method of claim 1 , wherein the soluble tissue factor domain and the first domain of the pair of affinity domains in the first chimeric polypeptide in the first multi-chain chimeric polypeptide directly abut each other in the first chimeric polypeptide.

5 . The method of claim 1 , wherein the first chimeric polypeptide in the first multi-chain chimeric polypeptide further comprises a linker sequence between the soluble tissue factor domain and the first domain of the pair of affinity domains in the first chimeric polypeptide.

6 . The method of claim 1 , wherein the second domain of the pair of affinity domains and the second target-binding domain in the second chimeric polypeptide in the first multi-chain chimeric polypeptide directly abut each other in the second chimeric polypeptide.

7 . The method of claim 1 , wherein the second chimeric polypeptide in the first multi-chain chimeric polypeptide further comprises a linker sequence between the second domain of the pair of affinity domains and the second target-binding domain in the second chimeric polypeptide.

8 . The method of claim 1 , wherein the soluble tissue factor domain is a soluble human tissue factor domain.

9 . The method of claim 8 , wherein the soluble human tissue factor domain does not comprise one or more of:
a lysine at an amino acid position that corresponds to amino acid position 20 of mature wildtype human tissue factor protein; an isoleucine at an amino acid position that corresponds to amino acid position 22 of mature wildtype human tissue factor protein; a tryptophan at an amino acid position that corresponds to amino acid position 45 of mature wildtype human tissue factor protein; an aspartic acid at an amino acid position that corresponds to amino acid position 58 of mature wildtype human tissue factor protein; a tyrosine at an amino acid position that corresponds to amino acid position 94 of mature wildtype human tissue factor protein; an arginine at an amino acid position that corresponds to amino acid position 135 of mature wildtype human tissue factor protein; and a phenylalanine at an amino acid position that corresponds to amino acid position 140 of mature wildtype human tissue factor protein.

10 . The method of claim 1 , wherein the first multi-chain chimeric polypeptide and the second multi-chain chimeric polypeptide do not stimulate coagulation in a mammal.

11 . The method of claim 1 , wherein the first target-binding domain and the linker domain in the first chimeric polypeptide in the second multi-chain chimeric polypeptide directly abut each other in the first chimeric polypeptide.

12 . The method of claim 1 , wherein the first chimeric polypeptide in the second multi-chain chimeric polypeptide further comprises a linker sequence between the first target-binding domain and the linker domain in the first chimeric polypeptide.

13 . The method of claim 1 , wherein the linker domain and the first domain of the pair of affinity domains directly abut each other in the first chimeric polypeptide in the second multi-chain chimeric polypeptide.

14 . The method of claim 1 , wherein the first chimeric polypeptide in the second multi-chain chimeric polypeptide further comprises a linker sequence between the linker domain and the first domain of the pair of affinity domains in the first chimeric polyp

Assignees

• Immunitybio Inc

Inventors

• Wong Hing C

Classifications

• A61K40/42

  Cancer antigens · CPC title

• A61K40/15

  Natural-killer [NK] cells; Natural-killer T [NKT] cells · CPC title

• A61K2239/38

  characterised by the dose, timing or administration schedule · CPC title

• C12N5/0646 Primary

  Natural killers cells [NK], NKT cells · CPC title

• A61K2239/31

  characterized by the route of administration · CPC title