Multi-specific binding proteins for cancer treatment

Patent metadata

Field	Value
Publication number	US-12528879-B2
Application number	US-202318327456-A
Country	US
Kind code	B2
Filing date	Jun 1, 2023
Priority date	Jun 9, 2018
Publication date	Jan 20, 2026
Grant date	Jan 20, 2026

Abstract

The present invention relates to novel DLL3/CD3 binding proteins. The invention also relates to nucleic acids encoding such proteins; to methods for preparing such proteins; to host cells expressing or capable of expressing such proteins; to compositions comprising such proteins; and to uses of such proteins or such compositions, in particular for therapeutic purposes in the field of cancer diseases.

First claim

The invention claimed is:

1 . A protein comprising a first antigen binding unit specifically binding to the membrane proximal domain of DLL3 and a second antigen binding unit specifically binding to CD3, wherein said first antigen binding unit specifically binding to DLL3 is selected from the group consisting of i) to iii):
i) an antigen binding unit comprising light chain CDRs comprising the amino acid sequences of SEQ ID NO:1 (CDR1), SEQ ID NO:2 (CDR2) and SEQ ID NO:3 (CDR3) and heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 4 (CDR1), SEQ ID NO:5 (CDR2) and SEQ ID NO:6 (CDR3); ii) an antigen binding unit comprising light chain CDRs comprising the amino acid sequences of SEQ ID NO:7 (CDR1), SEQ ID NO:8 (CDR2) and SEQ ID NO:9 (CDR3) and heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 10 (CDR1), SEQ ID NO:11 (CDR2) and SEQ ID NO:12 (CDR3); and iii) an antigen binding unit comprising light chain CDRs comprising the amino acid sequences of SEQ ID NO:13 (CDR1), SEQ ID NO:14 (CDR2) and SEQ ID NO:15 (CDR3) and heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 16 (CDR1), SEQ ID NO:17 (CDR2) and SEQ ID NO:18 (CDR3).

2 . The protein of claim 1 , wherein said first antigen binding unit specifically binding to the membrane proximal domain of DLL3 comprises a first light chain variable domain and a first heavy chain variable domain and is selected from the group consisting of i) to iii):
i) an antigen binding unit comprising a light chain variable domain comprising the amino acid sequence of SEQ ID NO:37 and heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:38; ii) an antigen binding unit comprising a light chain variable domain comprising the amino acid sequences of SEQ ID NO:39 and heavy chain variable domain comprising the amino acid sequences of SEQ ID NO:40; and iii) an antigen binding unit comprising a light chain variable domain comprising the amino acid sequence of SEQ ID NO:41 and heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:42.

3 . The protein of claim 1 , wherein said second antigen binding unit specifically binding to CD3 is selected from the group consisting of i)-iii):
i) an antigen binding unit comprising light chain CDRs comprising the amino acid sequences of SEQ ID NO:55 (CDR1), SEQ ID NO:56 (CDR2) and SEQ ID NO:57 (CDR3) and heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 58 (CDR1), SEQ ID NO:59 (CDR2) and SEQ ID NO:60 (CDR3); ii) an antigen binding unit comprising light chain CDRs comprising the amino acid sequences of SEQ ID NO:61 (CDR1), SEQ ID NO:62 (CDR2) and SEQ ID NO:63 (CDR3) and heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 64 (CDR1), SEQ ID NO:65 (CDR2) and SEQ ID NO:66 (CDR3); and iii) an antigen binding unit comprising light chain CDRs comprising the amino acid sequences of SEQ ID NO:96 (CDR1), SEQ ID NO:97 (CDR2) and SEQ ID NO:98 (CDR3) and heavy chain CDRs comprising the amino acid sequences of SEQ ID NO: 99 (CDR1), SEQ ID NO:100 (CDR2) and SEQ ID NO:101 (CDR3).

4 . The protein of claim 1 , wherein said second antigen binding unit specifically binding to CD3 comprises a second light chain variable domain and a second heavy chain variable domain selected from the group consisting of i) to iii):
i) an antigen binding unit comprising a light chain variable domain comprising the amino acid sequence of SEQ ID NO:67 and a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:68; ii) an antigen binding unit comprising a light chain variable domain comprising the amino acid sequence of SEQ ID NO:69 and a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:70; and iii) an antigen binding unit comprising a light chain variable domain comprising the amino acid sequence of SEQ ID NO:102 and a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:103.

5 . The protein of claim 1 , wherein
i) said first antigen binding unit specifically binding to the membrane proximal domain of DLL3 comprises from its N to C-terminus a first light chain variable domain, a first light chain constant domain, a first peptide linker, a first heavy chain variable domain and a first heavy chain constant CH1 domain; and ii) said second antigen binding unit specifically binding to CD3 comprises from its N to C-terminus a second light chain variable domain, a second light chain constant domain, a second peptide linker, a second heavy chain variable domain and a second heavy chain constant CH1 domain.

6 . The protein of claim 5 , wherein said first and/or second peptide linker comprises 26 to 42 amino acids, 30 to 40 amino acids, 34 to 40 amino acids, or 36 to 39 amino acids.

7 . The protein of claim 5 , wherein said first peptide linker and/or second peptide linker is a Glycine-Serine linker, comprising the amino acid sequence of SEQ ID NO:89, and wherein said first and second peptide linker comprise the same sequence.

8 . The protein of claim 5 , further comprising a first human IgG1 Fc domain and a second human IgG1 Fc domain, said first human IgG1 Fc domain covalently linked to said first antigen binding unit, and a said second human IgG1 Fc domain covalently linked to said second antigen binding unit.

9 . The protein of claim 8 , wherein
i) said first human IgG1 Fc domain comprises a tyrosine (Y) at position 366 [T366Y], and said second human IgG1 Fc domain comprises a threonine (T) at position 407 [Y407T], or ii) said first human IgG1 Fc domain comprises a tryptophan (W) at position 366 [T366W], and said second human IgG1 Fc domain comprises a serine(S) at position 366 [T366S], an alanine (A) at position 368 [L368A] and a valine (V) at position 407 [Y407V], or iii) said second human IgG1 Fc domain comprises a tyrosine (Y) at position 366 [T366Y], and said first human IgG1 Fc domain comprises a threonine (T) at position 407 [Y407T], or iv) said second human IgG1 Fc domain comprises a tryptophan (W) at position 366 [T366W], and said first human IgG1 Fc domain comprises a serine(S) at position 366 [T366S], an alanine (A) at position 368 [L368A] and a valine (V) at position 407 [Y407V], wherein said first or said second human IgG1 Fc domain optionally further comprises an arginine at position 435 [H435R] and a phenylalanine at position 436 [Y436F], wherein numbering of the amino acids of the human IgG1 Fc domain are according to the EU numbering system.

10 . The protein of claim 5 , wherein the first light chain constant domain and the second light chain constant domain comprise a human kappa or lambda domain.

11 . A method of manufacturing a protein of claim 1 , comprising
i) cultivating a host cell transfected with an expression vector comprising a nucleic acid molecule encoding a first polypeptide chain for the first antigen binding unit specifically binding to the membrane proximal domain of DLL3, and an expression vector comprising a nucleic acid molecule encoding a second polypeptide chain for the second antigen binding unit specifically binding to CD3 under conditions allowing expression of the protein of claim 1 ; and, ii) recovering the protein; and optionally iii) further purifying and/or modifying and/or formulating the protein.

12 . A pharmaceutical composition comprising the protein of claim 1 and a pharmaceutically acceptable carrier.

13 . The protein of claim 1 , wherein
(i) said first antigen binding unit comprises light chain CDRs comprising the amino acid sequences of SEQ ID NO:1 (CDR1), SEQ ID NO:2 (CDR2) and SEQ ID NO:3 (CDR3) and heavy chain CDRs comprising the amino acid sequences of SEQ

Assignees

• Boehringer Ingelheim Int

Inventors

• Hipp Susanne
• Adam Paul
• Dziegelewski Michael
• Ganesan Rajkumar
• Gorman Philip Nicholas
• Gupta Pankaj
• Gupta Priyanka
• Scheer Justin
• Voynov Vladimir H

Classifications

• A61K2039/505

  comprising antibodies · CPC title

• C07K2319/00

  Fusion polypeptide · CPC title

• C07K2317/56

  variable (Fv) region, i.e. VH and/or VL · CPC title

• C12N15/63

  Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression · CPC title

• C07K2317/31

  multispecific · CPC title