IP Library Granted Patent US 12,240,907
Granted Patent B2
US 12,240,907 · App. 17/088,918 · Granted Mar 4, 2025

N-terminal scFv multispecific binding molecules

Inventors: Yang Shen (Scarsdale, NY); Ann-Hwee Lee (Ardsley, NY); Yan Yang (Mamaroneck, NY); Chia-Yang Lin (Scarsdale, NY)
Assignee: Regeneron Pharmaceuticals, Inc.
C07K16/2863C07K16/2896C07K16/40C07K2317/31C07K2317/35C07K2317/52C07K2317/522C07K2317/54C07K2317/55C07K2317/565C07K2317/622C07K2317/92
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Quick Facts
Patent No.
US 12,240,907
App. No.
17/088,918
Granted
Mar 4, 2025
Kind
B2
Abstract

Multispecific binding molecules (MBMs) comprising an N-terminal scFv, a first Fab and a second Fab, MBM conjugates comprising the MBMs and cytotoxic or cytostatic agents, pharmaceutical compositions containing the MBMs and MBM conjugates, methods of using the MBMs, MBM conjugates and pharmaceutical compositions for treating cancer, nucleic acids encoding the MBMs, cells engineered to express the MBMs, and methods of producing MBMs.

Claims (22)

1. A method, comprising contacting a cell expressing fibroblast growth factor receptor 3 (FGFR3) and amyloid precursor-like protein 2 (APLP2) with a multispecific binding molecule (MBM) comprising:

(a) a first polypeptide chain comprising, in an N- to C-terminal orientation, (i) an scFv comprising means for binding human APLP2, (ii) a first heavy chain region of a first Fab comprising means for binding human FGFR3, and (iii) a first Fc domain;

(b) a second polypeptide chain comprising, in an N- to C-terminal orientation, (i) a second heavy chain region of a second Fab comprising means for binding human FGFR3 and (ii) a second Fc domain;

(c) a third polypeptide chain comprising a first light chain that pairs with the first heavy chain region to form the first Fab; and

(d) a fourth polypeptide chain comprising a second light chain that pairs with the second heavy chain region to form the second Fab.

2. The method of claim 1 , wherein the scFv is linked to the first heavy chain region via a linker.

3. The method of claim 2 , wherein the linker is 5 amino acids to 50 amino acids in length.

4. The method of claim 2 , wherein the linker is 7 amino acids to 50 amino acids in length.

5. The method of claim 2 , wherein the linker is 5 amino acids to 20 amino acids in length.

6. The method of claim 2 , wherein the linker is 7 amino acids to 20 amino acids in length.

7. The method of claim 2 , wherein the linker comprises a multimer of GnS (SEQ ID NO: 61 ) or SGn (SEQ ID NO: 62 ), wherein n is an integer from 1 to 7.

8. The method of claim 2 , wherein the linker comprises a multimer of G 4 S (SEQ ID NO: 4).

9. The method of claim 1 , wherein the first Fc domain and the second Fc domain form a heterodimer.

10. The method of claim 9 , wherein the first Fc domain comprises a knob mutation as compared to a wild type Fc domain and the second Fc domain comprises a hole mutation as compared to a wild type Fc domain.

11. The method of claim 9 , wherein the first Fc domain comprises a hole mutation as compared to a wild type Fc domain and the second Fc domain comprises a knob mutation as compared to a wild type Fc domain.

12. The method of claim 1 , wherein the first light chain and the second light chain are universal light chains.

13. The method of claim 1 , wherein the scFv has the configuration, in an N- to C-terminal orientation, VL-linker-VH.

14. The method of claim 1 , wherein the scFv has the configuration, in an N- to C-terminal orientation, VH-linker-VL.

15. The method of claim 1 , wherein the first polypeptide chain further comprises, between the heavy chain region of the first Fab and the first Fc domain, a first hinge domain.

16. The method of claim 14 , wherein the second polypeptide chain further comprises, between the heavy chain region of the second Fab and the second Fc domain, a second hinge domain.

17. The method of claim 1 , wherein the cell is in vitro.

18. The method of claim 1 , wherein the cell is a cancer cell.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jul 7, 2021
From: SHEN, YANG; LEE, ANN-HWEE; YANG, YAN; LIN, CHIA-YANG
To: REGENERON PHARMACEUTICALS, INC.
Reel/Frame 056778/0060 →
Continuity (2)
Provisional Application 62930916 · Nov 5, 2019
Related Publication 20210130477A1 · May 6, 2021
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