IP Library › Granted Patent US 12,649,796
Granted Patent B2
US 12,649,796 · App. 19/035,322 · Granted Jun 9, 2026

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 12,649,796
App. No.
19/035,322
Granted
Jun 9, 2026
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 (46)

1 . A trivalent 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 to human klotho beta (KLB);

(ii) a linker;

(iii) a first heavy chain region of a first Fab comprising means for binding to human fibroblast growth factor receptor 1c (FGFR1c); and

(iv) a first Fc domain; and

(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 to human KLB; and

(ii) a second Fc domain, wherein the second Fc domain forms a heterodimer with the first 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 trivalent MBM of claim 1 , wherein the linker is at least 5 amino acids in length.

3 . The trivalent MBM of claim 2 , wherein the linker is up to 40 amino acids in length.

4 . The trivalent MBM of claim 3 , wherein the linker is 25 to 35 amino acids in length.

5 . The trivalent MBM of claim 3 , wherein the linker is or comprises a multimer of G n S (SEQ ID NO: 61) or SG n (SEQ ID NO: 62), where n is an integer from 1 to 7.

6 . The trivalent MBM of claim 5 , wherein the linker is or comprises a multimer of G 4 S (SEQ ID NO: 4).

7 . The trivalent MBM of claim 6 , wherein the linker is or comprises SEQ ID NO:63.

8 . The trivalent MBM of claim 6 , wherein the linker is or comprises SEQ ID NO:1.

9 . The trivalent MBM of claim 6 , wherein the linker is or comprises SEQ ID NO:57.

10 . The trivalent MBM of claim 6 , wherein the linker is or comprises SEQ ID NO:64.

11 . The trivalent MBM of claim 6 , wherein the linker is or comprises SEQ ID NO:59.

12 . The trivalent MBM of claim 6 , wherein the linker is or comprises SEQ ID NO:65.

13 . The trivalent MBM of claim 1 , wherein the Fc domains in the Fc heterodimer comprise knob-in-hole mutations as compared to a wild type Fc domain.

14 . The trivalent MBM of claim 1 , wherein at least one Fc domain in the Fc heterodimer comprises a star mutation as compared to a wild type Fc domain.

15 . The trivalent MBM of claim 1 , wherein at least one Fc domain in the Fc heterodimer comprises an amino acid substitution at position P329.

16 . The trivalent MBM of claim 15 , wherein the amino acid substitution is P329A or P329G.

17 . The trivalent MBM of claim 15 , wherein the Fc domain in the Fc heterodimer comprises amino acid substitutions at positions L234 and L235.

18 . The trivalent MBM of claim 16 , wherein the Fc domain in the Fc heterodimer further comprises amino acid substitutions at positions L234 and L235.

19 . The trivalent MBM of claim 16 , wherein the Fc domain in the Fc heterodimer comprises the amino acid mutations L234A, L235A and P329G.

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

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

22 . The trivalent MBM 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.

23 . The trivalent MBM of claim 1 , 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.

24 . The trivalent MBM of claim 1 , wherein the linker is attached to the first heavy chain region of the first Fab.

25 . A composition comprising the trivalent MBM of claim 1 and an excipient.

26 . A method, comprising contacting a cell expressing KLB and FGFR1c with a trivalent MBM comprising:

(a) a first polypeptide chain comprising in an N-to C-terminal orientation:

(i) an scFv comprising means for binding to KLB;

(ii) a linker;

(iii) a first heavy chain region of a first Fab comprising means for binding to human FGFR1c; and

(iv) a first Fc domain; and

(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 to human KLB; and

(ii) a second Fc domain, wherein the second Fc domain forms a heterodimer with the first 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.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 17, 2025
From: SHEN, YANG; LEE, ANN-HWEE; YANG, YAN; LIN, CHIA-YANG
To: REGENERON PHARMACEUTICALS, INC.
Reel/Frame 070868/0412 →
Continuity (4)
Division 17726950 · Apr 22, 2022
Continuation PCTUS2020058798 · Nov 4, 2020
Provisional Application 62930916 · Nov 5, 2019
Related Publication 20250171545A1 · May 29, 2025
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