Multispecific antibodies and uses thereof

Patent metadata

Field	Value
Publication number	US-2025129144-A1
Application number	US-202318846151-A
Country	US
Kind code	A1
Filing date	Mar 10, 2023
Priority date	Mar 11, 2022
Publication date	Apr 24, 2025
Grant date	—

Abstract

Provided are multispecific antibodies or antigen binding fragments thereof comprising at least one first antigen-binding region capable of binding specifically to pyroglutamate amyloid-β, a second antigen-binding region capable of binding specifically to transferrin receptor (TfR), and a third antigen-binding region capable of binding specifically to paired helical filament (PHF)-tau. Also provided are methods of treating or detecting a neurological disorder and/or delivering a therapeutic or diagnostic agent across the blood-brain barrier. Also described are nucleic acids encoding the antibodies, vectors comprising the nucleic acids, recombinant host cells comprising the nucleic acids and/or vectors, and methods of producing the multispecific antibodies or antigen binding fragments thereof.

First claim

What is claimed is:

1 . A multispecific antibody or antigen binding fragment thereof comprising a first antigen-binding region capable of binding specifically to pyroglutamate amyloid-β, a second antigen-binding region capable of binding specifically to transferrin receptor (TfR), and a third antigen-binding region capable of binding specifically to paired helical filament (PHF)-tau, wherein:
a. the first antigen-binding region comprises:
i. a first heavy chain variable region (VH1) comprising heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 8 or 16, 9 or 17, and 10, respectively; and
ii. a first light chain variable region (VL1) comprising light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 11, 12, and 13, respectively;
b. the second antigen-binding region comprises:
i. a second heavy chain variable region (VH2) comprising heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 1, 2, and 3, respectively; and
ii. a second light chain variable region (VL2) comprising light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 4, 5, and 6, respectively; and
c. the third antigen-binding region comprises:
i. a third heavy chain variable region (VH3) comprising heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 28, 29, and 30, respectively; and
ii. a third light chain variable region (VL3) comprising light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 31, 32, and 33, respectively.

2 . The multispecific antibody or antigen binding fragment thereof of claim 1 , wherein the VH1 comprises an amino acid sequence at least 90% identical to SEQ ID NO:14; and the VL1 comprises an amino acid sequence at least 90% identical to SEQ ID NO:15.

3 . The multispecific antibody or antigen binding fragment thereof of claim 1 , wherein the VH1 comprises the amino acid sequence of SEQ ID NO:14; and the VL1 comprises the amino acid sequence of SEQ ID NO:15.

4 . The multispecific antibody or antigen binding fragment thereof of claim 1 , wherein the second antigen-binding region comprises a single chain fragment variable (scFv) antibody or antigen binding fragment thereof comprising the VH2 and VL2.

5 . The multispecific antibody or antigen binding fragment thereof of claim 4 , wherein the scFv comprises an amino acid sequence at least 90% identical to SEQ ID NO:7.

6 . The multispecific antibody or antigen binding fragment thereof of claim 5 , wherein the scFV comprises the amino acid sequence of SEQ ID NO:7.

7 . The multispecific antibody or antigen binding fragment thereof of claim 1 , comprising:
(j) a first heavy chain (HC1) comprising the VH3 and the VL3, a first heavy chain constant region comprising a first Fc region (Fc1), and the scFv (iv) a second heavy chain (HC2) comprising the VH1 and a second heavy chain constant region comprising a second Fc region (Fc2); (v) a first light chain (LC) comprising the VL1 and a light chain constant region.

8 . The multispecific antibody or antigen binding fragment thereof of claim 7 , wherein the scFv is linked to the carboxy terminus of the first heavy chain constant region via a linker, more particularly a linker comprising the amino acid sequence of SEQ ID NO:27.

9 . The multispecific antibody or antigen binding fragment thereof of claim 7 , wherein the Fc1 and Fc2 each comprise one or more heterodimeric mutations, such as a first and a second modified heterodimeric CH3 domains, respectively, as compared to a wild-type Fc region; particularly, the Fc1 comprises amino acid modifications at positions T350, L351, F405, and Y407, and the Fc2 comprises amino acid modifications at positions T350, T366, K392 and T394, wherein the amino acid modification at position T350 is T350V, T350I, T350L or T350M; the amino acid modification at position L351 is L351Y; the amino acid modification at position F405 is F405A, F405V, F405T or F405S; the amino acid modification at position Y407 is Y407V, Y407A or Y407I; the amino acid modification at position T366 is T366L, T366I, T366V or T366M, the amino acid modification at position K392 is K392F, K392L or K392M, and the amino acid modification at position T394 is T394W, and wherein the numbering of amino acid residues is according to the EU index as set forth in Kabat, more particularly, the Fc1 comprises amino acid modifications T350V, L351Y, F405A and Y407V, and the Fc2 comprises amino acid modifications T350V, T366L, K392L and T394W.

10 . The multispecific antibody or antigen binding fragment thereof of claim 7 , wherein at least one of the Fc1 and Fc2 comprises one or more mutations that enhance binding of the multispecific antibody or antigen binding fragment thereof to the neonatal Fc receptor (FcRn), preferably the one or more mutations enhance the binding at an acidic pH, more preferably the at least one of the Fc1 and Fc2 has the M252Y/S254T/T256E (YTE) mutations, wherein the numbering of amino acid residues is according to the EU index as set forth in Kabat.

11 . The multispecific antibody or antigen binding fragment thereof of claim 7 , wherein at least one of the Fc1 and Fc2 comprises one or more mutations that reduce or eliminate the effector function, preferably the at least one of the Fc1 and Fc2 has one or more amino acid modifications at positions L234, L235, D270, N297, E318, K320, K322, P331, and P329, such as one, two or three mutations of L234A, L235A and P331S, wherein the numbering of amino acid residues is according to the EU index as set forth in Kabat.

12 . A multispecific antibody or antigen binding fragment thereof comprising a first heavy chain comprising an amino acid sequence at least 90% identical to SEQ ID NO:24, and a first light chain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 25, and a second heavy chain comprising an amino acid sequence at least 90% identical to SEQ ID NO:26.

13 . The multispecific antibody or antigen binding fragment thereof of claim 12 , wherein the first heavy chain comprises the amino acid sequence of SEQ ID NO:24, the first light chain comprises the amino acid sequence of SEQ ID NO:25, and the second heavy chain comprises the amino acid sequence of SEQ ID NO:26.

14 . An isolated nucleic acid sequence encoding the multispecific antibody or antigen binding fragment thereof of claim 1 .

15 . A vector comprising the isolated nucleic acid of claim 14 .

16 . A host cell comprising the isolated nucleic acid of claim 14 or the vector of claim 15 .

17 . A method of producing a multispecific antibody or antigen binding fragment thereof, the method comprising culturing the host cell of claim 16 under conditions to produce the multispecific antibody or antigen binding fragment thereof and recovering the multispecific antibody or antigen binding fragment thereof.

18 . A pharmaceutical composition comprising the multispecific antibody or antigen binding fragment thereof of claim 1 and a pharmaceutically acceptable carrier.

19 . A method of treating or detecting a disorder, preferably a neurological disorder, in a subject in need thereof, comprising administering to the subject the multispecific antibody or antigen-binding fragment of claim 1 , prefer

Assignees

• Janssen Pharmaceutica Nv

Inventors

• Singh Sanjaya
• Yang Danlin
• Dasgupta Bidisha
• Nanjunda Rupesh
• Zwolak Adam
• Edwards Wilson
• Del Rosario Brian
• Kelly Thomas

Classifications

• C07K2317/92

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

• C07K2317/31

  multispecific · CPC title

• C07K16/2881

  against CD71 · CPC title

• A61K2039/505

  comprising antibodies · CPC title

• C07K2317/622

  Single chain antibody (scFv) · CPC title