Granted Patent
B2
US 12,331,132 · App. 18/206,354 · Granted Jun 17, 2025
Multispecific polypeptide constructs having constrained CD3 binding and related methods and uses
Inventors:
Brendan P. Eckelman (La Jolla, CA); Michael D. Kaplan (La Jolla, CA); Katelyn M. Willis (La Jolla, CA); John C. Timmer (La Jolla, CA)
Assignee:
Inhibrx Biosciences, Inc.
C07K16/30A61P35/00C07K1/14C07K2317/31C07K2317/522C07K2317/55C07K2317/569C07K2317/622C07K2317/624C07K2317/64C07K2317/66C07K2317/732C07K2317/734C07K2317/75C07K2317/92
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Abstract
The invention relates generally to multispecific polypeptides having constrained CD3 binding. In some embodiments, components of the multispecific polypeptides are connected by a non-cleavable linker. Also provided are methods of making and using these multispecific polypeptides in a variety of therapeutic, diagnostic and prophylactic indications.
Claims (35)
1. A multispecific polypeptide construct, the multispecific polypeptide construct comprising a first component comprising an immunoglobulin Fc region and a second component comprising a CD3-binding region, wherein:
the CD3 binding region is an anti-CD3 disulphide-stabilized Fv antibody fragment (dsFv) comprising a variable heavy chain region (VH) and a variable light chain region (VL);
the Fc region is a heterodimeric Fc comprising a first Fc polypeptide and a second Fc polypeptide and the VH and VL of the anti-CD3 antibody or antigen binding fragment are linked to opposite polypeptides of the heterodimeric Fc;
the first and second components are coupled by a non-cleavable linker, wherein the Fc region is linked to the N-terminus of the CD3-binding region;
the first component comprises at least one antigen binding domain that bind a tumor associated antigen (TAA), wherein the at least one antigen binding domain is linked to the N-terminus of the Fc region and wherein the at least one antigen binding domain is an sdAb.
2. The multispecific polypeptide construct of claim 1 , wherein the at least one antigen binding domain is a first antigen binding domain and a second antigen binding domain, wherein each of the at least one antigen binding domain of the first component is linked amino-terminal to the Fc region.
3. The multispecific polypeptide construct of claim 1 , wherein the CD3-binding region binds CD3.
4. The multispecific polypeptide construct of claim 1 , wherein each of the Fc polypeptides of the heterodimeric Fc independently comprise at least one amino acid modification.
5. The multispecific polypeptide construct of claim 4 , wherein the at least one modification is selected from a steric modification(s), a knob-into-hole modification(s), a charge mutation(s) to increase electrostatic complementarity of the polypeptides, a modification(s) to alter the isoelectric point, or combinations thereof.
6. The multispecific polypeptide construct of claim 1 , wherein the non-cleavable linker is a polypeptide linker.
7. The multispecific polypeptide construct of claim 1 , wherein the non-cleavable linker is a polypeptide that is 3 to 18 amino acids in length.
8. The multispecific polypeptide construct of claim 1 , wherein the non-cleavable linker comprises (GGS) n, wherein n is 1 to 10;
(GGGGS)n (SEQ ID NO: 173), wherein n is 1 to 10; or
(GGGGGS)n (SEQ ID NO:172), wherein n is 1 to 4.
9. The multispecific polypeptide construct of claim 1 , wherein the non-cleavable linker is or comprises an amino acid sequence selected from the group consisting of GGS; GGGGS (SEQ ID NO: 149); GGGGGS (SEQ ID NO: 135); (GGS) 2 (SEQ ID NO: 10); GGSGGSGGS (SEQ ID NO: 11); GGSGGSGGSGGS (SEQ ID NO: 12); GGSGGSGGSGGSGGS (SEQ ID NO: 13); GGGGGSGGGGGSGGGGGS (SEQ ID NO: 119); GGSGGGGSGGGGSGGGGS (SEQ ID NO: 147); and GGGGSGGGGSGGGGS (SEQ ID NO: 170).
10. The multispecific polypeptide construct of claim 1 , wherein the multispecific polypeptide construct comprises at least (i) a first polypeptide comprising the first Fc polypeptide of the heterodimeric Fc region, the linker and the VH domain of the anti-CD3 antibody or antigen binding fragment thereof; and (ii) a second polypeptide comprising the second Fc polypeptide of the heterodimeric Fc region, the linker and the VL domain of the anti-CD3 antibody or antigen binding fragment thereof, wherein one or both of the first and second polypeptide comprise the at least one antigen-binding domain that binds to a tumor associated antigen (TAA).
11. The multispecific polypeptide construct of claim 10 , wherein the VH of the anti-CD3 antibody or antigen-binding fragment is on the same polypeptide as the at least one antigen-binding domain that binds to a tumor associated antigen (TAA).
12. The multispecific polypeptide construct of claim 10 , wherein the polypeptide comprising the VL of the anti-CD3 antibody or antigen-binding fragment does not contain the at least one antigen-binding domain that binds to a tumor associated antigen (TAA).
13. The multispecific polypeptide construct of claim 10 , wherein only one of the first and second polypeptide comprises the at least one antigen-binding domain that binds a TAA.
14. The multispecific polypeptide construct of claim 1 , wherein the multispecific polypeptide construct comprises:
at least a first antigen binding domain and a second antigen binding domain, wherein the first antigen binding domain and second antigen binding domain bind to the same TAA;
at least a first antigen binding domain and a second antigen binding domain wherein the first antigen binding domain and the second antigen binding domain binds different epitopes of the same TAA;
at least a first antigen binding domain and a second antigen binding domain wherein the first antigen binding domain and the second antigen binding domain bind the same epitope of the same TAA; or
at least a first antigen binding domain and a second antigen binding domain wherein the first antigen binding domain and the second antigen binding domain bind a different TAA.
15. The multispecific polypeptide construct of claim 1 , wherein:
the anti-CD3 antibody or antigen-binding fragment thereof comprises a VH CDR1 comprising the amino acid sequence GFTFNTYAMN (SEQ ID NO: 211); a VH CDR2 comprising the amino acid sequence RIRSKYNNYATY (SEQ ID NO: 212); a VH CDR3 comprising the amino acid sequence HGNFGNSYVSWFAY (SEQ ID NO: 18), a VL CDR1 comprising the amino acid sequence RSSTGAVTTSNYAN (SEQ ID NO: 19); a VL CDR2 comprising the amino acid sequence GTNKRAP (SEQ ID NO: 20); and a VL CDR3 comprising the amino acid sequence ALWYSNLWV (SEQ ID NO: 21).
16. The multispecific polypeptide of claim 1 , wherein:
the anti-CD3 antibody or antigen-binding fragment comprises a VH CDR1 sequence that includes at least the amino acid sequence GFTFNTYAMN (SEQ ID NO: 211); a VH CDR2 sequence that includes at least the amino acid sequence RIRSKYNNYATY (SEQ ID NO: 212); a VH CDR3 sequence that includes at least the amino acid sequence HGNFGNSYVSWFAY (SEQ ID NO: 18), a VL CDR1 sequence that includes at least the amino acid sequence GSSTGAVTTSNYAN (SEQ ID NO: 229); a VL CDR2 sequence that includes at least the amino acid sequence GTNKRAP (SEQ ID NO: 230); and a VL CDR3 sequence that includes at least the amino acid sequence ALWYSNHWV (SEQ ID NO: 225).
17. The multispecific polypeptide construct of claim 16 , wherein the anti-CD3 dsFv comprises:
a VH having the amino acid sequence of any of SEQ ID NOS: 44, 50, 53-56, and 60-62 or a sequence that exhibits at least 90%, sequence identity to any of SEQ ID NOS: 44, 50, 53-56, and 60-62; and
a VL having the amino acid sequence of any of SEQ ID NOS: 64, 72, 80, 81, 191, and 200 or a sequence that exhibits at least 90% sequence identity to any of SEQ ID NOS: 64, 72, 80, 81, 191, and 200.
18. The multispecific polypeptide construct of claim 16 , wherein the anti-CD3 dsFv comprises a VH having a sequence that exhibits at least 90% sequence identity to SEQ ID NO: 44; and a VL having a sequence that exhibits at least 90% sequence identity SEQ ID NO: 72.
19. The multispecific polypeptide construct of claim 1 , wherein the anti-CD3 dsFv comprises the amino acid sequence of SEQ ID NO: 44 and the amino acid sequence of SEQ ID NO: 72.
20. The multispecific polypeptide construct of claim 1 , wherein the multispecific polypeptide construct is conjugated to an agent selected from the group consisting of the following: a therapeutic agent, an antineoplastic agent, a toxin or fragment thereof, a detectable moiety, and a diagnostic agent.
21. A pharmaceutical composition comprising the multispecific polypeptide construct of claim 1 and a pharmaceutically acceptable carrier.
Assignments (3)
SECURITY INTEREST
Recorded Jan 14, 2025
From: INHIBRX BIOSCIENCES, INC.
To: OXFORD FINANCE LLC; OXFORD FINANCE LLC
Reel/Frame 069894/0045 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jun 10, 2024
From: INHIBRX, INC.
To: INHIBRX BIOSCIENCES, INC.
Reel/Frame 067679/0635 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jul 5, 2023
From: ECKELMAN, BRENDAN P.; KAPLAN, MICHAEL D.; WILLIS, KATELYN M.; TIMMER, JOHN C.
To: INHIBRX, INC.
Reel/Frame 064156/0732 →
Continuity (3)
Provisional Application
62656331
· Apr 11, 2018
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