IP Library › Granted Patent US 12,371,496
Granted Patent B2
US 12,371,496 · App. 17/246,771 · Granted Jul 29, 2025

T cell recruiting polypeptides based on CD3 reactivity

Inventors: Annelies Roobrouck (Oudenaarde, BE); Diane Van Hoorick (Laarne, BE); João Vieira (Didcot, BE)
Assignee: Ablynx N.V.
C07K16/2809A61P35/00C07K16/18C07K16/2863C07K16/2887C07K16/30C07K16/32A61K2039/505C07K2317/14C07K2317/31C07K2317/565C07K2317/567C07K2317/569C07K2317/73C07K2317/76C07K2317/92C07K2317/94
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Quick Facts
Patent No.
US 12,371,496
App. No.
17/246,771
Granted
Jul 29, 2025
Kind
B2
Abstract

T cell recruiting polypeptides are provided that bind CD3 on a T cell. The polypeptides can be used in methods for treatment of cancers.

Claims (48)

1. A polypeptide comprising a first and a second immunoglobulin single variable domain (ISV), wherein each of the first ISV and the second ISV is a Nanobody, a V HH , or a camelized V H , and wherein:

said first ISV binds to cluster of differentiation 3 (CD3) present on a T cell;

said second ISV binds to a first antigen on a target cell;

wherein said first antigen is different from said CD3;

wherein said target cell is different from said T cell; and

wherein said first ISV consists of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively), in which:

CDR1 comprises the amino acid sequence of SEQ ID NO: 94, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 134;

CDR1 comprises the amino acid sequence of SEQ ID NO: 90, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 135;

CDR1 comprises the amino acid sequence of SEQ ID NO: 95, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 135;

CDR1 comprises the amino acid sequence of SEQ ID NO: 96, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 136;

CDR1 comprises the amino acid sequence of SEQ ID NO: 97, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 137;

CDR1 comprises the amino acid sequence of SEQ ID NO: 98, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 137;

CDR1 comprises the amino acid sequence of SEQ ID NO: 99, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 138;

CDR1 comprises the amino acid sequence of SEQ ID NO: 99, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 139;

CDR1 comprises the amino acid sequence of SEQ ID NO: 95, CDR2 comprises the amino acid sequence of SEQ ID NO: 119, and CDR3 comprises the amino acid sequence of SEQ ID NO: 140;

CDR1 comprises the amino acid sequence of SEQ ID NO: 100, CDR2 comprises the amino acid sequence of SEQ ID NO: 120, and CDR3 comprises the amino acid sequence of SEQ ID NO: 141;

CDR1 comprises the amino acid sequence of SEQ ID NO: 100, CDR2 comprises the amino acid sequence of SEQ ID NO: 121, and CDR3 comprises the amino acid sequence of SEQ ID NO: 142; or

CDR1 comprises the amino acid sequence of SEQ ID NO: 100, CDR2 comprises the amino acid sequence of SEQ ID NO: 122, and CDR3 comprises the amino acid sequence of SEQ ID NO: 143.

2. The polypeptide according to claim 1 , wherein said first antigen is a tumor antigen.

3. The polypeptide according to claim 1 , further comprising a third ISV, which binds to a second antigen on a target cell, wherein said second antigen is different from said first antigen, wherein the third ISV is a Nanobody, a V HH , or a camelized V H .

4. The polypeptide according to claim 3 , wherein said second antigen is a tumor antigen.

5. The polypeptide according to claim 1 , further comprising a serum protein binding moiety, wherein the serum binding moiety is an ISV that binds serum albumin, wherein the ISV that binds serum albumin is a Nanobody, a V HH , or a camelized V H .

6. The polypeptide according to claim 5 , wherein the ISV that binds serum albumin is a humanized V HH .

7. A polypeptide that comprises an immunoglobulin single variable (ISV) domain that specifically binds CD3, wherein the ISV is a Nanobody, a V HH , or a camelized V H that comprises or consists of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively), in which:

CDR1 comprises the amino acid sequence of SEQ ID NO: 94, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 134;

CDR1 comprises the amino acid sequence of SEQ ID NO: 90, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 135;

CDR1 comprises the amino acid sequence of SEQ ID NO: 95, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 135;

CDR1 comprises the amino acid sequence of SEQ ID NO: 96, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 136;

CDR1 comprises the amino acid sequence of SEQ ID NO: 97, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 137;

CDR1 comprises the amino acid sequence of SEQ ID NO: 98, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 137;

CDR1 comprises the amino acid sequence of SEQ ID NO: 99, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 138;

CDR1 comprises the amino acid sequence of SEQ ID NO: 99, CDR2 comprises the amino acid sequence of SEQ ID NO: 118, and CDR3 comprises the amino acid sequence of SEQ ID NO: 139;

CDR1 comprises the amino acid sequence of SEQ ID NO: 95, CDR2 comprises the amino acid sequence of SEQ ID NO: 119, and CDR3 comprises the amino acid sequence of SEQ ID NO: 140;

CDR1 comprises the amino acid sequence of SEQ ID NO: 100, CDR2 comprises the amino acid sequence of SEQ ID NO: 120, and CDR3 comprises the amino acid sequence of SEQ ID NO: 141;

CDR1 comprises the amino acid sequence of SEQ ID NO: 100, CDR2 comprises the amino acid sequence of SEQ ID NO: 121, and CDR3 comprises the amino acid sequence of SEQ ID NO: 142; or

CDR1 comprises the amino acid sequence of SEQ ID NO: 100, CDR2 comprises the amino acid sequence of SEQ ID NO: 122, and CDR3 comprises the amino acid sequence of SEQ ID NO: 143.

8. The polypeptide according to claim 7 , further comprising a serum protein binding moiety, wherein the serum binding moiety is an ISV that binds serum albumin, wherein the ISV that binds serum albumin is a Nanobody, a V HH , or a camelized V H .

9. The polypeptide according to claim 8 , wherein the ISV that binds serum albumin is a humanized V HH .

10. A pharmaceutical composition comprising a polypeptide according to claim 1 .

11. The polypeptide according to claim 1 , wherein each of the first ISV and the second ISV is a humanized V HH .

12. The polypeptide according to claim 1 , wherein the first ISV consists of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively), in which: CDR1 comprises the amino acid sequence of SEQ ID NO: 94; CDR2 comprises the amino acid sequence of SEQ ID NO: 118; and CDR3 comprises the amino acid sequence of SEQ ID NO: 134.

13. The polypeptide according to claim 1 , wherein the first ISV is selected from the group consisting of SEQ ID NOs: 66-80.

14. The polypeptide according to claim 2 , wherein the first antigen is a tumor-associated antigen.

15. The polypeptide according to claim 4 , wherein the second antigen is a tumor-associated antigen.

16. The polypeptide according to claim 3 , wherein the third ISV is a humanized V HH .

17. The polypeptide according to claim 7 , wherein the ISV is a humanized V HH .

18. The polypeptide according to claim 7 , wherein the ISV consists of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3, respectively), in which: CDR1 comprises the amino acid sequence of SEQ ID NO: 94; CDR2 comprises the amino acid sequence of SEQ ID NO: 118; and CDR3 comprises the amino acid sequence of SEQ ID NO: 134.

19. The polypeptide according to 7 , wherein the polypeptide is selected from the group consisting of SEQ ID NOs: 66-80.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 22, 2022
From: ROOBROUCK, ANNELIES; VAN HOORICK, DIANE; VIEIRA, JOÃO
To: ABLYNX N.V.
Reel/Frame 061175/0727 →
Continuity (3)
Continuation 15573298
Provisional Application 62160794 · May 13, 2015
Related Publication 20210363251A1 · Nov 25, 2021
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