IP Library › Granted Patent US 12,325,733

Granted Patent B2

US 12,325,733 · App. 17/290,188 · Granted Jun 10, 2025

Binding domain

Inventors: Anna Bulek (London, GB); Martin Pulé (London, GB); Shaun Cordoba (London, GB); Simon Thomas (London, GB); Shimobi Onuoha (London, GB); Mathieu Ferrari (London, GB); Vania Baldan (London, GB)

Assignee: AUTOLUS LIMITED

C07K14/7051 A61K47/6849 C07K14/70517 C07K16/2809 G01N33/57407 G01N33/57426 C07K2317/56 C07K2319/03

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Quick Facts

Patent No.: US 12,325,733
App. No.: 17/290,188
Granted: Jun 10, 2025
Kind: B2

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Abstract

The present invention provides a variant antigen-binding domain which comprises at least one mutation in the VH domain compared to a reference antibody and which displays an increased affinity for TRBC2 over the reference antibody. It further provides an antibody, a chimeric antigen receptor (CAR), and a bispecific T-cell engager (BiTE), a cell which comprises said CAR, and a conjugate comprising said variant antigen-binding domain or said antibody. Additionally, it provides medical uses, diagnostic methods and methods of personalised medicine that exploit the products of the invention.

Claims (137)

1. A variant antigen-binding domain which comprises mutations in the VH domain compared to a reference antibody having a VH domain with the sequence shown in SEQ ID NO: 1 and a VL domain with the sequence shown in SEQ ID NO: 2, wherein the variant antigen-binding domain comprises the following mutation combinations:

T28K, Y32F, A100N, Y102F in the VH domain;

T28K, Y32F, A100N, Y102L, N103M in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, Y102L, N103W in the VH domain and N35K in the VL domain;

T28K, Y32F, A100N, Y102L, N103W in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, N103E in the VH domain;

V2K, T28K, Y32F, A100N in the VH domain;

N103E, T28K, T32F, A100N in the VH domain and N35M in the VL domain;

T28K, Y32F, A100N, N103F in the VH domain and N35K in the VL domain;

T28K, Y32F, A100N, A107S in the VH domain;

G31S, T28K, Y32F, A100N in the VH domain;

V2R, T28K, Y32F, A100N in the VH domain;

T28K, Y32F, A100N, N103M in the VH domain and N35R in the VL domain;

N103W, T28K, Y32F, A100N in the VH domain and N35F in the VL domain;

T28K, G31R, Y32F, A100N in the VH domain;

N103F, T28K, T32F, A100N in the VH domain and N35F in the VL domain;

N103F, T28K, T32F, A100N in the VH domain and N35M in the VL domain;

N103M, T28K, T32F, A100N in the VH domain and N32F in the VL domain;

T28K, Y32F, A100N, N103S in the VH domain;

T28K, Y32F, A100N, N103Q in the VH domain;

Y27F, T28K, Y32F, A100N in the VH domain;

N103F, T28K, Y32F, A100N in the VH domain and N35Y in the VL domain;

N103W, T28K, Y32F, A100N in the VH domain and N35M in the VL domain;

N103L, T28K, Y32F, A100N in the VH domain and N35F in the VL domain;

N103L, T28K, Y32F, A100N in the VH domain and N35K in the VL domain;

T28K, Y32F, A100N, N103W in the VH domain;

T28K, Y32F, A100N, N103F in the VH domain;

N103L, T28K, Y32F, A100N in the VH domain and N35Y in the VL domain;

N103W, T28K, Y32F, A100N in the VH domain and N35K in the VL domain;

N103L, T28K, Y32F, A100N in the VH domain and N35R in the VL domain;

N103L, T28K, Y32F, A100N in the VH domain and N35M in the VL domain;

N103S, T28K, Y32F, A100N in the VH domain and N35Y in the VL domain;

N103S, T28K, Y32F, A100N in the VH domain and N35F in the VL domain;

T28K, Y32F, A100N, N103L in the VH domain;

T28K, Y32F, A100N, N103M in the VH domain and N35Y in the VL domain;

N103S, T28K, Y32F, A100N in the VH domain and N35R in the VL domain;

T28K, Y32F, R98K, A100N in the VH domain;

N103S, T28K, Y32F, A100N in the VH domain and N35K in the VL domain;

T28K, Y32F, A100N in the VH domain and N35F in the VL domain;

N103W, T28K, T32F, A100N in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, N103M in the VH domain;

N103S, T28K, Y32F, A100N in the VH domain and N35M in the VL domain;

T28K, Y32F, A100N in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, N103Y in the VH domain;

T28K, Y32F, A100N, N103A in the VH domain;

T28K, Y32F, A100N, N103H in the VH domain;

T28K, Y32F, A100N, N103M in the VH domain and N35M in the VL domain;

T28K, Y32F, A100N in the VH domain and N35K in the VL domain;

T28K, Y32F, A100N in the VH domain and R55K in the VL domain;

T28K, Y32F, A100N in the VH domain;

Y27W, T28K, Y32F, A100N in the VH domain;

T28K, Y32F, A100N, Y102F in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, Y102F, N103M in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, Y102F, N103M in the VH domain and N35F in the VL domain;

Y27M, T28K, Y32F, A100N in the VH domain;

T28K, G31K, Y32F, A100N in the VH domain;

T28K, Y32F, A100N, Y102F, N103M in the VH domain and N35K in the VL domain; or

Y27N, T28K, V32F, A100N in the VH domain;

and wherein the variant antigen-binding domain displays an increased affinity for TRBC2 over the reference antibody.

2. An antibody comprising a variant antigen-binding domain according to claim 1 .

3. A chimeric antigen receptor (CAR) comprising a variant antigen-binding domain according to claim 1 , a spacer, a transmembrane domain and an endodomain.

4. A bispecific T-cell engager comprising a variant antigen-binding domain according to claim 1 and a T-cell activation domain.

5. A nucleic acid sequence encoding:

a variant antigen-binding domain, an antibody comprising a variant antigen-binding domain, a CAR comprising a variant antigen-binding domain, or a bispecific T-cell engager comprising a variant antigen-binding domain,

wherein the variant binding domain comprises mutations in the VH domain compared to a reference antibody having a VH domain with the sequence shown in SEQ ID NO: 1 and a VL domain with the sequence shown in SEQ ID NO: 2,

wherein the mutations comprise the mutation combinations:

T28K, Y32F, A100N, Y102F in the VH domain;

T28K, Y32F, A100N, Y102L, N103M in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, Y102L, N103W in the VH domain and N35K in the VL domain;

T28K, Y32F, A100N, Y102L, N103W in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, N103E in the VH domain;

V2K, T28K, Y32F, A100N in the VH domain;

N103E, T28K, T32F, A100N in the VH domain and N35M in the VL domain;

T28K, Y32F, A100N, N103F in the VH domain and N35K in the VL domain;

T28K, Y32F, A100N, A107S in the VH domain;

G31S, T28K, Y32F, A100N in the VH domain;

V2R, T28K, Y32F, A100N in the VH domain;

T28K, Y32F, A100N, N103M in the VH domain and N35R in the VL domain;

N103W, T28K, Y32F, A100N in the VH domain and N35F in the VL domain;

T28K, G31R, Y32F, A100N in the VH domain;

N103F, T28K, T32F, A100N in the VH domain and N35F in the VL domain;

N103F, T28K, T32F, A100N in the VH domain and N35M in the VL domain;

N103M, T28K, T32F, A100N in the VH domain and N32F in the VL domain;

T28K, Y32F, A100N, N103S in the VH domain;

T28K, Y32F, A100N, N103Q in the VH domain;

Y27F, T28K, Y32F, A100N in the VH domain;

N103F, T28K, Y32F, A100N in the VH domain and N35Y in the VL domain;

N103W, T28K, Y32F, A100N in the VH domain and N35M in the VL domain;

N103L, T28K, Y32F, A100N in the VH domain and N35F in the VL domain;

N103L, T28K, Y32F, A100N in the VH domain and N35K in the VL domain;

T28K, Y32F, A100N, N103W in the VH domain;

T28K, Y32F, A100N, N103F in the VH domain;

N103L, T28K, Y32F, A100N in the VH domain and N35Y in the VL domain;

N103W, T28K, Y32F, A100N in the VH domain and N35K in the VL domain;

N103L, T28K, Y32F, A100N in the VH domain and N35R in the VL domain;

N103L, T28K, Y32F, A100N in the VH domain and N35M in the VL domain;

N103S, T28K, Y32F, A100N in the VH domain and N35Y in the VL domain;

N103S, T28K, Y32F, A100N in the VH domain and N35F in the VL domain;

T28K, Y32F, A100N, N103L in the VH domain;

T28K, Y32F, A100N, N103M in the VH domain and N35Y in the VL domain;

N103S, T28K, Y32F, A100N in the VH domain and N35R in the VL domain;

T28K, Y32F, R98K, A100N in the VH domain;

N103S, T28K, Y32F, A100N in the VH domain and N35K in the VL domain;

T28K, Y32F, A100N in the VH domain and N35F in the VL domain;

N103W, T28K, T32F, A100N in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, N103M in the VH domain;

N103S, T28K, Y32F, A100N in the VH domain and N35M in the VL domain;

T28K, Y32F, A100N in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, N103Y in the VH domain;

T28K, Y32F, A100N, N103A in the VH domain;

T28K, Y32F, A100N, N103H in the VH domain;

T28K, Y32F, A100N, N103M in the VH domain and N35M in the VL domain;

T28K, Y32F, A100N in the VH domain and N35K in the VL domain;

T28K, Y32F, A100N in the VH domain and R55K in the VL domain;

T28K, Y32F, A100N in the VH domain;

Y27W, T28K, Y32F, A100N in the VH domain;

T28K, Y32F, A100N, Y102F in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, Y102F, N103M in the VH domain and N35R in the VL domain;

T28K, Y32F, A100N, Y102F, N103M in the VH domain and N35F in the VL domain;

Y27M, T28K, Y32F, A100N in the VH domain;

T28K, G31K, Y32F, A100N in the VH domain;

T28K, Y32F, A100N, Y102F, N103M in the VH domain and N35K in the VL domain; or

Y27N, T28K, V32F, A100N in the VH domain; and

wherein the variant antigen-binding domain displays an increased affinity for TRBC2 over the reference antibody.

6. A vector comprising a nucleic acid sequence according to claim 5 .

7. A cell which comprises a CAR according to claim 3 .

8. A method for making a cell, which comprises the step of transducing or transfecting a cell with a vector according to claim 6 .

9. A conjugate comprising an antibody comprising a variant antigen-binding domain according to claim 1 , and a detectable entity or a chemotherapeutic entity.

10. The conjugate according to claim 9 , which comprises a chemotherapeutic entity.

11. A method for treating a T-cell lymphoma or leukaemia in a subject which comprises the step of administering:

a cell according to claim 7 , or

an antibody comprising a variant antigen-binding domain according to claim 1 , a bispecific T-cell engager comprising a variant antigen-binding domain according to claim 1 or a conjugate comprising a variant antigen-binding domain according to claim 1 to a subject,

wherein the malignant T-cells express TRBC2.

12. A method for diagnosing a T-cell lymphoma or leukaemia in a subject which comprises the step of contacting a variant antigen-binding domain according to claim 1 , or an antibody comprising the variant antigen-binding domain, to a sample comprising T-cells from the subject and determining the percentage of TRBC2 positive T-cells in the sample, wherein T-cell lymphoma or leukaemia is diagnosed when the percentage is 70%, or 75%, or 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or more.

13. A method for identifying subjects with a T-cell lymphoma or leukaemia eligible for treatment with a cell comprising a CAR comprising an antigen-binding domain according to claim 1 , or an antibody comprising a variant antigen-binding domain according to claim 1 , a bispecific T-cell engager comprising a variant antigen-binding domain according to claim 1 , or a conjugate comprising a variant antigen-binding domain according to claim 1 , comprising determining the percentage of TRBC2 positive T-cells in a sample comprising T-cells from the subject, wherein patients with a T-cell lymphoma or leukaemia are identified when the percentage is 70%, or 75%, or 80%, or 85%, or 90%, or 95%, or 96%, or 97%, or 98%, or 99%, or more.

14. A method for selecting a therapy comprising a cell comprising a CAR comprising an antigen-binding domain according to claim 1 , or an antibody comprising a variant antigen-binding domain according to claim 1 , a bispecific T-cell engager comprising a variant antigen-binding domain according to claim 1 , or a conjugate comprising a variant antigen-binding domain according to claim 1 , for the treatment of a subject, comprising determining the percentage of TRBC2 positive T-cells in a sample comprising T-cells from the subject.

15. The method according to claim 11 , wherein the T-cell lymphoma or leukaemia is selected from peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS); angio-immunoblastic T-cell lymphoma (AITL), anaplastic large cell lymphoma (ALCL), enteropathy-associated T-cell lymphoma (EATL), hepatosplenic T-cell lymphoma (HSTL), extranodal NK/T-cell lymphoma nasal type, cutaneous T-cell lymphoma, primary cutaneous ALCL, T cell prolymphocytic leukaemia and T-cell acute lymphoblastic leukaemia.

Assignments (2)

PATENT SECURITY AGREEMENT Recorded Jul 30, 2026

From: AUTOLUS LIMITED

To: PERCEPTIVE CREDIT HOLDINGS V, LP, AS ADMINISTRATIVE AGENT

Reel/Frame 076084/0283 →

ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 5, 2021

From: BULEK, ANNA; PULÉ, MARTIN; CORDOBA, SHAUN; THOMAS, SIMON; ONUOHA, SHIMOBI; FERRARI, MATHIEU; BALDAN, VANIA

To: AUTOLUS LIMITED

Reel/Frame 056137/0410 →

Priority Claims (1)

GB 1817822 · Oct 31, 2018 · national

Continuity (1)

Related Publication 20220041718A1 · Feb 10, 2022

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