JPWO2020257871A5

JPWO2020257871A5 -

Info

Publication number: JPWO2020257871A5
Application number: JP2021577332A
Authority: JP
Publication date: 2023-07-04

Claims

a BTN2A1 antagonist,
(A) to inhibit activation of γδ T cells expressing Vγ9+ TCR in a subject;
(B) to suppress or inhibit a Vγ9+γδT cell response in a subject;
wherein said BTN2A1 antagonist is:
i) inhibit the formation of BTN2A1/BTN3A1 complexes on the surface of cells;
ii) inhibits BTN2A1 binding to Vγ9;
iii) inhibit the binding of the BTN2A1/BTN3A1 complex to the Vγ9+ TCR; and/or
iv) decrease the activity and/or survival of cells expressing BTN2A1; or
(C) to prevent, treat, slow progression, prevent recurrence of, or ameliorate symptoms of autoimmune disease, transplant rejection, or graft-versus-host disease in a subject in need thereof;
BTN2A1 antagonist of.

A BTN2A1 antagonist for inhibiting the activation of γδ T cells expressing Vγ9+ TCR in vitro or in vitro, comprising:
culturing the γδ T cells and cells expressing BTN2A1 in the presence of a BTN2A1 antagonist;
wherein said BTN2A1 antagonist is:
i) the formation of a BTN2A1/BTN3A1 complex on the surface of the cell;
ii) BTN2A1 binds to Vγ9; and/or
iii) binding of the BTN2A1/BTN3A1 complex to the Vγ9+ TCR;
A BTN2A1 antagonist that inhibits

specifically binds to BTN2A1;
i) the formation of a BTN2A1/BTN3A1 complex on the surface of the cell;
ii) BTN2A1 binds to Vγ9; and/or
iii) binding of the BTN2A1/BTN3A1 complex to the Vγ9+ TCR;
A BTN2A1 antagonist that inhibits

1(A), 2 and 3, wherein the BTN2A1 antagonist inhibits activation of Vγ9Vδ2+γδ T cells and/or inhibits activation of Vγ9Vδ2−γδ T cells. BTN2A1 antagonist.

The BTN2A1 antagonist of claim 1(B), wherein said BTN2A1 antagonist suppresses or inhibits Vγ9+γδ T cell responses and/or suppresses or inhibits Vγ9Vδ2-γδ T cell responses.

The BTN2A1 antagonist of any one of claims 1-5, wherein said BTN2A1/BTN3A1 complex comprises one or more additional molecules.

7. The BTN2A1 antagonist of claim 6, wherein said BTN2A1/BTN3A1 complex comprises BTN3A2 and/or BTN3A3.

wherein the BTN2A1 antagonist is
i) suppresses or inhibits one or more of cytolytic function, cytokine production of one or more cytokines, or proliferation of said γδT cells; and/or
ii) inhibits phosphoantigen-mediated activation of γδ T cells; and/or
iii) inhibit the association of BTN2A1 and BTN3A1; and/or
iv) The BTN2A1 antagonist of any one of claims 1-7, which inhibits binding of BTN2A1 to the germline coding region of Vγ9 and/or distal to the delta chain.

9. The BTN2A1 antagonist of claim 8, wherein said BTN2A1 antagonist inhibits direct association of BTN2A1 and BTN3A1.

10. The BTN2A1 antagonist of any one of claims 1-9, wherein the BTN2A1 antagonist inhibits the binding of the BTN2A1/BTN3A1 complex to the germline coding region of Vδ2 (such as the CDR2 loop and/or the CDR3 loop of the TCRγ chain). BTN2A1 antagonist.

11. Any one of claims 1-10, wherein said BTN2A1 antagonist alters one or more of the extracellular domains (IgV and/or IgC) of the BTN2A1 molecule to switch this BTN2A1 molecule from stimulatory BTN2A1 to non-stimulatory BTN2A1. A BTN2A1 antagonist according to any one of the preceding paragraphs.

12. The BTN2A1 antagonist of claim 11, wherein said BTN2A1 antagonist alters one or more of the extracellular domains (IgV and/or IgC) of said BTN2A1 molecule and inhibits phosphoantigen activation.

a BTN2A1 agonist,
(A) to activate γδ T cells expressing the Vγ9+ TCR in a subject;
(B) to induce or enhance a Vγ9+γδ T cell response in a subject;
wherein said BTN2A1 agonist is:
i) promoting the formation of BTN2A1/BTN3A1 complexes on the surface of cells;
ii) induce Vγ9+ TCR ligation on the surface of γδ T cells; and/or
iii) increases the activity and/or survival of cells expressing BTN2A1;
BTN2A1 agonists of.

A BTN2A1 agonist for in vitro or ex vivo activation of γδ T cells expressing Vγ9+ TCR, comprising:
culturing the γδ T cells and cells expressing BTN2A1 in the presence of a BTN2A1 agonist;
wherein said BTN2A1 agonist:
i) promoting the formation of BTN2A1/BTN3A1 complexes on the surface of antigen-presenting cells;
ii) induce Vγ9+ TCR ligation on the surface of γδ T cells; and/or
iii) increases the activity and/or survival of cells expressing BTN2A1;
BTN2A1 agonist.

The BTN2A1 agonist is
15. The BTN2A1 agonist according to claim 13(A) or 14, which activates Vγ9Vδ2+γδ T cells and/or activates Vγ9Vδ2−γδ T cells.

The BTN2A1 agonist is
The BTN2A1 agonist of claim 13(B), which induces a Vγ9+γδT cell response and/or induces a Vγ9Vδ2-γδT cell response.

The BTN2A1 agonist of any one of claims 13-16, wherein said BTN2A1/BTN3A1 complex comprises one or more additional molecules.

18. The BTN2A1 agonist of claim 17, wherein said BTN2A1/BTN3A1 complex comprises BTN3A2 and/or BTN3A3.

The BTN2A1 agonist is
i) activates or induces one or more of cytolytic function, cytokine production of one or more cytokines, or proliferation of said γδT cells; and/or
ii) activates said γδ T cells independently of phosphoantigen binding; and/or
iii) promotes the association of BTN2A1 and BTN3A1; and/or
iv) bispecific for BTN2A1 and BTN3A1; and/or
v) altering one or more of the extracellular domains (IgV and/or IgC) of the BTN2A1 molecule to switch this BTN2A1 molecule from non-stimulatory BTN2A1 to stimulatory BTN2A1;
A BTN2A1 agonist according to any one of claims 13-18.

20. The BTN2A1 agonist of claim 19, wherein said BTN2A1 agonist promotes direct association of BTN2A1 and BTN3A1.
,

20. The BTN2A1 agonist of claim 19, wherein said BTN2A1 agonist cross-reacts with BTN3A1.

22. The BTN2A1 antagonist or agonist of any one of claims 1-21, wherein the BTN2A1 antagonist or agonist is a protein comprising an antigen binding domain.

the protein is
(i) a single-chain Fv fragment (scFv);
(ii) a dimeric scFv;
(iii) an Fv fragment;
(iv) single domain antibodies (sdAbs);
(v) Nanobodies;
(vi) diabodies, triabodies, tetrabodies, or higher multimers;
(vii) a Fab fragment;
(viii) a Fab'fragment;
(ix) an F(ab') fragment;
(x) F(ab') ₂ fragment;
(xi) any one of (i)-(x) linked to the Fc region of the antibody;
(xii) any one of (i)-(x) fused to an antibody or antigen-binding fragment thereof that binds to immune effector cells; or
(xiii) an antibody;
23. The BTN2A1 antagonist or agonist of claim 22, which is

The BTN2A1 antagonist of any one of claims 1-12, wherein said BTN2A1 antagonist is a soluble Vγ9+TCR.

25. The BTN2A1 antagonist of claim 24, wherein said soluble V[gamma]9+ TCR is monomeric or said soluble V[gamma]9+ TCR is multimeric.

wherein the BTN2A1 antagonist is
(i) an antibody comprising a heavy chain variable region (V _H ) comprising the sequence set forth in SEQ ID NO: 100 and a light chain variable region (V _L ) comprising the sequence set forth in SEQ ID NO: 101; or
(ii) an antibody comprising a heavy chain variable region (V _H ) comprising the sequence set forth in SEQ ID NO: 108 and a light chain variable region (V _L ) comprising the sequence set forth in SEQ ID NO: 109; or
(ii) an antibody comprising a heavy chain variable region (V _H ) comprising the sequence set forth in SEQ ID NO: 116 and a light chain variable region (V _L ) comprising the sequence set forth in SEQ ID NO: 117;
(iv) an antibody comprising a heavy chain variable region (V _H ) comprising the sequence set forth in SEQ ID NO: 124 and a light chain variable region (V _L ) comprising the sequence set forth in SEQ ID NO: 125;
(v) an antibody comprising a heavy chain variable region (V _H ) comprising the sequence set forth in SEQ ID NO: 132 and a light chain variable region (V _L ) comprising the sequence set forth in SEQ ID NO: 133;
The BTN2A1 antagonist according to any one of claims 1 to 12, which is

specifically binds to BTN2A1: and
(A)
(i) activates γδ T cells within a population of cells and/or increases the number of activated γδ T cells; and/or
(ii) increasing the proportion of γδ T cells expressing markers of γδ T cell activation; and/or
(iii) increase secretion of cytokines by γδ T cells; and/or
(iv) inducing γδ T cells to kill cancer cells and/or inhibit the proliferation of cancer cells and/or kill infected cells and/or inhibit the proliferation of infected cells and/or
(v) increasing the amount of markers of γδT cell activation expressed on the cell surface of γδT cells; or
(B)
(i) increasing the proportion of γδ T cells expressing CD25 on their cell surface; and/or
(ii) increase secretion of interferon-γ by γδ T cells; and/or
(iii) induce γδ T cells to kill cancer cells and/or inhibit the growth of cancer cells; and/or
(iv) increasing the amount of CD25 expressed on the cell surface of γδT cells;
BTN2A1 agonist.

(i) a light chain variable region (V _L ) comprising the sequence set forth in SEQ ID NO: 140 or its complementarity determining regions (CDRs) and a heavy chain variable region comprising the sequence set forth in SEQ ID NO: 144 or its CDRs region (V _H );
(ii) a V _L comprising the sequence set forth in SEQ ID NO: 148 or CDRs thereof and a V _H comprising the sequence set forth in SEQ ID NO: 152 or CDRs thereof;
(iii) V _L comprising the sequence set forth in SEQ ID NO: 156 or CDRs thereof and V _H comprising the sequence set forth in SEQ ID NO: 160 or CDRs thereof
The BTN2A1 agonist of claim 27(A), which is an antibody comprising

1. A pharmaceutical composition for preventing, treating, slowing progression, preventing recurrence or ameliorating symptoms of cancer or an infectious disease in a subject in need thereof, comprising:
29. A pharmaceutical composition comprising the BTN2A1 agonist of claim 27 or 28.