JP2021019626A5

JP2021019626A5 -

Info

Publication number: JP2021019626A5
Application number: JP2020177784A
Authority: JP
Filing date: 2020-10-23
Publication date: 2021-08-26

Claims

Comprising a first polypeptide chain and a second polypeptide chain, a bispecific polypeptide molecule,
a) The first polypeptide chain is
ai) First variable domain of antibody (VD1),
aii) T first variable domain of a cell receptor (TCR) (VR1), and
aiii) A first linker (LINK1) that links the two domains;
Including
b) The second polypeptide chain is
bi) Second variable domain of TCR (VR2),
bii) Second variable domain of antibody (VD2), and
biii) A second linker (LINK2) that links the two domains;
Including
The first binding site (VD1) (VD2) to which the first variable domain (VD1) and the second variable domain (VD2) bind to specifically bind to the cell surface antigen of human immune effector cells is formed. Form;
The first variable domain (VR1) is either the TCR Vα domain or the TCR Vβ domain, and the second variable domain (VR2) is the other TCR Vα domain that is not selected in the first variable domain (VR1). Or one of the TCR Vβ domains,
The first variable domain (VR1) and the second variable domain (VR2) bind to form a second binding site (VR1) (VR2) that specifically binds to the MHC-related peptide epitope;
The two polypeptide chains are fused to the human IgG hinge domain and / or the human IgG Fc domain or a dimerized portion thereof;
The two polypeptide chains are linked by covalent and / or non-covalent bonds between the hinge domain and / or Fc-domain;
The bispecific polypeptide molecule can simultaneously bind to the cell surface molecule and the MHC-related peptide epitope.
The order of the variable domains in the two polypeptide chains is selected from VD1-VR1 and VR2-VD2 or VD2-VR2 and VR1-VD1 .
(I) The first variable domain of the antibody (VD1) and the second variable domain of the antibody (VD2) contain or contain the variable domain of the α / βTCR-specific antibody BMA031.
The first variable domain of the antibody (VD1) and the second variable domain of the antibody (VD2) include the variable domain of the α / βTCR-specific antibody BMA031 and the first variable domain of the antibody (VD1). ) And the second variable domain of the antibody (VD2) contain, or have an amino acid sequence that is at least 90% identical to the variable domain of the α / βTCR-specific antibody BMA031.
(Ii) The first variable domain of the antibody (VD1) and the second variable domain of the antibody (VD2) contain or contain the variable domain of the humanized T cell mobilization antibody hUCHT1.
The first variable domain of the antibody (VD1) and the second variable domain of the antibody (VD2) include the variable domain of the humanized T cell mobilization antibody hUCHT1 and the first variable domain of the antibody (VD1). ) And the second variable domain of the antibody (VD2) comprises an amino acid sequence having at least 70% identity with the variable domain of the humanized T cell mobilizing antibody hUCHT1.
Bispecific polypeptide molecule.

The bispecific polypeptide molecule of claim 1, wherein the linker sequences LINK1 and / or LINK2 contain at least one sequence motif selected from GGGS, GGGGS, TVLRT, TVSSAS, and TVLSSAS.

The first and second polypeptide chains comprises at least an additional hinge domain and Fc domain or portion thereof derived from human IgG1, IgG2 or IgG4.
The first and second polypeptide chains further comprise at least a hinge domain and Fc domain derived from human IgG1, IgG2 or IgG4 or a portion thereof, wherein the Fc domain is the sequence of SEQ ID NO: 50 (ELLGGP) of human IgG1. Contains at least one effector function silencing mutation in, or
The first and second polypeptide chains further comprise at least a hinge domain and Fc domain derived from human IgG1, IgG2 or IgG4 or a portion thereof, wherein the Fc domain is the sequence of SEQ ID NO: 50 (ELLGGP) of human IgG1. It contains at least one effector function silencing mutation, said effector function silencing mutation generated by one or more mutations in SEQ ID NO: 50 with no residue at E1P, L2V, L3A, or position 4. Be done,
The bispecific polypeptide molecule of claim 1 or 2.

The Fc domain
(I) Containing or containing a CH3 domain comprising at least one mutation that promotes the formation of a heterodimer.
(Ii) Containing a CH3 domain comprising at least one mutation that promotes the formation of a heterodimer, said mutation comprising a knob-in-hole mutation.
Bispecific polypeptide molecule of claim 3.

The Fc domain comprising CH2 and CH3 domains comprising at least two additional cysteine residues, the bispecific polypeptide molecule of claim 3 or 4.

The antibody-derived domains VD1 and VD2 present an engineered disulfide bridge that introduces a covalent bond between VD1 and VD2, where the cysteine is in framework region (FR) 4 when VL and VH. The bispecific polypeptide molecule according to any one of claims 1 to 5 , which is introduced into the framework region 2.

The cell surface molecules are known to induce activation of immune cells, or
The cell surface molecule
(I) CD3, CD4, CD7, CD8, CD10, CD11b, CD11c, CD14, CD16, CD18, CD22, CD25, CD28, CD32a, CD32b, CD33, CD41, CD41b, CD42a, CD42b, CD44, CD45RA, CD49, CD55 , CD56, CD61, CD64, CD68, CD94, CD90, CD117, CD123, CD125, CD134, CD137, CD152, CD163, CD193, CD203c, CD235a, CD278, CD279, CD287, Nkp46, NKG2D, GITR, FcεRI, TCRα / β And TCRγ / δ, HLA-DR or
(Ii) The bispecific polypeptide molecule according to any one of claims 1 to 6 , which is at least one selected from the group consisting of CD3γ, CD3δ, or CD3ε chain.

(I) The bispecific polypeptide molecule comprises a sequence that is at least 70% identical to the sequence set forth in SEQ ID NO: 29, and / or the bispecific polypeptide molecule is the sequence set forth in SEQ ID NO: 32. Containing sequences having at least 90% identity with and / or
(Ii) The bispecific polypeptide molecule comprises a sequence that is at least 90% identical to the sequence set forth in SEQ ID NO: 28, and / or the bispecific polypeptide molecule is the sequence set forth in SEQ ID NO: 31. Containing a sequence having at least 90% identity with
The bispecific polypeptide molecule according to any one of claims 1 to 6.

Said first polypeptide chain, SEQ ID NO: 28, SEQ ID NO: 29, and comprises SEQ ID NO: 30; and the second polypeptide chain, SEQ ID NO: 31, SEQ ID NO: 32, comprising SEQ ID NO: 30 The bispecific polypeptide molecule according to any one of claims 1 to 8.

The Fc region in the first polypeptide chain comprises SEQ ID NO: 26 or SEQ ID NO: 47 (Fc1), and the Fc region in the second polypeptide chain is SEQ ID NO: 27 or SEQ ID NO: 48. (Fc2) comprising a bispecific polypeptide molecule according to any one of claims 3-9.

The first polypeptide chain, SEQ ID NO: 16, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 51, SEQ ID NO: 53, see containing the SEQ ID NO: 55 or SEQ ID NO: 57, and a second polypeptide chain, the sequence No. 17, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56 or SEQ ID NO: 58 including, bispecific polypeptide molecule of claim 1.

The first binding site (VD1) (VD2) that binds to the cell surface antigen of the immune cell is humanized; and / or the second binding site (VR1) (VR2) that binds to the MHC-related peptide epitope. The bispecific polypeptide molecule according to any one of claims 1 to 11 , wherein is affinity maturation and / or stability maturation.

An expression vector comprising the nucleic acid encoding the first polypeptide chain and / or the second polypeptide chain according to any one of claims 1 to 12, or at least one of the nucleic acids.

A host cell comprising the vector according to claim 13 and optionally expressing it.

Together with one or more pharmaceutically acceptable carriers or excipients, the bispecific polypeptide molecule according to any one of claims 1 to 12 nucleic acid or expression vector according to claim 13, Alternatively, a pharmaceutical composition comprising the cell according to claim 14.

Cancer, infectious diseases, and for use in the prevention or treatment of a disease or disorder selected from an immunological disorder, bispecific polypeptide molecule according to any one of claims 1 to 12 The nucleic acid or expression vector of claim 13 , the cell of claim 14 , or the pharmaceutical composition of claim 15.