US2022162341A1PendingUtilityA1
Methods of using monoclonal antibodies targeting epitopes of asph
Est. expiryJun 18, 2038(~11.9 yrs left)· nominal 20-yr term from priority
Inventors:Mark Jon Olsen
G01N 33/575C07K 2317/73C07K 2317/24C07K 2317/92C07K 2317/34C07K 2317/32C07K 16/40C07K 16/2827C07K 2317/565C07K 16/2818
73
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Claims
Abstract
Monoclonal antibodies (MAbs) targeting one or more specific epitopes of aspartyl (asparaginyl) hydroxylase (ASPH), including humanized, bi-specific, and other chimeric MAb variants, and fragments thereof (collectively ASPH epitope-specific MAbs, or simply ASPH MAbs), are disclosed. Methods of production, purification, and use of the ASPH epitope-specific MAbs, and compositions comprising them, as agents in therapeutic and diagnostic applications to interact with target molecules in cell-free samples, cell- and tissue-based assays, animal models, and in a subject are also disclosed. Other aspects of the invention relate to use of the molecules disclosed herein to diagnose, ameliorate, or treat cell proliferation disorders and related diseases.
Claims
exact text as granted — not AI-modifiedWhat is claimed is:
1 . A composition comprising two or more humanized antibodies or fragments or variants thereof, wherein one humanized antibody or fragment or variant thereof, targets at least one epitope of ASPH and another humanized antibody or fragment or variant thereof targets at least one epitope selected from the group consisting of
the T-cell redirector class, comprising an antibody targeting one or more ASPH CDRs and an antibody targeting CD3; the NK-cell redirector class, comprising an antibody targeting one or more ASPH CDRs and an antibody targeting CD16A; the tumor targeting immunomodular class, comprising an antibody targeting one or more ASPH CDRs and an antibody targeting CD40 or 4-1BB; and the dual immunomodular class, comprising an antibody targeting one or more ASPH CDRs and an antibody targeting PD-L1, PD-1, CTLA-4, TGF-β, LAG-3, TIM-3, or OX40.
2 . The composition of claim 1 , wherein said humanized antibody or fragment or variant thereof, which binds to one or more peptide epitopes of human aspartyl (asparaginyl) β-hydroxylase (ASPH) comprises:
a recombinant heavy chain and a recombinant light chain, each heavy and each light chain comprising 3 complementarity-determining regions (CDRs), or fragment or variant thereof, which binds to one or more peptide epitopes of human aspartyl (asparaginyl) β-hydroxylase (ASPH), wherein at least one of said peptide epitopes comprises at least 4 consecutive amino acid residues located within or adjacent to a position in the catalytic domain of ASPH that is within 30 amino acids of the C-terminus of human ASPH, corresponding to the sequence
QDASSFRLIFIVDVWHPELTPQQRRSLPAI
represented by positions
729-758 of SEQ ID NO: 1;
wherein said antibody or fragment or variant thereof contains one or more conservative amino acid substitutions in which the functional activity relating to binding of the antibody or fragment thereof to an epitope of ASPH is retained;
wherein said antibody comprises a recombinant heavy chain comprising
a CDR1 comprising a sequence selected from the group consisting of
NFMC represented by SEQ ID NO: 31,
and
NAMC represented by SEQ ID NO: 32;
a CDR2 comprising a sequence selected from the group consisting of
CIYF
represented by SEQ ID NO: 33,
and
CIDN
represented by SEQ ID NO: 34;
and
a CDR3 comprising a sequence selected from the group consisting of
DGPGSISWKI
represented by SEQ ID NO: 35,
and
NFNI
represented by SEQ ID NO: 36;
wherein said antibody comprises a recombinant light chain comprising
a CDR1 comprising a sequence selected from the group consisting of
SVYSKNR
represented by SEQ ID NO: 37,
and
SVYDNNR
represented by SEQ ID NO: 38;
a CDR2 comprising the sequence
LAS
represented by SEQ ID NO: 39;
and
a CDR3 comprising a sequence selected from the group consisting of
QGTYDSSGWYWA
represented by SEQ ID NO: 40,
and
LGSYSGYIYI
represented by SEQ ID NO: 41.
3 . The composition of claim 2 , wherein said humanized antibody or fragment or variant that targets at least one epitope of ASPH binds to one or more peptides selected from the group consisting of
(a) a peptide comprising 29 amino acids with Cysteine at its amino terminus, plus 28 amino acids corresponding to positions 731-758 at the C-terminal end of human ASPH represented by SEQ ID NO: 1, with the Threonine at relative position 19, corresponding to position 748 of human ASPH, phosphorylated, as
CASSFRLIFIVDVWHPEL-T(PO 3 H 2 )-PQQRRSLPAI
represented by SEQ ID NO: 19;
and
(b) a peptide comprising 29 amino acids with Cysteine at its amino terminus, plus 28 amino acids corresponding to positions 731-758 at the C-terminal end of human ASPH represented by SEQ ID NO: 1, as
CASSFRLIFIVDVWHPELTPQQRRSLPAI,
represented by SEQ ID NO: 20.
4 . The composition of claim 2 , wherein said antibody humanized antibody or fragment or variant that targets at least one epitope of ASPH comprises a recombinant heavy chain and a recombinant light chain, or a fragment thereof, which binds to one or more peptide epitopes of human aspartyl (asparaginyl) β-hydroxylase (ASPH), wherein at least one of said peptide epitopes is located within or adjacent to a position in the catalytic domain of ASPH that is within 30 amino acids of the C-terminus of human ASPH, corresponding to the sequence
QDASSFRLIFIVDVWHPELTPQQRRSLPAI
represented by positions
729-758 of SEQ ID NO: 1;
wherein said antibody binds to an epitope comprising at least 4 consecutive amino acid residues located within 30 amino acids from the C-terminal end of human ASPH represented by SEQ ID NO: 1;
wherein said epitope comprising at least 4 consecutive amino acid residues located within 30 amino acids from the C-terminal end of human ASPH comprises the consecutive amino acid residues selected from the group consisting of
PELT represented by SEQ ID NO: 42,
ELTP represented by SEQ ID NO: 43,
LTPQ represented by SEQ ID NO: 44,
TPQQ represented by SEQ ID NO: 45,
PQQR represented by SEQ ID NO: 46,
QQRR represented by SEQ ID NO: 47,
QRRS represented by SEQ ID NO: 48,
RRSL represented by SEQ ID NO: 49,
RSLP represented by SEQ ID NO: 50,
SLPA represented by SEQ ID NO: 51,
and
LPAI represented by SEQ ID NO: 52.
5 . A composition comprising at least one humanized bispecific antibody, or fragment or variant thereof, wherein said antibody or fragment or variant thereof comprises one or more complementarity determining regions (CDRs) derived from a non-human source targeting one or more peptide epitopes located within or adjacent to the catalytic domain of ASPH, and an antibody targeting other epitopes selected from the group consisting of
the T-cell redirector class, comprising an antibody targeting one or more ASPH CDRs and an antibody targeting CD3; the NK-cell redirector class, comprising an antibody targeting one or more ASPH CDRs and an antibody targeting CD16A; the tumor targeting immunomodular class, comprising an antibody targeting one or more ASPH CDRs and an antibody targeting CD40 or 4-1BB; and the dual immunomodular class, comprising an antibody targeting one or more ASPH CDRs and an antibody targeting PD-L1, PD-1, CTLA-4, TGF-β, LAG-3, TIM-3, or OX40.
6 . The composition of claim 5 , wherein said humanized antibody or fragment or variant thereof, which binds to one or more peptide epitopes of human aspartyl (asparaginyl) β-hydroxylase (ASPH) comprises:
a recombinant heavy chain and a recombinant light chain, each heavy and each light chain comprising 3 complementarity-determining regions (CDRs), or fragment or variant thereof, which binds to one or more peptide epitopes of human aspartyl (asparaginyl) β-hydroxylase (ASPH), wherein at least one of said peptide epitopes comprises at least 4 consecutive amino acid residues located within or adjacent to a position in the catalytic domain of ASPH that is within 30 amino acids of the C-terminus of human ASPH, corresponding to the sequence
QDASSFRLIFIVDVWHPELTPQQRRSLPAI
represented by positions
729-758 of SEQ ID NO: 1;
wherein said antibody or fragment or variant thereof contains one or more conservative amino acid substitutions in which the functional activity relating to binding of the antibody or fragment thereof to an epitope of ASPH is retained;
wherein said antibody comprises a recombinant heavy chain comprising
a CDR1 comprising a sequence selected from the group consisting of
NFMC
represented by SEQ ID NO: 31,
and
NAMC
represented by SEQ ID NO: 32;
a CDR2 comprising a sequence selected from the group consisting of
CIYF
represented by SEQ ID NO: 33,
and
CIDN
represented by SEQ ID NO: 34;
and
a CDR3 comprising a sequence selected from the group consisting of
DGPGSISWKI
represented by SEQ ID NO: 35,
and
NFNI
represented by SEQ ID NO: 36;
wherein said antibody comprises a recombinant light chain comprising a CDR1 comprising a sequence selected from the group consisting of
SVYSKNR
represented by SEQ ID NO: 37,
and
SVYDNNR
represented by SEQ ID NO: 38;
a CDR2 comprising the sequence
LAS represented by SEQ ID NO: 39;
and
a CDR3 comprising a sequence selected from the group consisting of
QGTYDSSGWYWA
represented by SEQ ID NO: 40,
and
LGSYSGYIYI
represented by SEQ ID NO: 41.
7 . The composition of claim 6 , wherein said humanized antibody or fragment or variant that targets at least one epitope of ASPH binds to one or more peptides selected from the group consisting of
(a) a peptide comprising 29 amino acids with Cysteine at its amino terminus, plus 28 amino acids corresponding to positions 731-758 at the C-terminal end of human ASPH represented by SEQ ID NO: 1, with the Threonine at relative position 19, corresponding to position 748 of human ASPH, phosphorylated, as
CASSFRLIFIVDVWHPEL-T(PO 3 H 2 )-PQQRRSLPAI
represented by SEQ ID NO: 19;
and
(b) a peptide comprising 29 amino acids with Cysteine at its amino terminus, plus 28 amino acids corresponding to positions 731-758 at the C-terminal end of human ASPH represented by SEQ ID NO: 1, as
CASSFRLIFIVDVWHPELTPQQRRSLPAI,
represented by SEQ ID NO: 20.
8 . The composition of claim 6 , wherein said antibody humanized antibody or fragment or variant that targets at least one epitope of ASPH comprises a recombinant heavy chain and a recombinant light chain, or a fragment thereof, which binds to one or more peptide epitopes of human aspartyl (asparaginyl) hydroxylase (ASPH), wherein at least one of said peptide epitopes is located within or adjacent to a position in the catalytic domain of ASPH that is within 30 amino acids of the C-terminus of human ASPH, corresponding to the sequence
QDASSFRLIFIVDVWHPELTPQQRRSLPAI
represented by positions
729-758 of SEQ ID NO: 1;
wherein said antibody binds to an epitope comprising at least 4 consecutive amino acid residues located within 30 amino acids from the C-terminal end of human ASPH represented by SEQ ID NO: 1;
wherein said epitope comprising at least 4 consecutive amino acid residues located within 30 amino acids from the C-terminal end of human ASPH comprises the consecutive amino acid residues selected from the group consisting of
PELT represented by SEQ ID NO: 42,
ELTP represented by SEQ ID NO: 43,
LTPQ represented by SEQ ID NO: 44,
TPQQ represented by SEQ ID NO: 45,
PQQR represented by SEQ ID NO: 46,
QQRR represented by SEQ ID NO: 47,
QRRS represented by SEQ ID NO: 48,
RRSL represented by SEQ ID NO: 49,
RSLP represented by SEQ ID NO: 50,
SLPA represented by SEQ ID NO: 51,
and
LPAI represented by SEQ ID NO: 52.Cited by (0)
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