CA2510180C - Humanized antibody (h14.18) of the mouse 14.18 antibody binding to gd2 and its fusion with il-2 - Google Patents
Humanized antibody (h14.18) of the mouse 14.18 antibody binding to gd2 and its fusion with il-2 Download PDFInfo
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- CA2510180C CA2510180C CA2510180A CA2510180A CA2510180C CA 2510180 C CA2510180 C CA 2510180C CA 2510180 A CA2510180 A CA 2510180A CA 2510180 A CA2510180 A CA 2510180A CA 2510180 C CA2510180 C CA 2510180C
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
- C07K16/3076—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells against structure-related tumour-associated moieties
- C07K16/3084—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells against structure-related tumour-associated moieties against tumour-associated gangliosides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/46—Hybrid immunoglobulins
- C07K16/461—Igs containing Ig-regions, -domains or -residues form different species
- C07K16/467—Igs with modifications in the FR-residues only
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
- C07K2317/732—Antibody-dependent cellular cytotoxicity [ADCC]
Abstract
The invention provides humanized antibody H14.18 binding the human cell surface glycosphingolipid GD2. The antibody comprises modified variable regions, more specially, modified framework regions, which reduce their immunogenitity when administered to a human. The antibody may be coupled to the therapeutic agent such as IL-2 and used in the treatment of cancer.
Description
HUMANIZED ANTIBODY (H14.18) OF THE MOUSE 14.18 ANTIBODY BINDING TO GD2 AND ITS
This invention relates generally to modified antibodies. More particularly, the invention relates to modified antibodies with reduced immunogenicity that specifically bind the human cell surface glycosphingolipid GD2, and their use as therapeutic agents.
BACKGROUND OF THE INVENTION
There has been significant progress in the development of antibody-based therapies over the years. For example, investigators have identified not only a variety of cancer-specific markers but also a variety of antibodies that specifically bind to those markers. Antibodies can be used to deliver certain molecules, for example, a toxin or an immune stimulatory moiety, for example, a cytokine, to a cancer cell expressing the marker so as to selectively kill the cancer cell.
The 14.18 antibody is a mouse-derived monoclonal antibody directed against the cell surface glycosphingolipid GD2. GD2 is a disialoganglioside that is normally only expressed at a significant level on the outer surface membranes of neuronal cells, where its exposure to the immune system is limited by the blood brain barrier.
Many tumor cells, in contrast, have abnormal levels of glycosphingolipid cell surface expression. For example, GD2 is expressed on the surfaces of a wide range of tumor cells including neuroblastomas, medulloblastomas, astrocytomas, melanomas, small-cell lung cancer, osteosarcomas and other soft tissue sarcomas. Thus, GD2 is a convenient tumor-specific marker for targeting immune-stimulatory protein domains to tumor cells for the purpose of raising an effective immune response against the tumor cells to destroy them. While the 14.18 mouse antibody (ml4.18 antibody) may assist the targeting of these protein domains to tumor cells, its mouse-derived amino acid sequences can impair the desired therapeutic effect.
When administered to a patient, antibodies can have an associated immunogenicity in the host mammal. This is more likely to occur when the antibodies are not autologous. Consequently, the effectiveness of antibody-based therapies often is limited by an immunogenic response directed against the therapeutic antibody.
This immunogenic response typically is increased when the antibody is derived in whole- or in part from a mammal different than the host mammal, e.g., when the antibody is derived from a mouse and the recipient is a human.
This invention relates generally to modified antibodies. More particularly, the invention relates to modified antibodies with reduced immunogenicity that specifically bind the human cell surface glycosphingolipid GD2, and their use as therapeutic agents.
BACKGROUND OF THE INVENTION
There has been significant progress in the development of antibody-based therapies over the years. For example, investigators have identified not only a variety of cancer-specific markers but also a variety of antibodies that specifically bind to those markers. Antibodies can be used to deliver certain molecules, for example, a toxin or an immune stimulatory moiety, for example, a cytokine, to a cancer cell expressing the marker so as to selectively kill the cancer cell.
The 14.18 antibody is a mouse-derived monoclonal antibody directed against the cell surface glycosphingolipid GD2. GD2 is a disialoganglioside that is normally only expressed at a significant level on the outer surface membranes of neuronal cells, where its exposure to the immune system is limited by the blood brain barrier.
Many tumor cells, in contrast, have abnormal levels of glycosphingolipid cell surface expression. For example, GD2 is expressed on the surfaces of a wide range of tumor cells including neuroblastomas, medulloblastomas, astrocytomas, melanomas, small-cell lung cancer, osteosarcomas and other soft tissue sarcomas. Thus, GD2 is a convenient tumor-specific marker for targeting immune-stimulatory protein domains to tumor cells for the purpose of raising an effective immune response against the tumor cells to destroy them. While the 14.18 mouse antibody (ml4.18 antibody) may assist the targeting of these protein domains to tumor cells, its mouse-derived amino acid sequences can impair the desired therapeutic effect.
When administered to a patient, antibodies can have an associated immunogenicity in the host mammal. This is more likely to occur when the antibodies are not autologous. Consequently, the effectiveness of antibody-based therapies often is limited by an immunogenic response directed against the therapeutic antibody.
This immunogenic response typically is increased when the antibody is derived in whole- or in part from a mammal different than the host mammal, e.g., when the antibody is derived from a mouse and the recipient is a human.
For clinical use in humans, it may be helpful to modify mouse-derived antibodies to more closely resemble human antibodies, so as to reduce or minimize the immunogenicity of the mouse-derived antibody. The immunogenicity of the mouse-derived antibody can be reduced by the creation of a chimeric antibody in which the constant regions of a human antibody are fused to mouse variable domains.
However, the remaining mouse variable domains are generally still immunogenic in humans, and can thus impair the efficacy of an antibody-based therapy.
Some approaches to reducing immunogenicity, such as "veneering" and "humanization" involve the introduction of many amino acid substitutions and may 1o disrupt the binding of an antibody to an antigen. The m14.18 antibody binds to GD2 with moderate affinity. Therefore, mutations that significantly lower the affinity of m14.18 for GD2 are expected to make it less effective for therapeutic purposes in humans.
Accordingly, there is a need in the art for therapeutic antibodies that can effectively target GD2 and have reduced immunogenicity when administered to a human.
SUMMARY OF THE INVENTION
Generally, the present invention provides a modified form of the ml4.18 antibody that is less immunogenic in humans, but still maintains the binding affinity of m14.18 for human GD2.
More particularly, the invention provides a humanized form of the ml4.18 antibody (hu14.18 antibody) in which several mouse-specific amino acids in one or more of the framework regions have been substituted with different amino acids to reduce their immunogenicity in humans. The invention also provides fusions of the hul4.18 antibody to one or more non-immunoglobulin moieties for enhancing the effects of targeted immune therapy.
In one aspect, the present invention provides an antibody variable region including the amino acid sequence set forth in SEQ ID NO: 1, which defines an immunoglobulin light chain variable region (VL region). In another aspect, the invention relates to an antibody variable region including the amino acid sequence set forth in SEQ ID
NO: 2, which defines an immunoglobulin heavy chain variable region (VH region). In one embodiment, the invention provides an antibody variable region in which the amino acid sequence of SEQ ID NO: 1 is linked to the amino acid sequence set forth in SEQ
ID NO:
However, the remaining mouse variable domains are generally still immunogenic in humans, and can thus impair the efficacy of an antibody-based therapy.
Some approaches to reducing immunogenicity, such as "veneering" and "humanization" involve the introduction of many amino acid substitutions and may 1o disrupt the binding of an antibody to an antigen. The m14.18 antibody binds to GD2 with moderate affinity. Therefore, mutations that significantly lower the affinity of m14.18 for GD2 are expected to make it less effective for therapeutic purposes in humans.
Accordingly, there is a need in the art for therapeutic antibodies that can effectively target GD2 and have reduced immunogenicity when administered to a human.
SUMMARY OF THE INVENTION
Generally, the present invention provides a modified form of the ml4.18 antibody that is less immunogenic in humans, but still maintains the binding affinity of m14.18 for human GD2.
More particularly, the invention provides a humanized form of the ml4.18 antibody (hu14.18 antibody) in which several mouse-specific amino acids in one or more of the framework regions have been substituted with different amino acids to reduce their immunogenicity in humans. The invention also provides fusions of the hul4.18 antibody to one or more non-immunoglobulin moieties for enhancing the effects of targeted immune therapy.
In one aspect, the present invention provides an antibody variable region including the amino acid sequence set forth in SEQ ID NO: 1, which defines an immunoglobulin light chain variable region (VL region). In another aspect, the invention relates to an antibody variable region including the amino acid sequence set forth in SEQ ID
NO: 2, which defines an immunoglobulin heavy chain variable region (VH region). In one embodiment, the invention provides an antibody variable region in which the amino acid sequence of SEQ ID NO: 1 is linked to the amino acid sequence set forth in SEQ
ID NO:
2. The amino acid sequences can be linked, such as by a disulfide bond or a peptide bond.
In another aspect, the invention relates to an antibody variable region that specifically binds to GD2 and includes at least amino acids 1-23 of SEQ ID NO:
1, amino acids 1.-25 of SEQ ID NO: 2, or amino acids 67-98 of SEQ ID NO: 2. These sequences define framework regions in the immunoglobulin variable regions of the hu14.18 antibody. Framework regions are described in greater detail below.
One aspect of the invention relates to a method for targeting a cell with GD2 on its surface and includes administering an antibody variable region of the present invention to a patient. In one embodiment, the targeted cell is a tumor cell. Further aspects of the invention include a nucleic acid encoding the antibody variable region or a cell that includes this nucleic acid, either of which can be administered to a patient or used for in vitro protein production.
The invention also provides a polypeptide that includes an antibody variable region of the invention and an Fc portion comprising at least a CH2 domain, nucleic acids encoding the polypeptide, cells including the nucleic acids, and methods for targeting a cell with GD2 on its surface by administering the polypeptide, nucleic acid, or cell to a patient. In some embodiments of the invention, the Fc portion is derived from IgGi.
The antibody variable region can be linked, with or without an intervening Fc portion, to a non-immunoglobulin moiety. Specifically, the non-immunoglobulin moiety can be a cytokine, such as an interleukin, a hematopoietic factor, a lympholdne, an interferon, or a chemokine. The interleukin can be, for example, interleukin-2 or interleuk-in-12. The hematopoietic factor and lymphokine can be, for example, granulocyte-macrophage colony stimulating factor (GM-CSF) and a lymphotoxin, respectively. The interferon can be, for example, interferon-a, interferon-fl, or interferon-y. In some embodiments of the invention, the fusion protein includes a second non-immunologlobulin moiety, such as a second cytokine. In a particular embodiment, the fusion protein includes the antibody variable region, IL-2, and IL-12.
3a In one aspect, the invention provides a humanized antibody-1L2 fusion protein designated as hu14.18-1L2 that specifically binds GD2 and stimulates immune function comprising the light chain of SEQ ID NO. 5 and the heavy chain of SEQ
ID
NO. 6.
In another aspect, the invention provides a vector comprising the nucleotide sequence of SEQ ID NO. 4 containing the nucleic acid sequences that code for the fusion protein as described above.
In another aspect, the invention provides pharmaceutical composition comprising the fusion protein as described above and a pharmaceutical carrier or excipient.
In another aspect, the invention provides use of the fusion protein as described above for the manufacture of a medicament for stabilizing disease progression in GD2 positive cancer patients.
In another aspect, the invention provides use of the fusion protein as described above for the manufacture of a medicament for increasing ADCC and NK-lysis activity in GD2 positive cancer patients.
In another aspect, the invention provides a pharmaceutical composition as described above, for use in stabilizing disease progression in GD2 positive cancer patients.
In another aspect, the invention provides a pharmaceutical composition as described above, for use in increasing ADCC and NK-lysis activity in GD2 positive cancer patients.
It is to be understood that the features of the various embodiments described herein are not mutually exclusive and can exist in various combinations and permutations.
3b Description of the Drawings Figure 1A shows the amino acid sequence of an immunoglobulin light chain variable region in accordance with the invention.
In another aspect, the invention relates to an antibody variable region that specifically binds to GD2 and includes at least amino acids 1-23 of SEQ ID NO:
1, amino acids 1.-25 of SEQ ID NO: 2, or amino acids 67-98 of SEQ ID NO: 2. These sequences define framework regions in the immunoglobulin variable regions of the hu14.18 antibody. Framework regions are described in greater detail below.
One aspect of the invention relates to a method for targeting a cell with GD2 on its surface and includes administering an antibody variable region of the present invention to a patient. In one embodiment, the targeted cell is a tumor cell. Further aspects of the invention include a nucleic acid encoding the antibody variable region or a cell that includes this nucleic acid, either of which can be administered to a patient or used for in vitro protein production.
The invention also provides a polypeptide that includes an antibody variable region of the invention and an Fc portion comprising at least a CH2 domain, nucleic acids encoding the polypeptide, cells including the nucleic acids, and methods for targeting a cell with GD2 on its surface by administering the polypeptide, nucleic acid, or cell to a patient. In some embodiments of the invention, the Fc portion is derived from IgGi.
The antibody variable region can be linked, with or without an intervening Fc portion, to a non-immunoglobulin moiety. Specifically, the non-immunoglobulin moiety can be a cytokine, such as an interleukin, a hematopoietic factor, a lympholdne, an interferon, or a chemokine. The interleukin can be, for example, interleukin-2 or interleuk-in-12. The hematopoietic factor and lymphokine can be, for example, granulocyte-macrophage colony stimulating factor (GM-CSF) and a lymphotoxin, respectively. The interferon can be, for example, interferon-a, interferon-fl, or interferon-y. In some embodiments of the invention, the fusion protein includes a second non-immunologlobulin moiety, such as a second cytokine. In a particular embodiment, the fusion protein includes the antibody variable region, IL-2, and IL-12.
3a In one aspect, the invention provides a humanized antibody-1L2 fusion protein designated as hu14.18-1L2 that specifically binds GD2 and stimulates immune function comprising the light chain of SEQ ID NO. 5 and the heavy chain of SEQ
ID
NO. 6.
In another aspect, the invention provides a vector comprising the nucleotide sequence of SEQ ID NO. 4 containing the nucleic acid sequences that code for the fusion protein as described above.
In another aspect, the invention provides pharmaceutical composition comprising the fusion protein as described above and a pharmaceutical carrier or excipient.
In another aspect, the invention provides use of the fusion protein as described above for the manufacture of a medicament for stabilizing disease progression in GD2 positive cancer patients.
In another aspect, the invention provides use of the fusion protein as described above for the manufacture of a medicament for increasing ADCC and NK-lysis activity in GD2 positive cancer patients.
In another aspect, the invention provides a pharmaceutical composition as described above, for use in stabilizing disease progression in GD2 positive cancer patients.
In another aspect, the invention provides a pharmaceutical composition as described above, for use in increasing ADCC and NK-lysis activity in GD2 positive cancer patients.
It is to be understood that the features of the various embodiments described herein are not mutually exclusive and can exist in various combinations and permutations.
3b Description of the Drawings Figure 1A shows the amino acid sequence of an immunoglobulin light chain variable region in accordance with the invention.
Figure 1B shows the amino acid sequence of an immunoglobulin heavy chain variable region in accordance with the invention.
Figure 2A-D show the nucleotide sequence of an expression vector, including the nucleic acid constructs encoding an immunoglobulin light chain and an immunoglobulin heavy chain-IL-2 fusion protein in accordance with the invention.
Figure 3A shows the amino acid sequence of an immunoglobulin light chain in accordance with the invention.
Figure 3B shows the amino acid sequence of an immunoglobulin heavy chain in accordance with the invention.
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides a modified form of the m14.18 antibody that is less immunogenic in humans, but is still able to specifically bind human GD2.
The reduced immunogenicity is provided by one or more altered amino acid sequences in the immunoglobulin variable domains. The antibody is useful for treating GD2-positive tumors, particularly when fused to a cytokine or other immune modulator.
As used herein, the terms "antibody" and "immunoglobulin" are understood to mean (i) an intact antibody (for example, a monoclonal antibody or polyclonal antibody), (ii) antigen binding portions thereof, including, for example, an Fab fragment, an Fab' fragment, an (Fab')2 fragment, an Fv fragment, a single chain antibody binding site, an sFv, (iii) bi-specific antibodies and antigen binding portions thereof, and (iv) multi-specific antibodies and antigen binding portions thereof.
As used herein, the terms "bind specifically," "specifically bind" and "specific binding" are understood to mean that the antibody has a binding affinity for a particular antigen of at least about 106 M-1, more preferably, at least about 107 M"1, more preferably at least about 108 M"1, and most preferably at least about 1010 M-1.
As used herein, the terms "Framework Regions" and "FRs" are understood to mean the regions of an immunoglobulin variable region adjacent to the Complementarity-Determining Regions (CDRs). CDRs are the portions of an immunoglobulin variable region that interact primarily with an antigen. As shown in FIG. 1, the VH and VL regions both contain four FRs and are located within the boxed portions of the amino acid sequences.
In particular, with reference to the amino acid sequence shown in FIG. 1A (SEQ
ID NO: 1), the light chain FRs are defined by the amino acid sequences from Aspl to -Cys23 (huVLFR1), from His39 to His54 (huVLFR2), from G1y62 to Cys93 (huVLFR3), and from Phe104 to Lysl 13 (huVLFR4). With reference to the amino acid sequence shown in FIG. 1B (SEQ ID NO: 2), the heavy chain FRs are defined by the amino acid sequences from Glul to Ser25 (huVHFR1), from Trp36 to G1y49 (huVHFR2), from Arg67 5 to Ser98 (huVHFR3), and from Trp103 to Serl 13 (huVHFR4).
Protein sequences of the invention The present invention features antibodies that bind, preferably specifically, to the human cell surface glycosphingolipid GD2 and have modified regions derived from the m14.18 antibody. The VH or VL amino acid sequences (or both) are modified or humanized to reduce their immunogenicity when administered to a human. In accordance with the invention, the m14.18 antibody can be humanized, for example, by using deimmunization methods in which potential T cell epitopes are eliminated or weakened by introduction of mutations that reduce binding of a peptide epitope to an MHC Class II
molecule (see, for example, W098/52976 and W000/34317). Alternatively, non-human T cell epitopes are mutated so that they correspond to human self epitopes that are present in human antibodies (see, for example, U.S. Patent No. 5,712,120). The present invention provides GD2 antibodies having VL and VH regions that include at least one humanized FR sequence, thereby reducing immunogenicity when administered to a human.
I. Heavy and Light Chains Variable Regions As mentioned above, the hul4.18 includes humanized variable regions derived from the m14.18 antibody that maintain specific binding of human GD2 antigen.
In some embodiments of the invention, the VL region of the hul4.18 antibody includes the following polypeptide:
D-V-V-M-T-Q-T-P-L-S-L-P-V-T-P-G-E-P-A-S-I-S-C-R-S-S-Q-S-L-V-H-R-N-G-N-T-Y-L-H-W-Y-L-Q-K-P-G-Q-S-P-K-L-L-I-H-K-V-S-N-R-F-S-G-V-P-D-R-F-S-G-S-G-S-G-T-D-F-T-L-K-I-S-R-V-E-A-E-D-L-G-V-Y-F-C-S-Q-S-T-H-V-P-P-L-T-F-G-A-G-T-K-L-E-L-K (SEQ IDNO: 1).
In particular embodiments, the hul4.18 antibody includes a light chain FR1 that is defined by residues 1 to 23 of SEQ ID NO: 1, namely, D-V-V-M-T-Q-T-P-L-S-L-P-V-T-P-G-E-P-A-S-I-S-C (hUVLFRl).
In other embodiments of the invention, the VH region of the hul4.18 antibody includes the following polypeptide:
E-V-Q-L-V-Q-S-G-A-E-V-E-K-P-G-A-S-V-K-I-S-C-K-A-S-G-S-S-F-T-G-Y-N-M-N-W-V-R-Q-N-I-G-K-S-L-E-W-I-G-A-I-D-P-Y-Y-G-G-T-S-Y-N-Q-K-F-K-G-R-A-T-L-T-V-D-K-S-T-S-T-A-Y-M-H-L-K-S-L-R-S-E-D-T-A-V-Y-Y-C-V-S-G-M-E-Y-W-G-Q-G-T-S-V-T-V-S -S (SEQ ID NO: 2).
In particular embodiments, the hu14.18 antibody includes a heavy chain FRI
that is defined by residues 1 to 25 of SEQ ID NO: 2, namely E-V-Q-L-V-Q-S-G-A-E-V-E-K-P-G-A-S-V-K-I-S-C-K-A-S (huVHFR1).
In further embodiments of the invention, the hu14.18 antibody includes a heavy chain FR3 that is represented by residues 67 to 98 of SEQ ID NO: 2, namely R-A-T-L-T-V-D-K-S-T-S-T-A-Y-M-H-L-K-S-L-R-S-E-D-T-A-V-Y-Y-C-V-S (huVHFR3).
Various combinations of the foregoing embodiments are also within the scope of the present invention. For example, the hu14.18 antibody may include the VL
sequence set forth in SEQ ID NO: 1 and the VH sequence set forth in SEQ ID NO: 2. The VL and VH regions can be linked by a disulfide bond or a peptide bond, depending on how their nucleic acid sequences are constructed. In general, V regions are linked by a disulfide bond when their sequences are encoded on separate DNA constructs. In contrast, the V
regions are typically linked by a peptide bond when their sequences are encoded on a single-chain DNA construct.
The present invention also contemplates an antibody that specifically binds and includes at least a portion of the humanized V regions. For example, the hu14.18 antibody can include a VL region as defined by SEQ ID NO:1 and a VH region having at least one humanized FR, such as huVHFRl or huVHFR2. Alternatively, the antibody of the present invention can include a VH region as defined by SEQ ID NO: 2 and a VL
region having at least one humanized FR, such as huVLFR1. The hu14. i 8 antibody can also include a VH region having at least one humanized FR and/or a VL region having at least one humanized FR.
In certain embodiments of the invention, the light chain variable region and the heavy chain variable region can be coupled, respectively, to a light chain constant region and a heavy chain constant region of an immunoglobulin. The immunoglobulin light chains have constant regions that are designated as either kappa or lambda chains. In a particular embodiment of the invention, the light chain constant region is a kappa chain.
3o The heavy chain constant regions, and various modification and combinations thereof are discussed below in detail.
Figure 2A-D show the nucleotide sequence of an expression vector, including the nucleic acid constructs encoding an immunoglobulin light chain and an immunoglobulin heavy chain-IL-2 fusion protein in accordance with the invention.
Figure 3A shows the amino acid sequence of an immunoglobulin light chain in accordance with the invention.
Figure 3B shows the amino acid sequence of an immunoglobulin heavy chain in accordance with the invention.
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides a modified form of the m14.18 antibody that is less immunogenic in humans, but is still able to specifically bind human GD2.
The reduced immunogenicity is provided by one or more altered amino acid sequences in the immunoglobulin variable domains. The antibody is useful for treating GD2-positive tumors, particularly when fused to a cytokine or other immune modulator.
As used herein, the terms "antibody" and "immunoglobulin" are understood to mean (i) an intact antibody (for example, a monoclonal antibody or polyclonal antibody), (ii) antigen binding portions thereof, including, for example, an Fab fragment, an Fab' fragment, an (Fab')2 fragment, an Fv fragment, a single chain antibody binding site, an sFv, (iii) bi-specific antibodies and antigen binding portions thereof, and (iv) multi-specific antibodies and antigen binding portions thereof.
As used herein, the terms "bind specifically," "specifically bind" and "specific binding" are understood to mean that the antibody has a binding affinity for a particular antigen of at least about 106 M-1, more preferably, at least about 107 M"1, more preferably at least about 108 M"1, and most preferably at least about 1010 M-1.
As used herein, the terms "Framework Regions" and "FRs" are understood to mean the regions of an immunoglobulin variable region adjacent to the Complementarity-Determining Regions (CDRs). CDRs are the portions of an immunoglobulin variable region that interact primarily with an antigen. As shown in FIG. 1, the VH and VL regions both contain four FRs and are located within the boxed portions of the amino acid sequences.
In particular, with reference to the amino acid sequence shown in FIG. 1A (SEQ
ID NO: 1), the light chain FRs are defined by the amino acid sequences from Aspl to -Cys23 (huVLFR1), from His39 to His54 (huVLFR2), from G1y62 to Cys93 (huVLFR3), and from Phe104 to Lysl 13 (huVLFR4). With reference to the amino acid sequence shown in FIG. 1B (SEQ ID NO: 2), the heavy chain FRs are defined by the amino acid sequences from Glul to Ser25 (huVHFR1), from Trp36 to G1y49 (huVHFR2), from Arg67 5 to Ser98 (huVHFR3), and from Trp103 to Serl 13 (huVHFR4).
Protein sequences of the invention The present invention features antibodies that bind, preferably specifically, to the human cell surface glycosphingolipid GD2 and have modified regions derived from the m14.18 antibody. The VH or VL amino acid sequences (or both) are modified or humanized to reduce their immunogenicity when administered to a human. In accordance with the invention, the m14.18 antibody can be humanized, for example, by using deimmunization methods in which potential T cell epitopes are eliminated or weakened by introduction of mutations that reduce binding of a peptide epitope to an MHC Class II
molecule (see, for example, W098/52976 and W000/34317). Alternatively, non-human T cell epitopes are mutated so that they correspond to human self epitopes that are present in human antibodies (see, for example, U.S. Patent No. 5,712,120). The present invention provides GD2 antibodies having VL and VH regions that include at least one humanized FR sequence, thereby reducing immunogenicity when administered to a human.
I. Heavy and Light Chains Variable Regions As mentioned above, the hul4.18 includes humanized variable regions derived from the m14.18 antibody that maintain specific binding of human GD2 antigen.
In some embodiments of the invention, the VL region of the hul4.18 antibody includes the following polypeptide:
D-V-V-M-T-Q-T-P-L-S-L-P-V-T-P-G-E-P-A-S-I-S-C-R-S-S-Q-S-L-V-H-R-N-G-N-T-Y-L-H-W-Y-L-Q-K-P-G-Q-S-P-K-L-L-I-H-K-V-S-N-R-F-S-G-V-P-D-R-F-S-G-S-G-S-G-T-D-F-T-L-K-I-S-R-V-E-A-E-D-L-G-V-Y-F-C-S-Q-S-T-H-V-P-P-L-T-F-G-A-G-T-K-L-E-L-K (SEQ IDNO: 1).
In particular embodiments, the hul4.18 antibody includes a light chain FR1 that is defined by residues 1 to 23 of SEQ ID NO: 1, namely, D-V-V-M-T-Q-T-P-L-S-L-P-V-T-P-G-E-P-A-S-I-S-C (hUVLFRl).
In other embodiments of the invention, the VH region of the hul4.18 antibody includes the following polypeptide:
E-V-Q-L-V-Q-S-G-A-E-V-E-K-P-G-A-S-V-K-I-S-C-K-A-S-G-S-S-F-T-G-Y-N-M-N-W-V-R-Q-N-I-G-K-S-L-E-W-I-G-A-I-D-P-Y-Y-G-G-T-S-Y-N-Q-K-F-K-G-R-A-T-L-T-V-D-K-S-T-S-T-A-Y-M-H-L-K-S-L-R-S-E-D-T-A-V-Y-Y-C-V-S-G-M-E-Y-W-G-Q-G-T-S-V-T-V-S -S (SEQ ID NO: 2).
In particular embodiments, the hu14.18 antibody includes a heavy chain FRI
that is defined by residues 1 to 25 of SEQ ID NO: 2, namely E-V-Q-L-V-Q-S-G-A-E-V-E-K-P-G-A-S-V-K-I-S-C-K-A-S (huVHFR1).
In further embodiments of the invention, the hu14.18 antibody includes a heavy chain FR3 that is represented by residues 67 to 98 of SEQ ID NO: 2, namely R-A-T-L-T-V-D-K-S-T-S-T-A-Y-M-H-L-K-S-L-R-S-E-D-T-A-V-Y-Y-C-V-S (huVHFR3).
Various combinations of the foregoing embodiments are also within the scope of the present invention. For example, the hu14.18 antibody may include the VL
sequence set forth in SEQ ID NO: 1 and the VH sequence set forth in SEQ ID NO: 2. The VL and VH regions can be linked by a disulfide bond or a peptide bond, depending on how their nucleic acid sequences are constructed. In general, V regions are linked by a disulfide bond when their sequences are encoded on separate DNA constructs. In contrast, the V
regions are typically linked by a peptide bond when their sequences are encoded on a single-chain DNA construct.
The present invention also contemplates an antibody that specifically binds and includes at least a portion of the humanized V regions. For example, the hu14.18 antibody can include a VL region as defined by SEQ ID NO:1 and a VH region having at least one humanized FR, such as huVHFRl or huVHFR2. Alternatively, the antibody of the present invention can include a VH region as defined by SEQ ID NO: 2 and a VL
region having at least one humanized FR, such as huVLFR1. The hu14. i 8 antibody can also include a VH region having at least one humanized FR and/or a VL region having at least one humanized FR.
In certain embodiments of the invention, the light chain variable region and the heavy chain variable region can be coupled, respectively, to a light chain constant region and a heavy chain constant region of an immunoglobulin. The immunoglobulin light chains have constant regions that are designated as either kappa or lambda chains. In a particular embodiment of the invention, the light chain constant region is a kappa chain.
3o The heavy chain constant regions, and various modification and combinations thereof are discussed below in detail.
7 =
Ii. Fc portion The antibody variable domains of the present invention are optionally fused to an Fc portion. As used herein, the Fe portion encompasses domains derived from the heavy chain constant region of an immunoglobulin, preferably a human immunoglobulin, including a fragment, analog, variant, mutant or derivative of the constant region. The constant region of an immunoglobulin heavy chain is defined as a naturally-occurring or synthetically produced polypeptide homologous to at least a portion of the C-terminal region of the heavy chain, including the CH1, hinge, CH2, CH3, and, for some heavy chain classes, CH4 domains. The "hinge" region joins the CH1 domain to the CH2-region of an Fc portion. The constant region of the heavy chains of all mammalian immunoglobulins exhibit extensive amino acid sequence similarity. DNA
sequences for these immunoglobulin regions are well known in the art. (See, e.g., Gillies et al. (1989) J.
Immunol. Meth. 125:191).
In the present invention, the Fc portion typically includes at least a CH2 domain.
For example, the Fc portion can include the entire immunoglobulin heavy chain constant region (CHl-hinge-CH2-CH3). Alternatively, the Fc portion can include all or a portion of the hinge region, the CH2 domain and the CH3 domain.
The constant region of an immunoglobulin is responsible for many important antibody effector functions, including Fc receptor (FcR) binding and complement fixation. There are five major classes of the heavy chain constant region, classified as IgA, IgG, IgD, IgE, and IgM, each with characteristic effector functions designated by isotype.
IgG, for example, is separated into four y isotypes: yl, y2, y3, and y4, also known as IgGl, IgG2, IgG3, and IgG4, respectively. IgG molecules can interact with multiple classes of cellular receptors including three classes of Fcy receptors (FcyR) specific for the IgG class of antibody, namely FcyRI, FcyRII, and FcyRIII. The sequences important for the binding of IgG to the FcyR receptors have been reported to be in the CH2 and CH3 domains.
The serum half-life of an antibody is influenced by the ability of that antibody to bind to an Fc receptor (EcR). Similarly, the serum half-life of immunoglobulin fusion proteins is also influenced by the inability to bind to such receptors (Gillies et al., Cancer Research (1999) 59:2159-66). The CH2 and CH3 domains of IgG2 and IgG4 have undetectable or reduced binding affinity to Fe receptors compared to those of IgGl.
Accordingly, the serum half-life of the featured antibody can be increased by using the CH2 and/or CH3 domain from IgG2 or IgG4 isotypes. Alternatively, the antibody can include a CH2 and/or CH3 domain from IgGI or IgG3 with modification in one or more amino acids in these domains to reduce the binding affinity for Fc receptors (see, e.g., U.S. patent application 09/256,156, published as U.S. patent application publication 2003-0105294-Al).
The hinge region of the Fc portion normally adjoins the C-terminus of the CHI
domain of the heavy chain constant region. When included in the proteins of the present invention, the hinge is homologous to a naturally-occurring immunoglobulin region and typically includes cysteine residues linking two heavy chains via disulfide bonds as in natural immunoglobulins. Representative sequences of hinge regions for human and mouse immunoglobulin can be found in ANTIBODY ENGINEERING, a PRACTICAL GUIDE, (Borrebaeck, ed., W. H. Freeman and Co., 1992).
Suitable hinge regions for the present invention can be derived from IgGI, IgG2, IgG3, IgG4, and other immunoglobulin isotypes. The IgGI isotype has two disulfide bonds in the hinge region permitting efficient and consistent disulfide bonding formation.
Therefore, a preferred hinge region of the present invention is derived from IgGI.
Optionally, the first, most N-terminal cysteine of an IgGI hinge is mutated to enhance the expression and assembly of antibodies or antibody fusion proteins of the invention (see, e.g., U.S. patent application 10/093,958, published as U.S. patent application publication 2003-0044423-Al).
In contrast to IgGI, the hinge region of IgG4 is known to form interchain disulfide bonds inefficiently (Angal et al., (1993), Mol. Immuriol. 30:105-8). Also, the IgG2 hinge region has four disulfide bonds that tend to promote oligomerization and possibly incorrect disulfide bonding during secretion in recombinant systems. One suitable hinge region for the present invention can be derived from the IgG4 hinge region, preferentially containing a mutation that enhances correct formation of disulfide bonds between heavy chain-derived moieties (Angal et al., (1993), Mol. Immunol. 30(1):105-8).
Another preferred hinge region is derived from an IgG2 hinge in which the first two cysteines are each mutated to another amino acid, such as, in order of general preference, serine, alanine, threonine, proline, glutamic acid, glutamine, lysine, histidine, arginine, asparagine, aspartic acid, glycine, methionine, valine, isoleucine, leucine, tyrosine, phenylalanine, tryptophan or selenocysteine (see, e.g., U.S. patent application publication 2003-0044423-Al).
Ii. Fc portion The antibody variable domains of the present invention are optionally fused to an Fc portion. As used herein, the Fe portion encompasses domains derived from the heavy chain constant region of an immunoglobulin, preferably a human immunoglobulin, including a fragment, analog, variant, mutant or derivative of the constant region. The constant region of an immunoglobulin heavy chain is defined as a naturally-occurring or synthetically produced polypeptide homologous to at least a portion of the C-terminal region of the heavy chain, including the CH1, hinge, CH2, CH3, and, for some heavy chain classes, CH4 domains. The "hinge" region joins the CH1 domain to the CH2-region of an Fc portion. The constant region of the heavy chains of all mammalian immunoglobulins exhibit extensive amino acid sequence similarity. DNA
sequences for these immunoglobulin regions are well known in the art. (See, e.g., Gillies et al. (1989) J.
Immunol. Meth. 125:191).
In the present invention, the Fc portion typically includes at least a CH2 domain.
For example, the Fc portion can include the entire immunoglobulin heavy chain constant region (CHl-hinge-CH2-CH3). Alternatively, the Fc portion can include all or a portion of the hinge region, the CH2 domain and the CH3 domain.
The constant region of an immunoglobulin is responsible for many important antibody effector functions, including Fc receptor (FcR) binding and complement fixation. There are five major classes of the heavy chain constant region, classified as IgA, IgG, IgD, IgE, and IgM, each with characteristic effector functions designated by isotype.
IgG, for example, is separated into four y isotypes: yl, y2, y3, and y4, also known as IgGl, IgG2, IgG3, and IgG4, respectively. IgG molecules can interact with multiple classes of cellular receptors including three classes of Fcy receptors (FcyR) specific for the IgG class of antibody, namely FcyRI, FcyRII, and FcyRIII. The sequences important for the binding of IgG to the FcyR receptors have been reported to be in the CH2 and CH3 domains.
The serum half-life of an antibody is influenced by the ability of that antibody to bind to an Fc receptor (EcR). Similarly, the serum half-life of immunoglobulin fusion proteins is also influenced by the inability to bind to such receptors (Gillies et al., Cancer Research (1999) 59:2159-66). The CH2 and CH3 domains of IgG2 and IgG4 have undetectable or reduced binding affinity to Fe receptors compared to those of IgGl.
Accordingly, the serum half-life of the featured antibody can be increased by using the CH2 and/or CH3 domain from IgG2 or IgG4 isotypes. Alternatively, the antibody can include a CH2 and/or CH3 domain from IgGI or IgG3 with modification in one or more amino acids in these domains to reduce the binding affinity for Fc receptors (see, e.g., U.S. patent application 09/256,156, published as U.S. patent application publication 2003-0105294-Al).
The hinge region of the Fc portion normally adjoins the C-terminus of the CHI
domain of the heavy chain constant region. When included in the proteins of the present invention, the hinge is homologous to a naturally-occurring immunoglobulin region and typically includes cysteine residues linking two heavy chains via disulfide bonds as in natural immunoglobulins. Representative sequences of hinge regions for human and mouse immunoglobulin can be found in ANTIBODY ENGINEERING, a PRACTICAL GUIDE, (Borrebaeck, ed., W. H. Freeman and Co., 1992).
Suitable hinge regions for the present invention can be derived from IgGI, IgG2, IgG3, IgG4, and other immunoglobulin isotypes. The IgGI isotype has two disulfide bonds in the hinge region permitting efficient and consistent disulfide bonding formation.
Therefore, a preferred hinge region of the present invention is derived from IgGI.
Optionally, the first, most N-terminal cysteine of an IgGI hinge is mutated to enhance the expression and assembly of antibodies or antibody fusion proteins of the invention (see, e.g., U.S. patent application 10/093,958, published as U.S. patent application publication 2003-0044423-Al).
In contrast to IgGI, the hinge region of IgG4 is known to form interchain disulfide bonds inefficiently (Angal et al., (1993), Mol. Immuriol. 30:105-8). Also, the IgG2 hinge region has four disulfide bonds that tend to promote oligomerization and possibly incorrect disulfide bonding during secretion in recombinant systems. One suitable hinge region for the present invention can be derived from the IgG4 hinge region, preferentially containing a mutation that enhances correct formation of disulfide bonds between heavy chain-derived moieties (Angal et al., (1993), Mol. Immunol. 30(1):105-8).
Another preferred hinge region is derived from an IgG2 hinge in which the first two cysteines are each mutated to another amino acid, such as, in order of general preference, serine, alanine, threonine, proline, glutamic acid, glutamine, lysine, histidine, arginine, asparagine, aspartic acid, glycine, methionine, valine, isoleucine, leucine, tyrosine, phenylalanine, tryptophan or selenocysteine (see, e.g., U.S. patent application publication 2003-0044423-Al).
An Fc portion fused to an antibody variable region of the invention can contain CH2 and/or CH3 domains and a hinge region that are derived from different antibody isotypes. For example, the Fc portion can contain CH2 and/or CH3 domains of IgG2 or IgG4 and a hinge region of IgGl. Assembly of such hybrid Fe portions has been described in U.S. patent application publication 2003-0044423-Al.
When fused to an antibody variable region of the invention, the Fc portion preferably contains one or more amino acid modifications that generally extend the serum half-life of an Fc fusion protein. Such amino acid modifications include mutations substantially decreasing or eliminating Fc receptor binding or complement fixing activity.
For example, one type of such mutation removes the glycosylation site of the Fc portion of an immunoglobulin heavy chain. In IgGl, the glycosylation site is Asn297 (see, for example, U.S. patent application 10/310,719, published as U.S. patent application publication 2003-0166163-Al).
III. Fusion junction region The antibody variable regions of the present invention can optionally be linked or fused to a non-immunoglobulin moiety directly or indirectly, such as through a linker peptide (e.g., (Gly4-Ser)3 (SEQ ID NO: 3)). The immunogenicity of the disclosed fusion proteins can be reduced by impairing the ability of the fusion junction or junctional epitope to interact with a T-cell receptor, as described in U. S. patent application publication 2003-0166877-Al. Even in a fusion between two human proteins, e.g., human Fc and human IL-2, the region surrounding the fusion junction or junctional epitope includes a peptide sequence that is not normally present in the human body and, thus, that can be immunogenic. The immunogenicity of the junctional epitope can be reduced, for example, by introducing one or more glycosylation sites near the fusion junction, or by identifying a candidate T-cell epitope spanning the junction as described in U.S. patent application publication 2003-0166877-Al and changing an amino acid near the junction to reduce the ability of the candidate T-cell epitope to interact with a T-cell receptor.
The serum half-life of the protein can also be increased by introducing mutations into the fusion junction region. For example, in a protein including a CH3 domain fused to a non-immunoglobulin moiety, the C-terminal lysine of the CH3 domain can be changed to another amino acid, such as alanine, which can provide a substantial increase in serum half-life of the resulting fusion protein.
When fused to an antibody variable region of the invention, the Fc portion preferably contains one or more amino acid modifications that generally extend the serum half-life of an Fc fusion protein. Such amino acid modifications include mutations substantially decreasing or eliminating Fc receptor binding or complement fixing activity.
For example, one type of such mutation removes the glycosylation site of the Fc portion of an immunoglobulin heavy chain. In IgGl, the glycosylation site is Asn297 (see, for example, U.S. patent application 10/310,719, published as U.S. patent application publication 2003-0166163-Al).
III. Fusion junction region The antibody variable regions of the present invention can optionally be linked or fused to a non-immunoglobulin moiety directly or indirectly, such as through a linker peptide (e.g., (Gly4-Ser)3 (SEQ ID NO: 3)). The immunogenicity of the disclosed fusion proteins can be reduced by impairing the ability of the fusion junction or junctional epitope to interact with a T-cell receptor, as described in U. S. patent application publication 2003-0166877-Al. Even in a fusion between two human proteins, e.g., human Fc and human IL-2, the region surrounding the fusion junction or junctional epitope includes a peptide sequence that is not normally present in the human body and, thus, that can be immunogenic. The immunogenicity of the junctional epitope can be reduced, for example, by introducing one or more glycosylation sites near the fusion junction, or by identifying a candidate T-cell epitope spanning the junction as described in U.S. patent application publication 2003-0166877-Al and changing an amino acid near the junction to reduce the ability of the candidate T-cell epitope to interact with a T-cell receptor.
The serum half-life of the protein can also be increased by introducing mutations into the fusion junction region. For example, in a protein including a CH3 domain fused to a non-immunoglobulin moiety, the C-terminal lysine of the CH3 domain can be changed to another amino acid, such as alanine, which can provide a substantial increase in serum half-life of the resulting fusion protein.
In certain embodiments, proteolytic cleavage of the fusion junction is desirable.
Accordingly, the intergenic region can include a nucleotide sequence encoding a proteolytic cleavage site. This site, interposed between the immunoglobulin and the cytokine, can be designed to provide for proteolytic release of the cytokine at the target site. For example, it is well known that plasmin and trypsin cleave after lysine and arginine residues at sites that are accessible to the proteases. Other site-specific endoproteases and the amino acid sequences they recognize are well-known.
IV. Treatment of human disease with hu14.18 antibody fusion proteins The antibody variable regions of the invention can be attached to a diagnostic and/or a therapeutic agent. The agent can be fused to the antibody to produce a fusion protein. Alternatively, the agent can be chemically coupled to the antibody to produce an immuno-conjugate. The agent can be, for example, a toxin, radiolabel, imaging agent, immunostimulatory moiety or the like.
The antibody variable region of the invention can be attached to a cytokine.
Preferred cytokines include interleukins such as interleukin-2 (IL-2), IL-4, IL-5, IL-6, IL-7, IL-10, IL-12, IL-13, IL-14, IL-15, IL-16 and IL-18, hematopoietic factors such as granulocyte-macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF) and erythropoeitin, tumor necrosis factors (TNF) such as TNF , lymphokines such as lymphotoxin, regulators of metabolic processes such as leptin, interferons such as interferon a, interferon B, and interferon y and chemokines.
Preferably, the antibody-cytokine fusion protein or immunoconjugate displays cytokine biological activity. In one embodiment, the antibody variable domain is fused to IL-2.
Preferably, several amino acids within the IL-2 moiety are mutated to reduce toxicity, as described in U.S. patent application publication 2003-0166163-Al.
For example, FIGS. 3A and 3B show the amino acid sequences of a particular embodiment of an antibody fusion protein in accordance with the invention.
Specifically, FIG. 3A shows the peptide sequence of a humanized immunoglobulin light chain that includes a variable and constant region. FIG. 3B shows the peptide sequence of a humanized immunoglobulin heavy chain linked to IL-2. The polypeptides provide a humanized antibody fusion protein capable of specifically binding to GD2 and stimulating the immune system.
Optionally, the protein complexes can further include a second agent, such as a second cytokine. In one embodiment, a hul4.18 antibody fusion protein includes and IL-2. The construction of protein complexes containing an immunoglobulin domain and two, different cytokines is described in detail in U.S. Patent No.
6,617,135.
Fusion proteins of the present invention are useful in treating human disease, such as cancer. When treating human tumors, it is particularly useful to administer an antibody-IL-2 fusion protein comprising the V regions of the invention by infusion or subcutaneous injection, using doses of 0.1 to 100 milligrams/meter2/patient.
In a preferred embodiment, it is particularly useful to administer an antibody-IL-2 fusion protein comprising the V regions of the invention by infusion or subcutaneous injection, using doses of 1 to 10 milligrams/meter2/patient, and more preferably about 3 to 6 milligrams/meter2/patient.
Clinical studies have shown that following administration of hul4.18-IL-2, the fusion protein retains its ability to activate IL-2 responsive cells through the IL-2 receptor and retains its ability to bind to GD2-positive tumor cells and to deliver IL-2 to their surface. Furthermore, administration of hul4.18-IL-2 fusion protein to a cancer patients resulted in stabilization of disease progression in a surprisingly large number of patients (see Example 1).
Pharmaceutical compositions of the invention may be used in the form of solid, semisolid, or liquid dosage forms, such as, for example, pills, capsules, powders, liquids, suspensions, or the like, preferably in unit dosage forms suitable for administration of precise dosages. The compositions include a conventional pharmaceutical carrier or excipient and, in addition, may include other medicinal agents, pharmaceutical agents, carriers, adjuvants, etc. Such excipients may include other proteins, such as, for example, human serum albumin or plasma proteins. Actual methods of preparing such dosage forms are known or will be apparent to those skilled in the art. The composition or formulation to be administered will, in any event, contain a quantity of the active component(s) in an amount effective to achieve the desired effect in the subject being treated.
Administration of the compositions hereof can be via any of the accepted modes of administration for agents that exhibit such activity. These methods include oral, parenteral, or topical administration and otherwise systemic forms.
Intravenous injection in a pharmaceutically acceptable carrier is a preferred method of administration (see Example 1).
Accordingly, the intergenic region can include a nucleotide sequence encoding a proteolytic cleavage site. This site, interposed between the immunoglobulin and the cytokine, can be designed to provide for proteolytic release of the cytokine at the target site. For example, it is well known that plasmin and trypsin cleave after lysine and arginine residues at sites that are accessible to the proteases. Other site-specific endoproteases and the amino acid sequences they recognize are well-known.
IV. Treatment of human disease with hu14.18 antibody fusion proteins The antibody variable regions of the invention can be attached to a diagnostic and/or a therapeutic agent. The agent can be fused to the antibody to produce a fusion protein. Alternatively, the agent can be chemically coupled to the antibody to produce an immuno-conjugate. The agent can be, for example, a toxin, radiolabel, imaging agent, immunostimulatory moiety or the like.
The antibody variable region of the invention can be attached to a cytokine.
Preferred cytokines include interleukins such as interleukin-2 (IL-2), IL-4, IL-5, IL-6, IL-7, IL-10, IL-12, IL-13, IL-14, IL-15, IL-16 and IL-18, hematopoietic factors such as granulocyte-macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF) and erythropoeitin, tumor necrosis factors (TNF) such as TNF , lymphokines such as lymphotoxin, regulators of metabolic processes such as leptin, interferons such as interferon a, interferon B, and interferon y and chemokines.
Preferably, the antibody-cytokine fusion protein or immunoconjugate displays cytokine biological activity. In one embodiment, the antibody variable domain is fused to IL-2.
Preferably, several amino acids within the IL-2 moiety are mutated to reduce toxicity, as described in U.S. patent application publication 2003-0166163-Al.
For example, FIGS. 3A and 3B show the amino acid sequences of a particular embodiment of an antibody fusion protein in accordance with the invention.
Specifically, FIG. 3A shows the peptide sequence of a humanized immunoglobulin light chain that includes a variable and constant region. FIG. 3B shows the peptide sequence of a humanized immunoglobulin heavy chain linked to IL-2. The polypeptides provide a humanized antibody fusion protein capable of specifically binding to GD2 and stimulating the immune system.
Optionally, the protein complexes can further include a second agent, such as a second cytokine. In one embodiment, a hul4.18 antibody fusion protein includes and IL-2. The construction of protein complexes containing an immunoglobulin domain and two, different cytokines is described in detail in U.S. Patent No.
6,617,135.
Fusion proteins of the present invention are useful in treating human disease, such as cancer. When treating human tumors, it is particularly useful to administer an antibody-IL-2 fusion protein comprising the V regions of the invention by infusion or subcutaneous injection, using doses of 0.1 to 100 milligrams/meter2/patient.
In a preferred embodiment, it is particularly useful to administer an antibody-IL-2 fusion protein comprising the V regions of the invention by infusion or subcutaneous injection, using doses of 1 to 10 milligrams/meter2/patient, and more preferably about 3 to 6 milligrams/meter2/patient.
Clinical studies have shown that following administration of hul4.18-IL-2, the fusion protein retains its ability to activate IL-2 responsive cells through the IL-2 receptor and retains its ability to bind to GD2-positive tumor cells and to deliver IL-2 to their surface. Furthermore, administration of hul4.18-IL-2 fusion protein to a cancer patients resulted in stabilization of disease progression in a surprisingly large number of patients (see Example 1).
Pharmaceutical compositions of the invention may be used in the form of solid, semisolid, or liquid dosage forms, such as, for example, pills, capsules, powders, liquids, suspensions, or the like, preferably in unit dosage forms suitable for administration of precise dosages. The compositions include a conventional pharmaceutical carrier or excipient and, in addition, may include other medicinal agents, pharmaceutical agents, carriers, adjuvants, etc. Such excipients may include other proteins, such as, for example, human serum albumin or plasma proteins. Actual methods of preparing such dosage forms are known or will be apparent to those skilled in the art. The composition or formulation to be administered will, in any event, contain a quantity of the active component(s) in an amount effective to achieve the desired effect in the subject being treated.
Administration of the compositions hereof can be via any of the accepted modes of administration for agents that exhibit such activity. These methods include oral, parenteral, or topical administration and otherwise systemic forms.
Intravenous injection in a pharmaceutically acceptable carrier is a preferred method of administration (see Example 1).
The amount of active compound administered will, of course, be dependent on the subject being treated, the severity of the affliction, the manner of administration, and the judgment of the prescribing physician.
Nucleic acids of the invention I. hul4.18 antibody constructs The invention also features nucleic acids capable of expressing each of the above types of proteins. These include, for example, nucleic acids encoding the amino acid sequence set forth in SEQ ID NO: 1; the amino acid sequence set forth in SEQ
ID NO: 2;
a hu14.18 antibody VL region that includes the huVLFR1 amino acid sequence; a hul4.18 antibody VH region that includes the huVHFR1 amino acid sequence; a hul4.18 antibody VH region that includes huVHFR3 amino acid sequence; and fusion proteins comprising a hu14.18 antibody including at least one of the foregoing humanized FR
sequences and one or more therapeutic agents.
The hul4.18 antibodies of this invention can be produced by genetic engineering techniques; i.e., by forming a nucleic acid construct encoding an GD2 specific antibody containing the desired FRs of the present invention. In one embodiment, the gene construct encoding the featured antibody includes, in 5' to 3' orientation, a DNA segment which encodes a heavy chain variable region including at least one humanized FR therein and a DNA segment encoding a heavy chain constant region. In another embodiment, another DNA segment encoding a cytokine is fused to the 3' end of the DNA
segment encoding the heavy chain constant region. In a different embodiment, the gene construct includes, in 5' to 3' orientation, a DNA segment encoding a heavy chain variable region including at least one humanized FR and a DNA segment encoding a cytokine.
Alternatively, a nucleic acid of the invention can include, in 5' to 3' orientation, a DNA
segment encoding a light chain variable region including at least one humanized FR
therein and a DNA segment encoding a cytokine. In some embodiments, a nucleic acid encoding a cytokine is joined in frame to the 3' end of a gene encoding a constant region (e.g., CH3 exon), either directly or through an intergenic region (e.g., by appropriate linkers, such as by DNA encoding (Gly4 -Ser)3 (SEQ ID NO: 3)).
II. Expression of hul4.18 antibody constructs Nucleic acid encoding proteins of the present invention can be assembled or inserted into one or more expression vectors for introduction into an appropriate recipient cell where it is expressed. The introduction of nucleic acids into expression vectors can be accomplished by standard molecular biology techniques. Preferred expression vectors include those from which the encoded protein can be expressed in either bacteria or mammalian cells.
In accordance with the invention, a heavy chain of an antibody variable region is preferably co-expressed in the same cell with a corresponding light chain. For fusion proteins that comprise multiple polypeptide chains, more than one expression vector can be used. Co-transfection methods using, for example, two expression vectors, frequently result in both vectors being delivered to a target cell. Alternatively, it is sometimes useful to use a single vector encoding a plurality of polypeptides for co-expression in the same cell.
For example, FIGS. 2A-D show the nucleic acid sequence of a single vector encoding both the heavy and light chains of an immunoglobulin in accordance with the invention. The vector also includes a nucleic acid encoding IL-2 fused to the 3' end of the immunoglobulin heavy chain. Thus, when introduced into a cell, this vector alone can provide a humanized antibody-IL-2 fusion protein that specifically binds GD2 and stimulates immune function.
Furthermore, it can be convenient to express the proteins of the present invention as single-chain molecules. For example, an antibody variable region can be expressed as a single chain antibody or sFv optionally fused to a non-immunoglobulin protein. In another embodiment, a heavy chain (with or without a fused cytokine) is combined with a light (or heavy) chain counterpart (with or without a fused cytokine) to form monovalent and divalent immunoconjugates.
Recipient cell lines are preferably lymphoid cells, such as a myeloma (or hybridoma). Myelomas can synthesize, assemble, and secrete immunoglobulins encoded by transfected genes and can glycosylate proteins. A particularly preferred recipient cell is the Sp2/0 myeloma, which normally does not produce endogenous immunoglobulin.
When transfected, the cell will produce only immunoglobulins encoded by the transfected gene constructs. Transfected myelomas can be grown in culture or in the peritonea of mice where secreted immunoconjugates can be recovered from ascites fluid.
Other lymphoid cells such as B lymphocytes can also be used as recipient cells.
There are several methods for transfectiog lymphoid cells with vectors containing the nucleic acid constructs encoding the chimeric Ig chain. A preferred way of introducing a vector into lymphoid cells is by spheroblast fusion. (see, e.g., Gillies et al.
(1989) Biotechnol. 7:798-804). Alternative methods include electroporation or calcium phosphate precipitation. Other useful methods of producing the immunoconjugates include the preparation of an RNA sequence encoding the construct and its translation in an appropriate in vivo or in vitro system. Once expressed, the proteins of the invention can be harvested by standard protein purification procedures (see, e.g , U.S.
Patent No.
5,650,150).
M. Treatment of cancer bygene therapy The nucleic acids of the invention can be used as gene therapy agents for treatment of cancer and other diseases in which it is desirable to target the immune system to a specific cell type. For example, cells can be withdrawn from a human or animal, and one or more nucleic acids encoding an antibody of the present invention can be transfected into the cells. The cells are then reintroduced into the human or animal. The transfected cells can be normal or cancer cells. Alternatively, a nucleic acid can be introduced into cells in situ. The human or animal then mounts an immune response to the cancer cells, which*can cure or lessen the severity of the cancer. An antibody variable region of the invention, coupled to appropriate regulatory elements to promote expression in mammalian cells, can be transfected into the cells by any of a variety of techniques, including via calcium phosphate, a "gene gun", adenovirus vectors, cationic liposomes, retroviral vectors, or any other efficient transfection method.
In a particular embodiment of the invention, a hu14.18 antibody is used to selectively deliver a cytokine to a target cell in vivo so that the cytokine can exert a localized biological effect such as a local inflammatory response, stimulation of T cell growth and activation, or ADCC activity. A therapeutically effective amount of the antibody is administered into the circulatory system of a subject harboring the target cell.
The invention is illustrated further by the non-limiting examples.
EXAMPLES
Example 1 .
Purification and formulation of hul4.18-IL2 In one study, hul4.18-IL2 was expressed from NS/O cells, tissue culture supernatant was harvested, and the hul4.18-1L2 protein was purified using, in sequence, Abx Mixed Resin column chromatography, recombinant Protein A chromatography, and TM TM
Q Sepharose column chromatography, followed by Pellicon 2 tangential flow diafiitration for buffer exchange into formulation buffer. Details of these purification steps are described below. Virus inactivation and removal steps were interdigitated into these steps as described below. The virus inactivation and removal steps were not necessary for purification per se, but were used to satisfy regulatory considerations.
Two liters of NS/0 tissue culture supernatant containing hul4.18-IL2 was pH-adjusted to 5.9 with 1M acetic acid and was applied to an Abx column (J. T.
Baker);
washed with 10 mM MES, 100 mM sodium acetate pH 6.2; and eluted with 500 mM
sodium acetate pH 7. This material was loaded onto a recombinant Protein A
column (Pharmacia); washed with 100 mM sodium phosphate, 150 mM NaCl pH 7; washed with 100 mM sodium phosphate, 150 mM NaCl pH 6; washed with 10 mM sodium phosphate pH 7; and eluted with 100 mM sodium phosphate, 150 mM NaCl pH 3.5. The pH of the eluted material was 4.2. To promote virus inactivation, this pH was reduced to 3.8 and the preparation was incubated for 30 minutes, after which the pH was neutralized to 7 with 1M NaOH. To remove nucleic acid, this material was loaded onto a Q
sepharose column (Pharmacia) and washed with 100 mM sodium phosphate, 150 mM NaCl pH 7.
Nucleic acid bound to the column, while the protein was found in the flow through and washes, which were repeated until the A280 returned to baseline. Pellicon 2 diafiltration (Millipore) was performed according to the manufacturer's instructions, so that the final hul4.18-IL2 material was placed in the following formulation.
24. Mannitol 4%
2 Arginine Hydrochloride USP/NF 100 mm 3. Citric Acid USP-FCC 5 mM
4. Polysorbate 80. 0.01% (w.v) The pH of the formulation buffer was adjusted to 7 with 1 M NaOH.
As a final step, the preparation was filtered through a Viresolve 180 membrane (Millipore), which has a molecular weight cutoff of 180,000 Daltons. This had the effect of `polishing' the material so that as a result, aggregated dimers and higher-order oligomers were removed.
Example 2 Anti-tumor Activity of the hul4.18-IL-2 Fusion Protein Observed in Phase I Clinical Trials To evaluate the safety and efficacy of hul4.18-IL-2, a Phase I clinical trial was performed. Eligible patients had histologically confirmed melanoma that was considered surgically and medically incurable. These patients could have either measurable or evaluable metastatic disease, or they could have no evidence of disease following surgical resection of either distant metastases or regionally recurrent disease.
Patients with multiple (two or more) local or regional recurrences were included only if they had prior evidence of lymph node involvement and if each recurrence was separated in time by at least 2 months. All patients needed to have adequate bone marrow function (defined by total white blood cells (WBC) > 3,500/ml, or total granulocytes > 2000/ml, platelets >
100,000/ml, and hemoglobin >10.0 g/dl), adequate liver fimction [defined by an aspartate aminotransferase (AST) < 3 x normal and a total bilirubin < 2.0 mg/dl], and adequate renal function (defined by a serum creatinine <2.0 mg/dl or a creatinine clearance of >60 ml/minute). All patients had an electrocorticography (ECOG) performance status of 0 or 1 and a life expectancy of at least 12 weeks. Patients who had previously received chemotherapy, radiation therapy, or other,immunosuppressive therapy within 4 weeks prior to study were excluded. Patients could have prior central nervous system (CNS) metastases if treated and stable for at least 4 weeks prior to starting the study. Informed consent was obtained from all patients.
This phase I trial was designed as an open-label, nonrandomized dose escalation study in which groups of 3 to 6 patients received hul4.18-IL-2 at one of the following dose levels: 0.8, 1.6, 3.2, 4.8, 6.0 or 7.5 mg/m2/day. The hul4.18-IL-2 was administered on an inpatient basis as a 4-hour intravenous (IV) infusion over 3 consecutive days during the first week of each course. The hu14.18-IL-2 fusion protein was administered to patients in a formulation comprising 4% Mannitol; Arginine HCl, 100 mM;
Citrate, 5 TM
mM; and 0.01% Tween 80, at pH 7. Patients were discharged from the hospital, if stable, approximately 24 hours following the completion of the third infusion. Adverse events and toxicities were graded as per NCI Common Toxicity Criteria (version 2.0) and the University of Wisconsin Comprehensive Cancer Center Toxicity Grading Scale for (performance status, weight gain, and temperature). Dose-limiting toxicity (DLT) was defined as the occurrence of grade 3 or 4 toxicity other than grade 3 lymphopenia, hyperbilirubinemia, hypophosphatemia or hyperglycemia. The maximal tolerated dose (MTD) was defined as the dose level at which two of six patients had DLT
during course 1. Patients with grade 3 treatment-related toxicities were required to recover to at least grade 1 before they could resume treatment at a 50% dose reduction for course 2.
Patients with >25% disease progression were removed from the study. Patients with stable disease were administered course 2.
Nucleic acids of the invention I. hul4.18 antibody constructs The invention also features nucleic acids capable of expressing each of the above types of proteins. These include, for example, nucleic acids encoding the amino acid sequence set forth in SEQ ID NO: 1; the amino acid sequence set forth in SEQ
ID NO: 2;
a hu14.18 antibody VL region that includes the huVLFR1 amino acid sequence; a hul4.18 antibody VH region that includes the huVHFR1 amino acid sequence; a hul4.18 antibody VH region that includes huVHFR3 amino acid sequence; and fusion proteins comprising a hu14.18 antibody including at least one of the foregoing humanized FR
sequences and one or more therapeutic agents.
The hul4.18 antibodies of this invention can be produced by genetic engineering techniques; i.e., by forming a nucleic acid construct encoding an GD2 specific antibody containing the desired FRs of the present invention. In one embodiment, the gene construct encoding the featured antibody includes, in 5' to 3' orientation, a DNA segment which encodes a heavy chain variable region including at least one humanized FR therein and a DNA segment encoding a heavy chain constant region. In another embodiment, another DNA segment encoding a cytokine is fused to the 3' end of the DNA
segment encoding the heavy chain constant region. In a different embodiment, the gene construct includes, in 5' to 3' orientation, a DNA segment encoding a heavy chain variable region including at least one humanized FR and a DNA segment encoding a cytokine.
Alternatively, a nucleic acid of the invention can include, in 5' to 3' orientation, a DNA
segment encoding a light chain variable region including at least one humanized FR
therein and a DNA segment encoding a cytokine. In some embodiments, a nucleic acid encoding a cytokine is joined in frame to the 3' end of a gene encoding a constant region (e.g., CH3 exon), either directly or through an intergenic region (e.g., by appropriate linkers, such as by DNA encoding (Gly4 -Ser)3 (SEQ ID NO: 3)).
II. Expression of hul4.18 antibody constructs Nucleic acid encoding proteins of the present invention can be assembled or inserted into one or more expression vectors for introduction into an appropriate recipient cell where it is expressed. The introduction of nucleic acids into expression vectors can be accomplished by standard molecular biology techniques. Preferred expression vectors include those from which the encoded protein can be expressed in either bacteria or mammalian cells.
In accordance with the invention, a heavy chain of an antibody variable region is preferably co-expressed in the same cell with a corresponding light chain. For fusion proteins that comprise multiple polypeptide chains, more than one expression vector can be used. Co-transfection methods using, for example, two expression vectors, frequently result in both vectors being delivered to a target cell. Alternatively, it is sometimes useful to use a single vector encoding a plurality of polypeptides for co-expression in the same cell.
For example, FIGS. 2A-D show the nucleic acid sequence of a single vector encoding both the heavy and light chains of an immunoglobulin in accordance with the invention. The vector also includes a nucleic acid encoding IL-2 fused to the 3' end of the immunoglobulin heavy chain. Thus, when introduced into a cell, this vector alone can provide a humanized antibody-IL-2 fusion protein that specifically binds GD2 and stimulates immune function.
Furthermore, it can be convenient to express the proteins of the present invention as single-chain molecules. For example, an antibody variable region can be expressed as a single chain antibody or sFv optionally fused to a non-immunoglobulin protein. In another embodiment, a heavy chain (with or without a fused cytokine) is combined with a light (or heavy) chain counterpart (with or without a fused cytokine) to form monovalent and divalent immunoconjugates.
Recipient cell lines are preferably lymphoid cells, such as a myeloma (or hybridoma). Myelomas can synthesize, assemble, and secrete immunoglobulins encoded by transfected genes and can glycosylate proteins. A particularly preferred recipient cell is the Sp2/0 myeloma, which normally does not produce endogenous immunoglobulin.
When transfected, the cell will produce only immunoglobulins encoded by the transfected gene constructs. Transfected myelomas can be grown in culture or in the peritonea of mice where secreted immunoconjugates can be recovered from ascites fluid.
Other lymphoid cells such as B lymphocytes can also be used as recipient cells.
There are several methods for transfectiog lymphoid cells with vectors containing the nucleic acid constructs encoding the chimeric Ig chain. A preferred way of introducing a vector into lymphoid cells is by spheroblast fusion. (see, e.g., Gillies et al.
(1989) Biotechnol. 7:798-804). Alternative methods include electroporation or calcium phosphate precipitation. Other useful methods of producing the immunoconjugates include the preparation of an RNA sequence encoding the construct and its translation in an appropriate in vivo or in vitro system. Once expressed, the proteins of the invention can be harvested by standard protein purification procedures (see, e.g , U.S.
Patent No.
5,650,150).
M. Treatment of cancer bygene therapy The nucleic acids of the invention can be used as gene therapy agents for treatment of cancer and other diseases in which it is desirable to target the immune system to a specific cell type. For example, cells can be withdrawn from a human or animal, and one or more nucleic acids encoding an antibody of the present invention can be transfected into the cells. The cells are then reintroduced into the human or animal. The transfected cells can be normal or cancer cells. Alternatively, a nucleic acid can be introduced into cells in situ. The human or animal then mounts an immune response to the cancer cells, which*can cure or lessen the severity of the cancer. An antibody variable region of the invention, coupled to appropriate regulatory elements to promote expression in mammalian cells, can be transfected into the cells by any of a variety of techniques, including via calcium phosphate, a "gene gun", adenovirus vectors, cationic liposomes, retroviral vectors, or any other efficient transfection method.
In a particular embodiment of the invention, a hu14.18 antibody is used to selectively deliver a cytokine to a target cell in vivo so that the cytokine can exert a localized biological effect such as a local inflammatory response, stimulation of T cell growth and activation, or ADCC activity. A therapeutically effective amount of the antibody is administered into the circulatory system of a subject harboring the target cell.
The invention is illustrated further by the non-limiting examples.
EXAMPLES
Example 1 .
Purification and formulation of hul4.18-IL2 In one study, hul4.18-IL2 was expressed from NS/O cells, tissue culture supernatant was harvested, and the hul4.18-1L2 protein was purified using, in sequence, Abx Mixed Resin column chromatography, recombinant Protein A chromatography, and TM TM
Q Sepharose column chromatography, followed by Pellicon 2 tangential flow diafiitration for buffer exchange into formulation buffer. Details of these purification steps are described below. Virus inactivation and removal steps were interdigitated into these steps as described below. The virus inactivation and removal steps were not necessary for purification per se, but were used to satisfy regulatory considerations.
Two liters of NS/0 tissue culture supernatant containing hul4.18-IL2 was pH-adjusted to 5.9 with 1M acetic acid and was applied to an Abx column (J. T.
Baker);
washed with 10 mM MES, 100 mM sodium acetate pH 6.2; and eluted with 500 mM
sodium acetate pH 7. This material was loaded onto a recombinant Protein A
column (Pharmacia); washed with 100 mM sodium phosphate, 150 mM NaCl pH 7; washed with 100 mM sodium phosphate, 150 mM NaCl pH 6; washed with 10 mM sodium phosphate pH 7; and eluted with 100 mM sodium phosphate, 150 mM NaCl pH 3.5. The pH of the eluted material was 4.2. To promote virus inactivation, this pH was reduced to 3.8 and the preparation was incubated for 30 minutes, after which the pH was neutralized to 7 with 1M NaOH. To remove nucleic acid, this material was loaded onto a Q
sepharose column (Pharmacia) and washed with 100 mM sodium phosphate, 150 mM NaCl pH 7.
Nucleic acid bound to the column, while the protein was found in the flow through and washes, which were repeated until the A280 returned to baseline. Pellicon 2 diafiltration (Millipore) was performed according to the manufacturer's instructions, so that the final hul4.18-IL2 material was placed in the following formulation.
24. Mannitol 4%
2 Arginine Hydrochloride USP/NF 100 mm 3. Citric Acid USP-FCC 5 mM
4. Polysorbate 80. 0.01% (w.v) The pH of the formulation buffer was adjusted to 7 with 1 M NaOH.
As a final step, the preparation was filtered through a Viresolve 180 membrane (Millipore), which has a molecular weight cutoff of 180,000 Daltons. This had the effect of `polishing' the material so that as a result, aggregated dimers and higher-order oligomers were removed.
Example 2 Anti-tumor Activity of the hul4.18-IL-2 Fusion Protein Observed in Phase I Clinical Trials To evaluate the safety and efficacy of hul4.18-IL-2, a Phase I clinical trial was performed. Eligible patients had histologically confirmed melanoma that was considered surgically and medically incurable. These patients could have either measurable or evaluable metastatic disease, or they could have no evidence of disease following surgical resection of either distant metastases or regionally recurrent disease.
Patients with multiple (two or more) local or regional recurrences were included only if they had prior evidence of lymph node involvement and if each recurrence was separated in time by at least 2 months. All patients needed to have adequate bone marrow function (defined by total white blood cells (WBC) > 3,500/ml, or total granulocytes > 2000/ml, platelets >
100,000/ml, and hemoglobin >10.0 g/dl), adequate liver fimction [defined by an aspartate aminotransferase (AST) < 3 x normal and a total bilirubin < 2.0 mg/dl], and adequate renal function (defined by a serum creatinine <2.0 mg/dl or a creatinine clearance of >60 ml/minute). All patients had an electrocorticography (ECOG) performance status of 0 or 1 and a life expectancy of at least 12 weeks. Patients who had previously received chemotherapy, radiation therapy, or other,immunosuppressive therapy within 4 weeks prior to study were excluded. Patients could have prior central nervous system (CNS) metastases if treated and stable for at least 4 weeks prior to starting the study. Informed consent was obtained from all patients.
This phase I trial was designed as an open-label, nonrandomized dose escalation study in which groups of 3 to 6 patients received hul4.18-IL-2 at one of the following dose levels: 0.8, 1.6, 3.2, 4.8, 6.0 or 7.5 mg/m2/day. The hul4.18-IL-2 was administered on an inpatient basis as a 4-hour intravenous (IV) infusion over 3 consecutive days during the first week of each course. The hu14.18-IL-2 fusion protein was administered to patients in a formulation comprising 4% Mannitol; Arginine HCl, 100 mM;
Citrate, 5 TM
mM; and 0.01% Tween 80, at pH 7. Patients were discharged from the hospital, if stable, approximately 24 hours following the completion of the third infusion. Adverse events and toxicities were graded as per NCI Common Toxicity Criteria (version 2.0) and the University of Wisconsin Comprehensive Cancer Center Toxicity Grading Scale for (performance status, weight gain, and temperature). Dose-limiting toxicity (DLT) was defined as the occurrence of grade 3 or 4 toxicity other than grade 3 lymphopenia, hyperbilirubinemia, hypophosphatemia or hyperglycemia. The maximal tolerated dose (MTD) was defined as the dose level at which two of six patients had DLT
during course 1. Patients with grade 3 treatment-related toxicities were required to recover to at least grade 1 before they could resume treatment at a 50% dose reduction for course 2.
Patients with >25% disease progression were removed from the study. Patients with stable disease were administered course 2.
The pharmacokinetic properties of hul4.18-IL-2 were evaluated in the patients.
When hul4.18-IL-2 levels were evaluated in serial samples from all 33 patients immediately following the first 4-hour infusion (day 1, course 1), the half-life was found to be 3.7 hours (+/- SD of 0.9 h). This is intermediate between the half-lives of its 2 components (approximately 45 minutes for IL-2 and 3 days for the chimeric m14.18 antibody), and comparable to that which was observed for the half-life of chimeric m14.18-IL-2 in mice. Following the clearance of hul4.18-IL-2 from the serum of these patients, neither the IL-2 nor hul4.18 antibody components could be detected.
The peak serum and area under the curve (AUC) during course 1 showed a significant dose-dependent increase (p<0.001).
Thirty-three patients were treated in this study. Table 1 lists clinical outcomes.
Two patients (6%) completed only the first 2 of 3 days for course 1. One of these patients (dose level 3) had a grade 3 hyperbilirubinemia on day 2 of treatment, and the other patient (dose level 6) had grade 3 hypoxia and hypotension requiring treatment to be held.
Both of these patients had progression of disease and did not receive a second course of therapy. Nineteen patients (58%) had stable disease following the first course of therapy and received a second course of therapy. Five patients (15% of all patients) required a 50% dose reduction for course 2 secondary to adverse events in course 1.
Seventeen patients (52% of all patients) completed course 2. One patient (dose level 4) declined to receive the final infusion during course 2, and one patient (dose level 6) had the final infusion during course 2 held due to hypotension. Eight patients (24% of all patients) had stable disease following the second course of treatment. The results indicate that hul4.18-IL-2 caused stabilization of disease progression in a surprisingly large number of patients.
Eight of the 33 patients maintained stable disease after 2 courses of therapy, and 4 of these 8 patients continue with no evidence of progressive disease (1 with stable disease and 3 with no evidence of disease) for 20-52 months since completing protocol therapy.
Five of the 33 patients entered the study with no measurable disease following surgical resection of recurrences or metastases. Two of these five patients had disease progression, while the remaining 3 patients continued with no evidence of disease (20-52 months). These findings are consistent with the hypothesis that clinical benefit from an immunotherapeutic intervention is most likely in a patient with a low tumor burden. One additional patient had an objective decrease in a lung nodule following two courses of therapy, but the overall disease response was scored as disease progression due to growth in a distant node. The node was resected following hul4.18-IL-2 therapy and the patient remained free from disease progression for over 3 years.
Clinical Outcomes Number of Patients Patients completing course 1 31 Stable disease following course 1 19 50% dose reduction for course 2 5 Patients completing course 2 17 Stable disease following course 2 8 Immune stimulation in vivo by hul4.18-IL-2 in a Phase I clinical trial.
Patients treated with hul4.18-IL-2 were also examined for indications of immune stimulation. A peripheral blood lymphopenia occurred on days 2-4, and this was followed by a rebound lymphocytosis on days 5-22. Both of these changes were dose-dependent (p<0.01 and p<0.05, respectively). The lymphocyte counts on days 5, 8, 15 and 22 were significantly greater than baseline for course 1. The baseline lymphocyte count for course 2 (day 29 of course 1) was increased over the baseline lymphocyte count for course 1, indicating that effects of the first course of treatment are still present on day 29. In addition, the lymphocyte counts during course 2 on days 5, 8 and 15 are greater than the corresponding values for days 5, 8, and 15 during course 1 for these 12 patients.
Lymphocyte cell surface phenotype showed an expansion of CD16+ and CD56+
lymphocytes (natural killer (NK) cell markers) following the first week of hul4.18-IL-2 therapy. This effect was still present on day 29 of course 1 (day 1, course 2). For patients 19-33 (receiving 4.8-7.5 mg/m2/day), lymphocyte cell surface phenotype was determined on days 15 and 22 in addition to days 1 and 8. This analysis demonstrated that the augmentation of CD56 and CD56/CD16 co-expressing cells remained significantly elevated (p<O.01) on days 8, 15 and 22.
As a measure of immune activation, C-reactive protein (CRP) levels for patients 13-33 and soluble IL-2 receptor (sIL-2R) levels for the 31 patients completing course 1, were obtained. A significant increase in mean CRP was present on treatment days 3-5 in both course 1 and course 2 compared to baseline for each course. This increase in CRP
returned to baseline levels by day 8 of each treatment course. The sIL-2R
level was significantly increased over baseline starting 24 hours after the hul4.18-IL-2 infusion during both course 1 and course 2, which persisted through day S. The increase in sIL-2R
was found to be dose dependent (p= 0.014). sIL-2R values for course 2 were increased compared to corresponding values in course 1 for days 1-5 for patients receiving the same dose in both courses (p < 0.05).
The LA-N-5 neuroblastoma cell line that expresses GD2 and binds hul4.18-IL-2 was used to evaluate IL-2 activated NK function and antibody dependent cellular cytotoxicity (ADCC) on peripheral blood mononuclear cells (PBMC) from 31 patients completing course 1. There was a significant increase in killing mediated by lymphocytes from day 8 'when compared with day 1 for these two assays. The 12 patients that received course 2 at the same dose as in course 1, showed ADCC results that were very similar to those obtained during course 1. The only parameter that was found to be different for course 2 from course 1 was increased killing in the presence of IL-2 on day 1, indicating that augmented killing in this assay remained elevated on day 29 (day 1, course 2).
Because the LA-N-5 target is relatively resistant to fresh NK cells, it is useful for measuring IL-2 augmented killing, and ADCC. However, the weak killing of LA-N-mediated by fresh PBMC in medium (without supplemental IL-2 in vitro) was not significantly greater on day 8 than on day 1.
For patients 19-33, standard NK assays were performed on days 1, 8, 15 and 22, using the NIA susceptible K562 target cell line. A significant increase in NK
lysis of K562 target cells, when tested either in medium or in the presence of IL-2, was observed on days 8 and 22 when compared with day 1. Serum samples from selected patients were also evaluated to determine functional IL-2 activity and functional anti-GD2 antibody.
The IL-2 responsive Tf-lb cell line demonstrated IL-2-induced proliferation with patient serum obtained following infusion of hul.4.18-IL-2. A progressive increase in proliferation was seen during the first 4 hours following the 4-hour infusion.
Values returned to baseline by 16 hours after this infusion, consistent with the serum half-life for hul4.18-IL-2 of approximately 4 hours. Serum samples from these time-points were also examined by flow cytometry for the presence of intact hul4.18-IL-2 immunocytokine (IC) that retains its IL-2 component and its anti-GD2 antibody activity.
hul4.18-IL-2 capable of binding to the M21 cell line (GD2 positive) was detectable in patient serum samples following an infusion of IC. The amount of IC able to bind to M21 progressively increased during the first 4 hours following the 4-hour infusion, and decreased after that, again consistent with the half-life of approximately 4 hours.
Finally, in vitro assays were performed with specimens from patients to determine whether administration of hul4.18-IL-2 results in conditions in vivo consistent with those needed to achieve ADCC. PBMCs from day 8 show augmented ADCC on GD2+ target cells when hul4.18-IL-2 is added to the cytotoxic assay. This same ADCC assay was performed with PBMC from day 8, however instead of adding hul4.18-IL-2 to the assay, serum from the patient, obtained before or after hul4.18-IL-2 administration, was added.
PBMC obtained from patients on day 8 of course 2 were able to mediate augmented killing of the LA-N-5 cell line in the presence of serum obtained following hul4.18-IL-2 administration, compared to that observed with serum obtained prior to infusion. Thus the hul4.18-IL-2 circulating in patients after IV administration is able to facilitate ADCC
with PBMCs activated in vivo by hul4.18-IL-2 from that same patient.
In summary, these results indicate that there were immunological changes associated with this hul4.18-IL-2 therapy including an increase in lymphocyte count, an increase in the percentage of CD16+ and CD56+ PBMC, an increase in NK lysis, and an increase in ADCC. Additional evidence for immune activation included an increase in serum levels of CRP and of sIL-2R. Laboratory analyses of serum and PBMC
showed that the hul4.18-IL-2 molecule circulating in patient serum following IV
administration retained its ability to activate IL-2 responsive cells through the IL-2 receptor and retained its ability to bind to GD2 positive tumor cells, and deliver IL-2 to their surface, as detected by flow cytometry. NK cells were activated in vivo based on their ability to mediate NK and ADCC function in vitro. Furthermore, the NK cells activated in vivo by the hul4.18-IL-2 administered to these patients were able to mediate ADCC
facilitated by the hul4.l8-IL-2 circulating in the serum of those same patients. Thus, conditions to achieve immune activation were achieved in all patients in this study.
SEQUENCE LISTING
<110> Gillies , Stephen D.
Lo, Kin-Ming <120> Immunocytokine Sequences and Uses Thereof <130> LEX-023 <150> US 60/433,945 <151> 2002-12-17 <160> 6 <170> Patentln version 3.1 <210> 1 <211> 113 <212> PRT
<213> Artificial Sequence <220>
<223> Humanized Immunoglobulin light chain variable region <400> 1 Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Arg Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gin Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile His Lys Val Ser Asn Arg Phe Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Ser Thr His Val Pro Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys <210> 2 <211> 113 <212> PRT
<213> Artificial Sequence <220>
<223> Humanized Immunoglobulin heavy chain variable region <400> 2 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Glu Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Ser Ser Phe Thr Gly Tyr Asn Met Asn Trp Val Arg Gln Asn Ile Gly Lys Ser Leu Glu Trp Ile Gly Ala Ile Asp Pro Tyr Tyr Gly Gly Thr Ser Tyr Asn Gln Lys Phe Lys Gly Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr Met His Leu Lys Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Val Ser Gly Met Glu Tyr Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser <210> 3 <211> 15 <212> PRT
<213> Artificial Sequence <220>
<223> Linker sequence <400> 3 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser <210> 4 <211> 10531 <212> DNA
<213> Artificial Sequence <220>
<223> Vector containing humanized Immunoglobulin light and heavy chain and IL-2 <400> 4 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 60 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 120 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 180 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 240 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 300 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 360 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 420 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 480 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 540 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta 600 cagaacccac tgcttaactg gcttatcgaa attaatacga ctcactatag ggagaccctc 660 tagaatgaag ttgcctgtta ggctgttggt gctgatgttc tggattcctg gtgaggagag 720 agggaagtga gggaggagaa tggacaggga gcaggagcac tgaatcccat tgctcattcc 780 atgtatctgg catgggtgag aagatgggtc ttatcctcca gcatggggcc tctggggtga 840 atacttgtta gagggaggtt ccagatggga acatgtgcta taatgaagat tatgaaatgg 900 atgcctggga tggtctaagt aatgccttag aagtgactag acacttgcaa ttcacttttt 960 ttggtaagaa gagattttta ggctataaaa aaatgttatg taaaaataaa cgatcacagt 1020 tgaaataaaa aaaaaatata aggatgttca tgaattttgt gtataactat gtatttctct 1080 ctcattgttt cagcttcctt aagcgacgtg gtgatgaccc agacccccct gtccctgccc 1140 gtgacccccg gcgagcccgc ctccatctcc tgcagatcta gtcagagtct tgtacaccgt 1200 aatggaaaca cctatttaca ttggtacctg cagaagccag gccagtctcc aaagctcctg 1260 attcacaaag tttccaaccg attttctggg gtcccagaca ggttcagtgg cagtggatca 1320 gggacagatt tcacactcaa gatcagcaga gtggaggctg aggatctggg agtttatttc 1380 tgttctcaaa gtacacatgt tcctccgctc acgttcggtg ctgggaccaa gctggagctg 1440 aaacgtatta gtgtgtcagg gtttcacaag agggactaaa gacatgtcag ctatgtgtga 1500 ctaatggtaa tgtcactaag ctgcgggatc ccgcaattct aaactctgag ggggtcggat 1560 gacgtggcca ttctttgcct aaagcattga gtttactgca aggtcagaaa agcatgcaaa 1620 gccctcagaa tggctgcaaa gagctccaac aaaacaattt agaactttat taaggaatag 1680 ggggaagcta ggaagaaact caaaacatca agattttaaa tacgcttctt ggtctccttg 1740 ctataattat ctgggataag catgctgttt tctgtctgtc cctaacatgc cctgtgatta 1800 tccgcaaaca acacacccaa gggcagaact ttgttactta aacaccatcc tgtttgcttc 1860 tttcctcagg aactgtggct gcaccatctg tcttcatctt cccgccatct gatgagcagt 1920 tgaaatctgg aactgcctct gttgtgtgcc tgctgaataa cttctatccc agagaggcca 1980 aagtacagtg gaaggtggat aacgccctcc aatcgggtaa ctcccaggag agtgtcacag 2040 agcaggacag caaggacagc acctacagcc tcagcagcac cctgacgctg agcaaagcag 2100 actacgagaa acacaaagtc tacgcctgcg aagtcaccca tcagggcctg agctcgcccg 2160 tcacaaagag cttcaacagg ggagagtgtt agagggagaa gtgcccccac ctgctcctca 2220 gttccagcct gaccccctcc catcctttgg cctctgaccc tttttccaca ggggacctac 2280 ccctattgcg gtcctccagc tcatctttca cctcaccccc ctcctcctcc ttggctttaa 2340 ttatgctaat gttggaggag aatgaataaa taaagtgaat ctttgcacct gtggtttctc 2400 tctttcctca atttaataat tattatctgt tgtttaccaa ctactcaatt tctcttataa 2460 gggactaaat atgtagtcat cctaaggcgc ataaccattt ataaaaatca tccttcattc 2520 tattttaccc tatcatcctc tgcaagacag tcctccctca aacccacaag ccttctgtcc 2580 tcacagtccc ctgggccatg gtaggagaga cttgcttcct tgttttcccc tcctcagcaa 2640 gccctcatag tcctttttaa gggtgacagg tcttacggtc atatatcctt tgattcaatt 2700 ccctgggaat caaccaaggc aaatttttca aaagaagaaa cctgctataa agagaatcat 2760 tcattgcaac atgatataaa ataacaacac aataaaagca attaaataaa caaacaatag 2820 ggaaatgttt aagttcatca tggtacttag acttaatgga atgtcatgcc ttatttacat 2880 ttttaaacag gtactgaggg actcctgtct gccaagggcc gtattgagta ctttccacaa 2940 cctaatttaa tccacactat actgtgagat taaaaacatt cattaaaatg ttgcaaaggt 3000 tctataaagc tgagagacaa atatattcta taactcagca atcccacttc tagggtcgat 3060 cgacgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt 3120 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga 3180 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca 3240 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca 3300 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg 3360 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 3420 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 3480 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 3540 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 3600 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta 3660 actacagaac ccactgctta actggcttat cgaaattaat acgactcact atagggagac 3720 ccaagctcct cgaggctaga atgaagttgc ctgttaggct gttggtgctg atgttctgga 3780 ttcctggtga ggagagaggg aagtgaggga ggagaatgga cagggagcag gagcactgaa 3840 tcccattgct cattccatgt atctggcatg ggtgagaaga tgggtcttat cctccagcat 3900 ggggcctctg gggtgaatac ttgttagagg gaggttccag atgggaacat gtgctataat 3960 gaagattatg aaatggatgc ctgggatggt ctaagtaatg ccttagaagt gactagacac 4020 ttgcaattca ctttttttgg taagaagaga tttttaggct ataaaaaaat gttatgtaaa 4080 aataaacgat cacagttgaa ataaaaaaaa aatataagga tgttcatgaa ttttgtgtat 4140 aactatgtat ttctctctca ttgtttcagc ttccttaagc gaggtgcagc tggtgcagtc 4200 cggcgccgag gtggagaagc ccggcgcctc cgtgaagatc tcctgcaagg,cctccggctc 4260 ctccttcacc ggctacaaca tgaactgggt gcgccagaac atcggcaagt ccctggagtg 4320 gatcggcgcc atcgacccct actacggcgg cacctcctac aaccagaagt tcaagggccg 4380 cgccaccctg accgtggaca agtccacctc caccgcctac atgcacctga agtccctgcg 4440 ctccgaggac accgccgtgt actactgcgt gtccggcatg gagtactggg gccagggcac 4500 ctccgtgacc gtgtcctccg gtaagctttt ctggggcagg ccaggcctga ccttggcttt 4560 ggggcaggga gggggctaag gtgaggcagg tggcgccagc caggtgcaca cccaatgccc 4620 atgagcccag acactggacg ctgaacctcg cggacagtta agaacccagg ggcctctgcg 4680 ccctgggccc agctctgtcc cacaccgcgg tcacatggca ccacctctct tgcagcctcc 4740 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca 4800 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 4860 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 4920 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 4980 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agagagttgg tgagaggcaa 5040 gcacagggag ggagggtgtc tgctggaagc caggctcagc gctcctgcct ggacgcatcc 5100 cggctatgca gtcccagtcc agggcagcaa ggcaggcccc gtctgcctct tcacccggag 5160 gcctctgccc gccccactca tgctcaggga gagggtcttc tggctttttc cccaggctct 5220 gggcaggcac aggctaggtg cccctaaccc aggccctgca cacaaagggg caggtgctgg 5280 gctcagacct gccaagagcc atatccggga ggaccctgcc cctgacctaa gcccacccca 5340 aaggccaaac tctccactcc ctcagctcgg acaccttctc tcctcccaga ttccagtaac 5400 tcccaatctt ctctctgcag agcccaaatc ttgtgacaaa actcacacat gcccaccgtg 5460 cccaggtaag ccagcccagg cctcgccctc cagctcaagg cgggacaggt gccctagagt 5520 agcctgcatc cagggacagg ccccagccgg gtgctgacac gtccacctcc atctcttcct 5580 cagcacctga actcctgggg ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca 5640 ccctcatgat ctcccggacc cctgaggtca catgcgtggt ggtggacgtg agccacgaag 5700 accctgaggt caagttcaac tggtacgtgg acggcgtgga ggtgcataat gccaagacaa 5760 agccgcggga ggagcagtac aacagcacgt accgtgtggt cagcgtcctc accgtcctgc 5820 accaggactg gctgaatggc aaggagtaca agtgcaaggt ctccaacaaa gccctcccag 5880 cccccatcga gaaaaccatc tccaaagcca aaggtgggac ccgtggggtg cgagggccac 5940 atggacagag gccggctcgg cccaccctct gccctgagag tgaccgctgt accaacctct 6000 gtccctacag ggcagccccg agaaccacag gtgtacaccc tgcccccatc acgggaggag 6060 atgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc 6120 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 6180 ctggactccg acggctcctt cttcctctat agcaagctca ccgtggacaa gagcaggtgg 6240 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 6300 cagaagagcc tctccctgtc cccgggtaaa gccccaactt caagttctac aaagaaaaca 6360 cagctgcaac tggagcatct cctgctggat ctccagatga ttctgaatgg aattaacaac 6420 tacaagaatc ccaaactcac caggatgctc acattcaagt tctacatgcc caagaaggcc 6480 acagagctca aacatctcca gtgtctagag gaggaactca aacctctgga ggaagtgcta 6540 aacctcgctc agagcaaaaa cttccactta agacctaggg acttaatcag caatatcaac 6600 gtaatagttc tggaactaaa gggatccgaa acaacattca tgtgtgaata tgctgatgag 6660 acagcaacca ttgtagaatt tctgaacaga tggattacct tttgtcaaag catcatctca 6720 acactaactt gataattaag tgctcgaggg atccagacat gataagatac attgatgagt 6780 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg 6840 ctattgcttt atttgtaacc attagaagct gcaataaaca agttaacaac aacaattgca 6900 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc 6960 tctacaaatg tggtatggct gattatgatc ctgcctcgcg cgtttcggtg atgacggtga 7020 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg 7080 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat 7140 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag 7200 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa 7260 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 7320 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 7380 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 7440 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 7500 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 7560 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 7620 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg 7680 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 7740 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 7800 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 7860 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct 7920 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 7980 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 8040 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 8100 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 8160 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 8220 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 8280 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 8340 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 8400 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 8460 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 8520 gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 8580 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 8640 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 8700 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 8760 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 8820 tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 8880 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 8940 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 9000 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 9060 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 9120 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 9180 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta 9240 acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaattccg atccagacat 9300 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt 9360 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attagaagct gcaataaaca 9420 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt 9480 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc taaagccagc 9540 aaaagtccca tggtcttata aaaatgcata gctttcggag gggagcagag aacttgaaag 9600 catcttcctg ttagtctttc ttctcgtaga ccttaaattc atacttgatt cctttttcct 9660 cctggacctc agagaggacg cctgggtatt ctgggagaag tttatatttc cccaaatcaa 9720 tttctgggaa aaacgtgtca ctttcaaatt cctgcatgat ccttgtcaca aagagtctga 9780 ggtggcctgg ttgattcatg gcttcctggt aaacagaact gcctccgact atccaaacca 9840 tgtctacttt acttgccaat tccggttgtt caataagtct taaggcatca tccaaacttt 9900 tggcaagaaa atgagctcct cgtggtggtt ctttgagttc tctactgaga actatattaa 9960 ttctgtcctt taaaggtcga ttcttctcag gaatggagaa ccaggttttc ctacccataa 10020 tcaccagatt ctgtttacct tccactgaag aggttgtggt cattctttgg aagtacttga 10080 actcgttcct gagcggaggc cagggtcggt ctccgttctt gccaatcccc atattttggg 10140 acacggcgac gatgcagttc aatggtcgaa ccatgagggc accaagctag ctttttgcaa 10200 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg 10260 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa 10320 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa 10380 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg 10440 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac 10500 tttccacacc ctaactgaca cacattccac a 10531 <210> 5 <211> 220 <212> PRT
<213> Artificial Sequence <220>
<223> Humanized Immunoglobulin light chain <400> 5 Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Arg Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile His Lys Val Ser Asn Arg Phe Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Ser Thr His Val Pro Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys <210> 6 <211> 575 <212> PRT
<213> Artificial Sequence <220>
<223> Humanized Immunoglobulin heavy chain and IL-2 <400> 6 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Glu Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Ser Ser Phe Thr Gly Tyr Asn Met Asn Trp Val Arg Gln Asn Ile Gly Lys Ser Leu Glu Trp Ile Gly Ala Ile Asp Pro Tyr Tyr Gly Gly Thr Ser Tyr Asn Gln Lys Phe Lys Gly Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr Met His Leu Lys Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Val Ser Gly Met Glu Tyr Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr
When hul4.18-IL-2 levels were evaluated in serial samples from all 33 patients immediately following the first 4-hour infusion (day 1, course 1), the half-life was found to be 3.7 hours (+/- SD of 0.9 h). This is intermediate between the half-lives of its 2 components (approximately 45 minutes for IL-2 and 3 days for the chimeric m14.18 antibody), and comparable to that which was observed for the half-life of chimeric m14.18-IL-2 in mice. Following the clearance of hul4.18-IL-2 from the serum of these patients, neither the IL-2 nor hul4.18 antibody components could be detected.
The peak serum and area under the curve (AUC) during course 1 showed a significant dose-dependent increase (p<0.001).
Thirty-three patients were treated in this study. Table 1 lists clinical outcomes.
Two patients (6%) completed only the first 2 of 3 days for course 1. One of these patients (dose level 3) had a grade 3 hyperbilirubinemia on day 2 of treatment, and the other patient (dose level 6) had grade 3 hypoxia and hypotension requiring treatment to be held.
Both of these patients had progression of disease and did not receive a second course of therapy. Nineteen patients (58%) had stable disease following the first course of therapy and received a second course of therapy. Five patients (15% of all patients) required a 50% dose reduction for course 2 secondary to adverse events in course 1.
Seventeen patients (52% of all patients) completed course 2. One patient (dose level 4) declined to receive the final infusion during course 2, and one patient (dose level 6) had the final infusion during course 2 held due to hypotension. Eight patients (24% of all patients) had stable disease following the second course of treatment. The results indicate that hul4.18-IL-2 caused stabilization of disease progression in a surprisingly large number of patients.
Eight of the 33 patients maintained stable disease after 2 courses of therapy, and 4 of these 8 patients continue with no evidence of progressive disease (1 with stable disease and 3 with no evidence of disease) for 20-52 months since completing protocol therapy.
Five of the 33 patients entered the study with no measurable disease following surgical resection of recurrences or metastases. Two of these five patients had disease progression, while the remaining 3 patients continued with no evidence of disease (20-52 months). These findings are consistent with the hypothesis that clinical benefit from an immunotherapeutic intervention is most likely in a patient with a low tumor burden. One additional patient had an objective decrease in a lung nodule following two courses of therapy, but the overall disease response was scored as disease progression due to growth in a distant node. The node was resected following hul4.18-IL-2 therapy and the patient remained free from disease progression for over 3 years.
Clinical Outcomes Number of Patients Patients completing course 1 31 Stable disease following course 1 19 50% dose reduction for course 2 5 Patients completing course 2 17 Stable disease following course 2 8 Immune stimulation in vivo by hul4.18-IL-2 in a Phase I clinical trial.
Patients treated with hul4.18-IL-2 were also examined for indications of immune stimulation. A peripheral blood lymphopenia occurred on days 2-4, and this was followed by a rebound lymphocytosis on days 5-22. Both of these changes were dose-dependent (p<0.01 and p<0.05, respectively). The lymphocyte counts on days 5, 8, 15 and 22 were significantly greater than baseline for course 1. The baseline lymphocyte count for course 2 (day 29 of course 1) was increased over the baseline lymphocyte count for course 1, indicating that effects of the first course of treatment are still present on day 29. In addition, the lymphocyte counts during course 2 on days 5, 8 and 15 are greater than the corresponding values for days 5, 8, and 15 during course 1 for these 12 patients.
Lymphocyte cell surface phenotype showed an expansion of CD16+ and CD56+
lymphocytes (natural killer (NK) cell markers) following the first week of hul4.18-IL-2 therapy. This effect was still present on day 29 of course 1 (day 1, course 2). For patients 19-33 (receiving 4.8-7.5 mg/m2/day), lymphocyte cell surface phenotype was determined on days 15 and 22 in addition to days 1 and 8. This analysis demonstrated that the augmentation of CD56 and CD56/CD16 co-expressing cells remained significantly elevated (p<O.01) on days 8, 15 and 22.
As a measure of immune activation, C-reactive protein (CRP) levels for patients 13-33 and soluble IL-2 receptor (sIL-2R) levels for the 31 patients completing course 1, were obtained. A significant increase in mean CRP was present on treatment days 3-5 in both course 1 and course 2 compared to baseline for each course. This increase in CRP
returned to baseline levels by day 8 of each treatment course. The sIL-2R
level was significantly increased over baseline starting 24 hours after the hul4.18-IL-2 infusion during both course 1 and course 2, which persisted through day S. The increase in sIL-2R
was found to be dose dependent (p= 0.014). sIL-2R values for course 2 were increased compared to corresponding values in course 1 for days 1-5 for patients receiving the same dose in both courses (p < 0.05).
The LA-N-5 neuroblastoma cell line that expresses GD2 and binds hul4.18-IL-2 was used to evaluate IL-2 activated NK function and antibody dependent cellular cytotoxicity (ADCC) on peripheral blood mononuclear cells (PBMC) from 31 patients completing course 1. There was a significant increase in killing mediated by lymphocytes from day 8 'when compared with day 1 for these two assays. The 12 patients that received course 2 at the same dose as in course 1, showed ADCC results that were very similar to those obtained during course 1. The only parameter that was found to be different for course 2 from course 1 was increased killing in the presence of IL-2 on day 1, indicating that augmented killing in this assay remained elevated on day 29 (day 1, course 2).
Because the LA-N-5 target is relatively resistant to fresh NK cells, it is useful for measuring IL-2 augmented killing, and ADCC. However, the weak killing of LA-N-mediated by fresh PBMC in medium (without supplemental IL-2 in vitro) was not significantly greater on day 8 than on day 1.
For patients 19-33, standard NK assays were performed on days 1, 8, 15 and 22, using the NIA susceptible K562 target cell line. A significant increase in NK
lysis of K562 target cells, when tested either in medium or in the presence of IL-2, was observed on days 8 and 22 when compared with day 1. Serum samples from selected patients were also evaluated to determine functional IL-2 activity and functional anti-GD2 antibody.
The IL-2 responsive Tf-lb cell line demonstrated IL-2-induced proliferation with patient serum obtained following infusion of hul.4.18-IL-2. A progressive increase in proliferation was seen during the first 4 hours following the 4-hour infusion.
Values returned to baseline by 16 hours after this infusion, consistent with the serum half-life for hul4.18-IL-2 of approximately 4 hours. Serum samples from these time-points were also examined by flow cytometry for the presence of intact hul4.18-IL-2 immunocytokine (IC) that retains its IL-2 component and its anti-GD2 antibody activity.
hul4.18-IL-2 capable of binding to the M21 cell line (GD2 positive) was detectable in patient serum samples following an infusion of IC. The amount of IC able to bind to M21 progressively increased during the first 4 hours following the 4-hour infusion, and decreased after that, again consistent with the half-life of approximately 4 hours.
Finally, in vitro assays were performed with specimens from patients to determine whether administration of hul4.18-IL-2 results in conditions in vivo consistent with those needed to achieve ADCC. PBMCs from day 8 show augmented ADCC on GD2+ target cells when hul4.18-IL-2 is added to the cytotoxic assay. This same ADCC assay was performed with PBMC from day 8, however instead of adding hul4.18-IL-2 to the assay, serum from the patient, obtained before or after hul4.18-IL-2 administration, was added.
PBMC obtained from patients on day 8 of course 2 were able to mediate augmented killing of the LA-N-5 cell line in the presence of serum obtained following hul4.18-IL-2 administration, compared to that observed with serum obtained prior to infusion. Thus the hul4.18-IL-2 circulating in patients after IV administration is able to facilitate ADCC
with PBMCs activated in vivo by hul4.18-IL-2 from that same patient.
In summary, these results indicate that there were immunological changes associated with this hul4.18-IL-2 therapy including an increase in lymphocyte count, an increase in the percentage of CD16+ and CD56+ PBMC, an increase in NK lysis, and an increase in ADCC. Additional evidence for immune activation included an increase in serum levels of CRP and of sIL-2R. Laboratory analyses of serum and PBMC
showed that the hul4.18-IL-2 molecule circulating in patient serum following IV
administration retained its ability to activate IL-2 responsive cells through the IL-2 receptor and retained its ability to bind to GD2 positive tumor cells, and deliver IL-2 to their surface, as detected by flow cytometry. NK cells were activated in vivo based on their ability to mediate NK and ADCC function in vitro. Furthermore, the NK cells activated in vivo by the hul4.18-IL-2 administered to these patients were able to mediate ADCC
facilitated by the hul4.l8-IL-2 circulating in the serum of those same patients. Thus, conditions to achieve immune activation were achieved in all patients in this study.
SEQUENCE LISTING
<110> Gillies , Stephen D.
Lo, Kin-Ming <120> Immunocytokine Sequences and Uses Thereof <130> LEX-023 <150> US 60/433,945 <151> 2002-12-17 <160> 6 <170> Patentln version 3.1 <210> 1 <211> 113 <212> PRT
<213> Artificial Sequence <220>
<223> Humanized Immunoglobulin light chain variable region <400> 1 Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Arg Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gin Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile His Lys Val Ser Asn Arg Phe Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Ser Thr His Val Pro Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys <210> 2 <211> 113 <212> PRT
<213> Artificial Sequence <220>
<223> Humanized Immunoglobulin heavy chain variable region <400> 2 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Glu Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Ser Ser Phe Thr Gly Tyr Asn Met Asn Trp Val Arg Gln Asn Ile Gly Lys Ser Leu Glu Trp Ile Gly Ala Ile Asp Pro Tyr Tyr Gly Gly Thr Ser Tyr Asn Gln Lys Phe Lys Gly Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr Met His Leu Lys Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Val Ser Gly Met Glu Tyr Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser <210> 3 <211> 15 <212> PRT
<213> Artificial Sequence <220>
<223> Linker sequence <400> 3 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser <210> 4 <211> 10531 <212> DNA
<213> Artificial Sequence <220>
<223> Vector containing humanized Immunoglobulin light and heavy chain and IL-2 <400> 4 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 60 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 120 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 180 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 240 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 300 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 360 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 420 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 480 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 540 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta 600 cagaacccac tgcttaactg gcttatcgaa attaatacga ctcactatag ggagaccctc 660 tagaatgaag ttgcctgtta ggctgttggt gctgatgttc tggattcctg gtgaggagag 720 agggaagtga gggaggagaa tggacaggga gcaggagcac tgaatcccat tgctcattcc 780 atgtatctgg catgggtgag aagatgggtc ttatcctcca gcatggggcc tctggggtga 840 atacttgtta gagggaggtt ccagatggga acatgtgcta taatgaagat tatgaaatgg 900 atgcctggga tggtctaagt aatgccttag aagtgactag acacttgcaa ttcacttttt 960 ttggtaagaa gagattttta ggctataaaa aaatgttatg taaaaataaa cgatcacagt 1020 tgaaataaaa aaaaaatata aggatgttca tgaattttgt gtataactat gtatttctct 1080 ctcattgttt cagcttcctt aagcgacgtg gtgatgaccc agacccccct gtccctgccc 1140 gtgacccccg gcgagcccgc ctccatctcc tgcagatcta gtcagagtct tgtacaccgt 1200 aatggaaaca cctatttaca ttggtacctg cagaagccag gccagtctcc aaagctcctg 1260 attcacaaag tttccaaccg attttctggg gtcccagaca ggttcagtgg cagtggatca 1320 gggacagatt tcacactcaa gatcagcaga gtggaggctg aggatctggg agtttatttc 1380 tgttctcaaa gtacacatgt tcctccgctc acgttcggtg ctgggaccaa gctggagctg 1440 aaacgtatta gtgtgtcagg gtttcacaag agggactaaa gacatgtcag ctatgtgtga 1500 ctaatggtaa tgtcactaag ctgcgggatc ccgcaattct aaactctgag ggggtcggat 1560 gacgtggcca ttctttgcct aaagcattga gtttactgca aggtcagaaa agcatgcaaa 1620 gccctcagaa tggctgcaaa gagctccaac aaaacaattt agaactttat taaggaatag 1680 ggggaagcta ggaagaaact caaaacatca agattttaaa tacgcttctt ggtctccttg 1740 ctataattat ctgggataag catgctgttt tctgtctgtc cctaacatgc cctgtgatta 1800 tccgcaaaca acacacccaa gggcagaact ttgttactta aacaccatcc tgtttgcttc 1860 tttcctcagg aactgtggct gcaccatctg tcttcatctt cccgccatct gatgagcagt 1920 tgaaatctgg aactgcctct gttgtgtgcc tgctgaataa cttctatccc agagaggcca 1980 aagtacagtg gaaggtggat aacgccctcc aatcgggtaa ctcccaggag agtgtcacag 2040 agcaggacag caaggacagc acctacagcc tcagcagcac cctgacgctg agcaaagcag 2100 actacgagaa acacaaagtc tacgcctgcg aagtcaccca tcagggcctg agctcgcccg 2160 tcacaaagag cttcaacagg ggagagtgtt agagggagaa gtgcccccac ctgctcctca 2220 gttccagcct gaccccctcc catcctttgg cctctgaccc tttttccaca ggggacctac 2280 ccctattgcg gtcctccagc tcatctttca cctcaccccc ctcctcctcc ttggctttaa 2340 ttatgctaat gttggaggag aatgaataaa taaagtgaat ctttgcacct gtggtttctc 2400 tctttcctca atttaataat tattatctgt tgtttaccaa ctactcaatt tctcttataa 2460 gggactaaat atgtagtcat cctaaggcgc ataaccattt ataaaaatca tccttcattc 2520 tattttaccc tatcatcctc tgcaagacag tcctccctca aacccacaag ccttctgtcc 2580 tcacagtccc ctgggccatg gtaggagaga cttgcttcct tgttttcccc tcctcagcaa 2640 gccctcatag tcctttttaa gggtgacagg tcttacggtc atatatcctt tgattcaatt 2700 ccctgggaat caaccaaggc aaatttttca aaagaagaaa cctgctataa agagaatcat 2760 tcattgcaac atgatataaa ataacaacac aataaaagca attaaataaa caaacaatag 2820 ggaaatgttt aagttcatca tggtacttag acttaatgga atgtcatgcc ttatttacat 2880 ttttaaacag gtactgaggg actcctgtct gccaagggcc gtattgagta ctttccacaa 2940 cctaatttaa tccacactat actgtgagat taaaaacatt cattaaaatg ttgcaaaggt 3000 tctataaagc tgagagacaa atatattcta taactcagca atcccacttc tagggtcgat 3060 cgacgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt 3120 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga 3180 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca 3240 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca 3300 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg 3360 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 3420 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 3480 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 3540 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 3600 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta 3660 actacagaac ccactgctta actggcttat cgaaattaat acgactcact atagggagac 3720 ccaagctcct cgaggctaga atgaagttgc ctgttaggct gttggtgctg atgttctgga 3780 ttcctggtga ggagagaggg aagtgaggga ggagaatgga cagggagcag gagcactgaa 3840 tcccattgct cattccatgt atctggcatg ggtgagaaga tgggtcttat cctccagcat 3900 ggggcctctg gggtgaatac ttgttagagg gaggttccag atgggaacat gtgctataat 3960 gaagattatg aaatggatgc ctgggatggt ctaagtaatg ccttagaagt gactagacac 4020 ttgcaattca ctttttttgg taagaagaga tttttaggct ataaaaaaat gttatgtaaa 4080 aataaacgat cacagttgaa ataaaaaaaa aatataagga tgttcatgaa ttttgtgtat 4140 aactatgtat ttctctctca ttgtttcagc ttccttaagc gaggtgcagc tggtgcagtc 4200 cggcgccgag gtggagaagc ccggcgcctc cgtgaagatc tcctgcaagg,cctccggctc 4260 ctccttcacc ggctacaaca tgaactgggt gcgccagaac atcggcaagt ccctggagtg 4320 gatcggcgcc atcgacccct actacggcgg cacctcctac aaccagaagt tcaagggccg 4380 cgccaccctg accgtggaca agtccacctc caccgcctac atgcacctga agtccctgcg 4440 ctccgaggac accgccgtgt actactgcgt gtccggcatg gagtactggg gccagggcac 4500 ctccgtgacc gtgtcctccg gtaagctttt ctggggcagg ccaggcctga ccttggcttt 4560 ggggcaggga gggggctaag gtgaggcagg tggcgccagc caggtgcaca cccaatgccc 4620 atgagcccag acactggacg ctgaacctcg cggacagtta agaacccagg ggcctctgcg 4680 ccctgggccc agctctgtcc cacaccgcgg tcacatggca ccacctctct tgcagcctcc 4740 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca 4800 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 4860 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 4920 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 4980 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agagagttgg tgagaggcaa 5040 gcacagggag ggagggtgtc tgctggaagc caggctcagc gctcctgcct ggacgcatcc 5100 cggctatgca gtcccagtcc agggcagcaa ggcaggcccc gtctgcctct tcacccggag 5160 gcctctgccc gccccactca tgctcaggga gagggtcttc tggctttttc cccaggctct 5220 gggcaggcac aggctaggtg cccctaaccc aggccctgca cacaaagggg caggtgctgg 5280 gctcagacct gccaagagcc atatccggga ggaccctgcc cctgacctaa gcccacccca 5340 aaggccaaac tctccactcc ctcagctcgg acaccttctc tcctcccaga ttccagtaac 5400 tcccaatctt ctctctgcag agcccaaatc ttgtgacaaa actcacacat gcccaccgtg 5460 cccaggtaag ccagcccagg cctcgccctc cagctcaagg cgggacaggt gccctagagt 5520 agcctgcatc cagggacagg ccccagccgg gtgctgacac gtccacctcc atctcttcct 5580 cagcacctga actcctgggg ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca 5640 ccctcatgat ctcccggacc cctgaggtca catgcgtggt ggtggacgtg agccacgaag 5700 accctgaggt caagttcaac tggtacgtgg acggcgtgga ggtgcataat gccaagacaa 5760 agccgcggga ggagcagtac aacagcacgt accgtgtggt cagcgtcctc accgtcctgc 5820 accaggactg gctgaatggc aaggagtaca agtgcaaggt ctccaacaaa gccctcccag 5880 cccccatcga gaaaaccatc tccaaagcca aaggtgggac ccgtggggtg cgagggccac 5940 atggacagag gccggctcgg cccaccctct gccctgagag tgaccgctgt accaacctct 6000 gtccctacag ggcagccccg agaaccacag gtgtacaccc tgcccccatc acgggaggag 6060 atgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc 6120 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 6180 ctggactccg acggctcctt cttcctctat agcaagctca ccgtggacaa gagcaggtgg 6240 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 6300 cagaagagcc tctccctgtc cccgggtaaa gccccaactt caagttctac aaagaaaaca 6360 cagctgcaac tggagcatct cctgctggat ctccagatga ttctgaatgg aattaacaac 6420 tacaagaatc ccaaactcac caggatgctc acattcaagt tctacatgcc caagaaggcc 6480 acagagctca aacatctcca gtgtctagag gaggaactca aacctctgga ggaagtgcta 6540 aacctcgctc agagcaaaaa cttccactta agacctaggg acttaatcag caatatcaac 6600 gtaatagttc tggaactaaa gggatccgaa acaacattca tgtgtgaata tgctgatgag 6660 acagcaacca ttgtagaatt tctgaacaga tggattacct tttgtcaaag catcatctca 6720 acactaactt gataattaag tgctcgaggg atccagacat gataagatac attgatgagt 6780 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg 6840 ctattgcttt atttgtaacc attagaagct gcaataaaca agttaacaac aacaattgca 6900 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc 6960 tctacaaatg tggtatggct gattatgatc ctgcctcgcg cgtttcggtg atgacggtga 7020 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg 7080 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat 7140 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag 7200 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa 7260 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 7320 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 7380 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 7440 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 7500 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 7560 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 7620 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg 7680 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 7740 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 7800 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 7860 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct 7920 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 7980 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 8040 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 8100 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 8160 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 8220 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 8280 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 8340 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 8400 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 8460 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 8520 gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 8580 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 8640 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 8700 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 8760 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 8820 tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 8880 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 8940 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 9000 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 9060 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 9120 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 9180 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta 9240 acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaattccg atccagacat 9300 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt 9360 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attagaagct gcaataaaca 9420 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt 9480 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc taaagccagc 9540 aaaagtccca tggtcttata aaaatgcata gctttcggag gggagcagag aacttgaaag 9600 catcttcctg ttagtctttc ttctcgtaga ccttaaattc atacttgatt cctttttcct 9660 cctggacctc agagaggacg cctgggtatt ctgggagaag tttatatttc cccaaatcaa 9720 tttctgggaa aaacgtgtca ctttcaaatt cctgcatgat ccttgtcaca aagagtctga 9780 ggtggcctgg ttgattcatg gcttcctggt aaacagaact gcctccgact atccaaacca 9840 tgtctacttt acttgccaat tccggttgtt caataagtct taaggcatca tccaaacttt 9900 tggcaagaaa atgagctcct cgtggtggtt ctttgagttc tctactgaga actatattaa 9960 ttctgtcctt taaaggtcga ttcttctcag gaatggagaa ccaggttttc ctacccataa 10020 tcaccagatt ctgtttacct tccactgaag aggttgtggt cattctttgg aagtacttga 10080 actcgttcct gagcggaggc cagggtcggt ctccgttctt gccaatcccc atattttggg 10140 acacggcgac gatgcagttc aatggtcgaa ccatgagggc accaagctag ctttttgcaa 10200 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg 10260 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa 10320 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa 10380 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg 10440 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac 10500 tttccacacc ctaactgaca cacattccac a 10531 <210> 5 <211> 220 <212> PRT
<213> Artificial Sequence <220>
<223> Humanized Immunoglobulin light chain <400> 5 Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Arg Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile His Lys Val Ser Asn Arg Phe Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Ser Thr His Val Pro Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys <210> 6 <211> 575 <212> PRT
<213> Artificial Sequence <220>
<223> Humanized Immunoglobulin heavy chain and IL-2 <400> 6 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Glu Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Ser Ser Phe Thr Gly Tyr Asn Met Asn Trp Val Arg Gln Asn Ile Gly Lys Ser Leu Glu Trp Ile Gly Ala Ile Asp Pro Tyr Tyr Gly Gly Thr Ser Tyr Asn Gln Lys Phe Lys Gly Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr Met His Leu Lys Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Val Ser Gly Met Glu Tyr Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile Ile Ser Thr Leu Thr
Claims (7)
1. A humanized antibody-IL2 fusion protein designated as hu14.18-IL2 that specifically binds GD2 and stimulates immune function comprising the light chain of SEQ ID NO. 5 and the heavy chain of SEQ ID NO. 6.
2. A vector comprising the nucleotide sequence of SEQ ID NO. 4 containing the nucleic acid sequences that code for the fusion protein of claim 1.
3. Pharmaceutical composition comprising the fusion protein of claim 1 and a pharmaceutical carrier or excipient.
4. Use of the fusion protein of claim 1 for the manufacture of a medicament for stabilizing disease progression in GD2 positive cancer patients.
5. Use of the fusion protein of claim 1 for the manufacture of a medicament for increasing ADCC and NK-lysis activity in GD2 positive cancer patients.
6. A pharmaceutical composition as claimed in claim 3, for use in stabilizing disease progression in GD2 positive cancer patients.
7. A pharmaceutical composition as claimed in claim 3, for use in increasing ADCC and NK-lysis activity in GD2 positive cancer patients.
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US43394502P | 2002-12-17 | 2002-12-17 | |
US60/433,945 | 2002-12-17 | ||
PCT/EP2003/014295 WO2004055056A1 (en) | 2002-12-17 | 2003-12-16 | Humanized antibody (h14.18) of the mouse 14.18 antibody binding to gd2 and its fusion with il-2 |
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CA2510180A Expired - Lifetime CA2510180C (en) | 2002-12-17 | 2003-12-16 | Humanized antibody (h14.18) of the mouse 14.18 antibody binding to gd2 and its fusion with il-2 |
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Families Citing this family (63)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
ATE267215T1 (en) * | 1997-12-08 | 2004-06-15 | Lexigen Pharm Corp | HETERODIMARY FUSION PROTEINS FOR USE FOR TARGETED IMMUNTHERAPY AND GENERAL IMMUNE EXCITATION |
US20030105294A1 (en) * | 1998-02-25 | 2003-06-05 | Stephen Gillies | Enhancing the circulating half life of antibody-based fusion proteins |
SK782002A3 (en) | 1999-07-21 | 2003-08-05 | Lexigen Pharm Corp | FC fusion proteins for enhancing the immunogenicity of protein and peptide antigens |
US20050202538A1 (en) * | 1999-11-12 | 2005-09-15 | Merck Patent Gmbh | Fc-erythropoietin fusion protein with improved pharmacokinetics |
CA2399832C (en) | 2000-02-11 | 2011-09-20 | Stephen D. Gillies | Enhancing the circulating half-life of antibody-based fusion proteins |
ES2288967T3 (en) * | 2000-06-29 | 2008-02-01 | Merck Patent Gmbh | REINFORCEMENT OF IMMUNE ANSWERS MEDIATED BY THE ANTIBODY-CYTOKIN FUSION PROTEIN THROUGH THE TREATMENT COMBINED BY AGENTS THAT IMPROVE THE INCORPORATION OF IMMUNOCITOQUINE. |
CA2440221C (en) | 2001-03-07 | 2013-02-05 | Merck Patent Gesellschaft Mit Beschraenkter Haftung | Expression technology for proteins containing a hybrid isotype antibody moiety |
WO2002079415A2 (en) | 2001-03-30 | 2002-10-10 | Lexigen Pharmaceuticals Corp. | Reducing the immunogenicity of fusion proteins |
BR0209177A (en) | 2001-05-03 | 2004-10-05 | Merck Patent Gmbh | Recombinant tumor specific antibody and use |
ES2381025T3 (en) | 2001-12-04 | 2012-05-22 | Merck Patent Gmbh | Immunocytokines with modulated selectivity |
US10001475B2 (en) | 2002-06-20 | 2018-06-19 | The Regents Of The University Of California | Light harvesting multichromophore compositions and methods of using the same |
US9371559B2 (en) | 2002-06-20 | 2016-06-21 | The Regents Of The University Of California | Compositions for detection and analysis of polynucleotides using light harvesting multichromophores |
ES2346205T3 (en) * | 2002-12-17 | 2010-10-13 | Merck Patent Gmbh | HUMANIZED ANTIBODY (H14.18) OF ANTIBODY 14.18 OF MOUSE THAT LINKS TO GD2 AND ITS FUSION WITH IL-2. |
CN1771335A (en) * | 2003-02-13 | 2006-05-10 | 加州大学评议会 | Methods and compositions for detection and analysis of polynucleotide-binding protein interactions using light harvesting multichromophores |
US20050069521A1 (en) * | 2003-08-28 | 2005-03-31 | Emd Lexigen Research Center Corp. | Enhancing the circulating half-life of interleukin-2 proteins |
US7642341B2 (en) | 2003-12-18 | 2010-01-05 | Merck Serono S.A. | Angiogenesis inhibiting molecules, their selection, production and their use in the treatment of cancer |
EP1533617A1 (en) | 2003-11-19 | 2005-05-25 | RMF Dictagene S.A. | Angiogenesis inhibiting molecules, their selection, production and their use in the treatment and diagnosis of cancer |
WO2005063820A2 (en) | 2003-12-30 | 2005-07-14 | Merck Patent Gmbh | Il-7 fusion proteins |
KR20060124656A (en) * | 2003-12-31 | 2006-12-05 | 메르크 파텐트 게엠베하 | Fc-erythropoietin fusion protein with improved pharmacokinetics |
CA2553883C (en) | 2004-01-22 | 2013-04-02 | Merck Patent Gesellschaft Mit Beschraenkter Haftung | Anti-cancer antibodies with reduced complement fixation |
US7670595B2 (en) * | 2004-06-28 | 2010-03-02 | Merck Patent Gmbh | Fc-interferon-beta fusion proteins |
DE602005020837D1 (en) * | 2004-12-09 | 2010-06-02 | Merck Patent Gmbh | IL-7 VARIANTS WITH REDUCED IMMUNOGENITY |
US20070104689A1 (en) * | 2005-09-27 | 2007-05-10 | Merck Patent Gmbh | Compositions and methods for treating tumors presenting survivin antigens |
CA2635618C (en) * | 2005-12-30 | 2015-10-06 | Merck Patent Gesellschaft Mit Beschraekter Haftung | Interleukin-12p40 variants with improved stability |
PT2270050E (en) | 2005-12-30 | 2013-09-12 | Merck Patent Gmbh | Anti-cd19 antibodies with reduced immunogenicity |
PL1999152T3 (en) * | 2006-03-27 | 2013-05-31 | Medimmune Ltd | Binding member for gm-csf receptor |
US7787618B2 (en) | 2006-03-29 | 2010-08-31 | Nokia Corporation | Portable electronic device |
EP2038417A2 (en) * | 2006-07-06 | 2009-03-25 | Merck Patent GmbH | Compositions and methods for enhancing the efficacy of il-2 mediated immune responses |
JP2010504955A (en) | 2006-09-28 | 2010-02-18 | メルク セローノ ソシエテ アノニム | Bonding molecule adhesion molecule C (JAM-C) binding compound and method of use thereof |
WO2008045380A2 (en) * | 2006-10-04 | 2008-04-17 | Codon Devices, Inc. | Nucleic acid libraries and their design and assembly |
FR2906808B1 (en) * | 2006-10-10 | 2012-10-05 | Univ Nantes | USE OF MONOCLONAL ANTIBODIES SPECIFIC TO THE O-ACETYLATED FORMS OF GANGLIOSIDE GD2 IN THE TREATMENT OF CERTAIN CANCERS |
WO2009152901A1 (en) * | 2008-05-30 | 2009-12-23 | Merck Patent Gmbh, | Methods for treatment of malignancies |
WO2009149218A2 (en) * | 2008-06-03 | 2009-12-10 | Codon Devices, Inc. | Novel proteins and methods of designing and using same |
US8278065B2 (en) * | 2008-06-30 | 2012-10-02 | Morphotek, Inc. | Polynucleotides encoding anti-GD2 antibodies |
WO2010117448A2 (en) * | 2009-04-05 | 2010-10-14 | Provenance Biopharmaceuticals Corp. | Chimeric immunocytokines and methods of use thereof |
AU2010238858A1 (en) | 2009-04-22 | 2011-12-08 | Merck Patent Gmbh | Antibody fusion proteins with modified FcRn binding sites |
JP5638628B2 (en) * | 2010-01-19 | 2014-12-10 | ハンミ サイエンス カンパニー リミテッドHanmi Scienceco.,Ltd. | Long-acting erythropoietin conjugate solution |
PT2560658T (en) * | 2010-04-21 | 2017-05-26 | Ventirx Pharmaceuticals Inc | Enhancing antibody-dependent cellular cytotoxicity |
WO2011141926A2 (en) * | 2010-05-10 | 2011-11-17 | Intas Biopharmaceuticals Limited | Liquid formulation of polypeptides containing an fc domain of an immunoglobulin |
ES2961381T3 (en) | 2010-06-19 | 2024-03-11 | Memorial Sloan Kettering Cancer Center | Anti-GD2 antibodies |
US20120065092A1 (en) * | 2010-09-14 | 2012-03-15 | Wai Hobert | Fusion analyte cytometric bead assay, and systems and kits for performing the same |
EP3971206A1 (en) * | 2011-02-10 | 2022-03-23 | Roche Glycart AG | Mutant interleukin-2 polypeptides |
EA201892619A1 (en) | 2011-04-29 | 2019-04-30 | Роше Гликарт Аг | IMMUNOCONJUGATES CONTAINING INTERLEUKIN-2 MUTANT POLYPETIPS |
US11492383B2 (en) | 2011-06-24 | 2022-11-08 | Stephen D. Gillies | Light chain immunoglobulin fusion proteins and methods of use thereof |
US9777068B2 (en) | 2012-06-18 | 2017-10-03 | Apeiron Biologics Ag | Method for treating a GD2 positive cancer |
WO2013189516A1 (en) * | 2012-06-18 | 2013-12-27 | Apeiron Biologics Ag | Method for treating a gd2 positive cancer |
JOP20140087B1 (en) | 2013-03-13 | 2021-08-17 | Amgen Inc | Proteins specific for baff and b7rp1 and uses thereof |
US9458246B2 (en) | 2013-03-13 | 2016-10-04 | Amgen Inc. | Proteins specific for BAFF and B7RP1 |
CA2903576C (en) * | 2013-03-15 | 2021-06-08 | Nai-Kong V. Cheung | High affinity anti-gd2 antibodies |
US20160040212A1 (en) * | 2013-03-15 | 2016-02-11 | The Broad Institute, Inc. | Methods for the Detection of DNA-RNA Proximity in Vivo |
NZ720273A (en) | 2013-11-21 | 2022-02-25 | Apeiron Biologics Ag | Preparations and methods for treating a gd2 positive cancer |
US9840566B2 (en) | 2013-11-21 | 2017-12-12 | Apeiron Biologics Ag | Preparations and methods for treating a GD2 positive cancer |
CN105873614B (en) * | 2013-12-16 | 2020-10-30 | 基因泰克公司 | Peptidomimetic compounds and antibody-drug conjugates thereof |
GB201403775D0 (en) | 2014-03-04 | 2014-04-16 | Kymab Ltd | Antibodies, uses & methods |
GB201403972D0 (en) | 2014-03-06 | 2014-04-23 | Ucl Business Plc | Chimeric antigen receptor |
WO2017055385A1 (en) * | 2015-10-02 | 2017-04-06 | F. Hoffmann-La Roche Ag | Anti-cd3xgd2 bispecific t cell activating antigen binding molecules |
US9567399B1 (en) | 2016-06-20 | 2017-02-14 | Kymab Limited | Antibodies and immunocytokines |
JP7438756B2 (en) * | 2016-07-25 | 2024-02-27 | ウイスコンシン アラムナイ リサーチ ファウンデーシヨン | Targeted radiotherapeutic chelates for in situ immunomodulatory cancer vaccination |
US11779604B2 (en) | 2016-11-03 | 2023-10-10 | Kymab Limited | Antibodies, combinations comprising antibodies, biomarkers, uses and methods |
US11129906B1 (en) | 2016-12-07 | 2021-09-28 | David Gordon Bermudes | Chimeric protein toxins for expression by therapeutic bacteria |
CN108948211B (en) * | 2018-07-24 | 2021-08-20 | 北京美康基免生物科技有限公司 | Chimeric antigen receptor based on targeting GD2 and application thereof |
RU2708136C1 (en) * | 2019-04-15 | 2019-12-04 | Общество с ограниченной ответственностью "Реал Таргет" | Novel protein structures for diagnosing and therapy of gd2-positive diseases |
WO2024040194A1 (en) | 2022-08-17 | 2024-02-22 | Capstan Therapeutics, Inc. | Conditioning for in vivo immune cell engineering |
Family Cites Families (171)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4196265A (en) * | 1977-06-15 | 1980-04-01 | The Wistar Institute | Method of producing antibodies |
US4522811A (en) * | 1982-07-08 | 1985-06-11 | Syntex (U.S.A.) Inc. | Serial injection of muramyldipeptides and liposomes enhances the anti-infective activity of muramyldipeptides |
US4469797A (en) * | 1982-09-23 | 1984-09-04 | Miles Laboratories, Inc. | Digoxigenin immunogens, antibodies, labeled conjugates, and related derivatives |
US4737462A (en) * | 1982-10-19 | 1988-04-12 | Cetus Corporation | Structural genes, plasmids and transformed cells for producing cysteine depleted muteins of interferon-β |
US4966843A (en) * | 1982-11-01 | 1990-10-30 | Cetus Corporation | Expression of interferon genes in Chinese hamster ovary cells |
US4816567A (en) * | 1983-04-08 | 1989-03-28 | Genentech, Inc. | Recombinant immunoglobin preparations |
NZ210501A (en) * | 1983-12-13 | 1991-08-27 | Kirin Amgen Inc | Erythropoietin produced by procaryotic or eucaryotic expression of an exogenous dna sequence |
KR850004274A (en) * | 1983-12-13 | 1985-07-11 | 원본미기재 | Method for preparing erythropoietin |
US4703008A (en) * | 1983-12-13 | 1987-10-27 | Kiren-Amgen, Inc. | DNA sequences encoding erythropoietin |
US5082658A (en) * | 1984-01-16 | 1992-01-21 | Genentech, Inc. | Gamma interferon-interleukin-2 synergism |
EP0158198A1 (en) | 1984-03-29 | 1985-10-16 | Takeda Chemical Industries, Ltd. | DNA and use thereof |
US5807715A (en) * | 1984-08-27 | 1998-09-15 | The Board Of Trustees Of The Leland Stanford Junior University | Methods and transformed mammalian lymphocyte cells for producing functional antigen-binding protein including chimeric immunoglobulin |
US4667016A (en) * | 1985-06-20 | 1987-05-19 | Kirin-Amgen, Inc. | Erythropoietin purification |
US4690915A (en) | 1985-08-08 | 1987-09-01 | The United States Of America As Represented By The Department Of Health And Human Services | Adoptive immunotherapy as a treatment modality in humans |
US5679543A (en) * | 1985-08-29 | 1997-10-21 | Genencor International, Inc. | DNA sequences, vectors and fusion polypeptides to increase secretion of desired polypeptides from filamentous fungi |
US5643565A (en) * | 1985-09-20 | 1997-07-01 | Chiron Corporation | Human IL-2 as a vaccine adjuvant |
US4676980A (en) * | 1985-09-23 | 1987-06-30 | The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services | Target specific cross-linked heteroantibodies |
US4935233A (en) * | 1985-12-02 | 1990-06-19 | G. D. Searle And Company | Covalently linked polypeptide cell modulators |
US5359035A (en) | 1985-12-21 | 1994-10-25 | Hoechst Aktiengesellschaft | Bifunctional proteins including interleukin-2 (IL-2) and granuloctyte macrophage colony stimulating factor (GM-CSF) |
DE3712985A1 (en) | 1987-04-16 | 1988-11-03 | Hoechst Ag | BIFUNCTIONAL PROTEINS |
EP0237019A3 (en) | 1986-03-14 | 1988-03-09 | Toray Industries, Inc. | Interferon conjugate and production thereof using recombinant gene |
US5225539A (en) * | 1986-03-27 | 1993-07-06 | Medical Research Council | Recombinant altered antibodies and methods of making altered antibodies |
GB8607679D0 (en) | 1986-03-27 | 1986-04-30 | Winter G P | Recombinant dna product |
DK173067B1 (en) * | 1986-06-27 | 1999-12-13 | Univ Washington | Human erythropoietin gene, method of expression thereof in transfected cell lines, the transfected cell lines |
US4894227A (en) | 1986-08-01 | 1990-01-16 | Cetus Corporation | Composition of immunotoxins with interleukin-2 |
US4946778A (en) * | 1987-09-21 | 1990-08-07 | Genex Corporation | Single polypeptide chain binding molecules |
US4844893A (en) * | 1986-10-07 | 1989-07-04 | Scripps Clinic And Research Foundation | EX vivo effector cell activation for target cell killing |
US5508031A (en) * | 1986-11-21 | 1996-04-16 | Cetus Oncology Corporation | Method for treating biological damage using a free-radial scavenger and interleukin-2 |
US4732683A (en) * | 1986-12-02 | 1988-03-22 | Biospectrum, Inc. | Purification method for alpha interferon |
US5019368A (en) * | 1989-02-23 | 1991-05-28 | Cancer Biologics, Inc. | Detection of necrotic malignant tissue and associated therapy |
EP0307434B2 (en) * | 1987-03-18 | 1998-07-29 | Scotgen Biopharmaceuticals, Inc. | Altered antibodies |
US5258498A (en) * | 1987-05-21 | 1993-11-02 | Creative Biomolecules, Inc. | Polypeptide linkers for production of biosynthetic proteins |
DE3856559T2 (en) | 1987-05-21 | 2004-04-29 | Micromet Ag | Multifunctional proteins with predetermined objectives |
US5091513A (en) * | 1987-05-21 | 1992-02-25 | Creative Biomolecules, Inc. | Biosynthetic antibody binding sites |
ATE122238T1 (en) | 1987-06-10 | 1995-05-15 | Dana Farber Cancer Inst Inc | BIFUNCTIONAL ANTIBODY CONSTRUCTIONS AND METHOD FOR SELECTIVE KILLING OF CELLS. |
US5064646A (en) | 1988-08-02 | 1991-11-12 | The University Of Maryland | Novel infectious bursal disease virus |
ES2054753T3 (en) | 1987-09-02 | 1994-08-16 | Ciba Geigy Ag | CONJUGATES OF CYTOKINES WITH IMMUNOGLOBULINS. |
EP0308936B1 (en) | 1987-09-23 | 1994-07-06 | Bristol-Myers Squibb Company | Antibody heteroconjugates for the killing of HIV-infected cells |
NZ226414A (en) | 1987-10-02 | 1992-07-28 | Genentech Inc | Cd4 peptide adhesion variants and their preparation and use |
AU2635088A (en) | 1987-12-04 | 1989-06-08 | Du Pont Merck Pharmaceutical Company, The | Immobilized interleukin 2 and interleukin 2 containing a carboxyl-terminal extension |
WO1989006692A1 (en) * | 1988-01-12 | 1989-07-27 | Genentech, Inc. | Method of treating tumor cells by inhibiting growth factor receptor function |
CA1341588C (en) | 1988-01-26 | 2009-01-06 | Michel Revel | Human ifn-beta2/i1-6, its purification and use |
US5120525A (en) * | 1988-03-29 | 1992-06-09 | Immunomedics, Inc. | Radiolabeled antibody cytotoxic therapy of cancer |
US4975369A (en) | 1988-04-21 | 1990-12-04 | Eli Lilly And Company | Recombinant and chimeric KS1/4 antibodies directed against a human adenocarcinoma antigen |
IE62463B1 (en) | 1988-07-07 | 1995-02-08 | Res Dev Foundation | Immunoconjugates for cancer diagnosis and therapy |
US5601819A (en) * | 1988-08-11 | 1997-02-11 | The General Hospital Corporation | Bispecific antibodies for selective immune regulation and for selective immune cell binding |
US5457038A (en) * | 1988-11-10 | 1995-10-10 | Genetics Institute, Inc. | Natural killer stimulatory factor |
US5242824A (en) | 1988-12-22 | 1993-09-07 | Oncogen | Monoclonal antibody to human carcinomas |
US5530101A (en) | 1988-12-28 | 1996-06-25 | Protein Design Labs, Inc. | Humanized immunoglobulins |
US5225538A (en) * | 1989-02-23 | 1993-07-06 | Genentech, Inc. | Lymphocyte homing receptor/immunoglobulin fusion proteins |
US5116964A (en) * | 1989-02-23 | 1992-05-26 | Genentech, Inc. | Hybrid immunoglobulins |
US5703055A (en) | 1989-03-21 | 1997-12-30 | Wisconsin Alumni Research Foundation | Generation of antibodies through lipid mediated DNA delivery |
US5166322A (en) | 1989-04-21 | 1992-11-24 | Genetics Institute | Cysteine added variants of interleukin-3 and chemical modifications thereof |
ZA902949B (en) | 1989-05-05 | 1992-02-26 | Res Dev Foundation | A novel antibody delivery system for biological response modifiers |
US6291158B1 (en) * | 1989-05-16 | 2001-09-18 | Scripps Research Institute | Method for tapping the immunological repertoire |
US5399346A (en) * | 1989-06-14 | 1995-03-21 | The United States Of America As Represented By The Department Of Health And Human Services | Gene therapy |
ATE123065T1 (en) * | 1989-07-07 | 1995-06-15 | Takeda Chemical Industries Ltd | PROTEINS AND THEIR PRODUCTION. |
US5073627A (en) | 1989-08-22 | 1991-12-17 | Immunex Corporation | Fusion proteins comprising GM-CSF and IL-3 |
US5196320A (en) * | 1989-09-20 | 1993-03-23 | Abbott Biotech, Inc. | Method of producing engineered binding proteins |
WO1991005867A1 (en) | 1989-10-13 | 1991-05-02 | Amgen Inc. | Erythropoietin isoforms |
US5856298A (en) * | 1989-10-13 | 1999-01-05 | Amgen Inc. | Erythropoietin isoforms |
US5859205A (en) | 1989-12-21 | 1999-01-12 | Celltech Limited | Humanised antibodies |
ATE163675T1 (en) | 1989-12-22 | 1998-03-15 | Hoffmann La Roche | CYTOTOXIC LYMPHOCYTE MATURATION FACTOR |
US5314995A (en) * | 1990-01-22 | 1994-05-24 | Oncogen | Therapeutic interleukin-2-antibody based fusion proteins |
US5349053A (en) * | 1990-06-01 | 1994-09-20 | Protein Design Labs, Inc. | Chimeric ligand/immunoglobulin molecules and their uses |
US7253264B1 (en) | 1990-06-28 | 2007-08-07 | Sanofi-Arentideutschland GmbH | Immunoglobulin fusion proteins, their production and use |
ES2088499T3 (en) * | 1990-09-18 | 1996-08-16 | Akzo Nobel Nv | COPOLYMERIZATION AND OPTICAL COPOLYMER PROCEDURE PRODUCED FROM THE SAME. |
US5650150A (en) | 1990-11-09 | 1997-07-22 | Gillies; Stephen D. | Recombinant antibody cytokine fusion proteins |
FR2670039B1 (en) * | 1990-11-29 | 1993-12-24 | Commissariat A Energie Atomique | METHOD AND DEVICE FOR RECONSTRUCTING THREE-DIMENSIONAL IMAGES OF AN OBJECT USING TWO CIRCULAR ACQUISITION PATHWAYS. |
US5709859A (en) * | 1991-01-24 | 1998-01-20 | Bristol-Myers Squibb Company | Mixed specificity fusion proteins |
US6072039A (en) | 1991-04-19 | 2000-06-06 | Rohm And Haas Company | Hybrid polypeptide comparing a biotinylated avidin binding polypeptide fused to a polypeptide of interest |
DE69233482T2 (en) | 1991-05-17 | 2006-01-12 | Merck & Co., Inc. | Method for reducing the immunogenicity of antibody variable domains |
US6797492B2 (en) * | 1991-05-17 | 2004-09-28 | Merck & Co., Inc. | Method for reducing the immunogenicity of antibody variable domains |
US5199942A (en) * | 1991-06-07 | 1993-04-06 | Immunex Corporation | Method for improving autologous transplantation |
WO1994004679A1 (en) | 1991-06-14 | 1994-03-03 | Genentech, Inc. | Method for making humanized antibodies |
DE69233254T2 (en) | 1991-06-14 | 2004-09-16 | Genentech, Inc., South San Francisco | Humanized Heregulin antibody |
ATE173763T1 (en) | 1991-08-30 | 1998-12-15 | Hutchinson Fred Cancer Res | HYBRID CYTOKINE |
US20020037558A1 (en) * | 1991-10-23 | 2002-03-28 | Kin-Ming Lo | E.coli produced immunoglobulin constructs |
EP0760493B1 (en) * | 1991-11-25 | 1999-01-20 | Sharp Kabushiki Kaisha | Device for further processing after copying |
US6627615B1 (en) * | 1991-12-17 | 2003-09-30 | The Regents Of The University Of California | Methods and compositions for in vivo gene therapy |
CZ291039B6 (en) | 1992-02-06 | 2002-12-11 | Schering Corporation | Monoclonal antibody, hybridoma, polypeptide and process for preparing thereof, isolated DNA, recombinant vector, host cell, humanized antibody and pharmaceutical composition |
US5593874A (en) * | 1992-03-19 | 1997-01-14 | Monsanto Company | Enhanced expression in plants |
ATE260971T1 (en) * | 1992-04-01 | 2004-03-15 | Univ Rockefeller | METHOD FOR THE IN VITRO CULTIVATION OF DENDRITIC PRECURSOR CELLS AND THEIR USE FOR IMMUNOGENIC PRODUCTION |
DK0615451T3 (en) * | 1992-05-26 | 2006-04-24 | Immunex Corp | Hitherto unknown cytokine that binds to CD30 |
CA2134773A1 (en) * | 1992-06-04 | 1993-12-09 | Robert J. Debs | Methods and compositions for in vivo gene therapy |
US5614184A (en) * | 1992-07-28 | 1997-03-25 | New England Deaconess Hospital | Recombinant human erythropoietin mutants and therapeutic methods employing them |
CA2142007C (en) * | 1992-08-11 | 2007-10-30 | Robert Glen Urban | Immunomodulatory peptides |
DE4228839A1 (en) | 1992-08-29 | 1994-03-03 | Behringwerke Ag | Methods for the detection and determination of mediators |
DE69232604T2 (en) * | 1992-11-04 | 2002-11-07 | City Of Hope Duarte | ANTIBODY CONSTRUCTS |
EP1757694A3 (en) * | 1992-11-05 | 2008-02-27 | Sloan Kettering Institute For Cancer Research | Prostate-specific membrane antigen |
US5738849A (en) * | 1992-11-24 | 1998-04-14 | G. D. Searle & Co. | Interleukin-3 (IL-3) variant fusion proteins, their recombinant production, and therapeutic compositions comprising them |
US5543297A (en) * | 1992-12-22 | 1996-08-06 | Merck Frosst Canada, Inc. | Human cyclooxygenase-2 cDNA and assays for evaluating cyclooxygenase-2 activity |
US6096331A (en) * | 1993-02-22 | 2000-08-01 | Vivorx Pharmaceuticals, Inc. | Methods and compositions useful for administration of chemotherapeutic agents |
CA2161095A1 (en) * | 1993-04-20 | 1994-10-27 | Solis Therapeutics, Inc. | Methods and materials for treatment of individuals infected with intracellular infectious agents |
US5759551A (en) * | 1993-04-27 | 1998-06-02 | United Biomedical, Inc. | Immunogenic LHRH peptide constructs and synthetic universal immune stimulators for vaccines |
JPH08509614A (en) * | 1993-04-29 | 1996-10-15 | アボツト・ラボラトリーズ | Erythropoietin analog compositions and methods |
US5554512A (en) | 1993-05-24 | 1996-09-10 | Immunex Corporation | Ligands for flt3 receptors |
CA2125763C (en) * | 1993-07-02 | 2007-08-28 | Maurice Kent Gately | P40 homodimer of interleukin-12 |
GB2280171B (en) * | 1993-07-22 | 1996-12-18 | Cargo Unit Containers Ltd | Improvments in or relating to freight containers |
CN1057534C (en) | 1993-08-17 | 2000-10-18 | 柯瑞英-艾格公司 | Erythropoietin analogs |
JP4115517B2 (en) * | 1994-01-25 | 2008-07-09 | エラン ファーマシューティカルズ, インコーポレイテッド | Humanized antibody against leukocyte adhesion molecule VLA-4 |
US5837682A (en) * | 1996-03-08 | 1998-11-17 | The Children's Medical Center Corporation | Angiostatin fragments and method of use |
US5639725A (en) * | 1994-04-26 | 1997-06-17 | Children's Hospital Medical Center Corp. | Angiostatin protein |
AU692865B2 (en) * | 1994-04-26 | 1998-06-18 | Children's Medical Center Corporation | Angiostatin and method of use for inhibition of angiogenesis |
CU22615A1 (en) | 1994-06-30 | 2000-02-10 | Centro Inmunologia Molecular | PROCEDURE FOR OBTAINING LESS IMMUNOGENIC MONOCLONAL ANTIBODIES. MONOCLONAL ANTIBODIES OBTAINED |
US6429199B1 (en) * | 1994-07-15 | 2002-08-06 | University Of Iowa Research Foundation | Immunostimulatory nucleic acid molecules for activating dendritic cells |
US6309853B1 (en) | 1994-08-17 | 2001-10-30 | The Rockfeller University | Modulators of body weight, corresponding nucleic acids and proteins, and diagnostic and therapeutic uses thereof |
US5541087A (en) * | 1994-09-14 | 1996-07-30 | Fuji Immunopharmaceuticals Corporation | Expression and export technology of proteins as immunofusins |
US6309636B1 (en) * | 1995-09-14 | 2001-10-30 | Cancer Research Institute Of Contra Costa | Recombinant peptides derived from the Mc3 anti-BA46 antibody, methods of use thereof, and methods of humanizing antibody peptides |
PT706799E (en) | 1994-09-16 | 2002-05-31 | Merck Patent Gmbh | IMUNOCONJUGADOS II |
US5935821A (en) * | 1995-01-17 | 1999-08-10 | Board Of Trustees Of The University Of Kentucky | Polynucleotides related to monoclonal antibody 1A7 and use for the treatment of melanoma and small cell carcinoma |
US6086875A (en) * | 1995-01-17 | 2000-07-11 | The Brigham And Women's Hospital, Inc. | Receptor specific transepithelial transport of immunogens |
US6485726B1 (en) | 1995-01-17 | 2002-11-26 | The Brigham And Women's Hospital, Inc. | Receptor specific transepithelial transport of therapeutics |
US5552524A (en) * | 1995-01-31 | 1996-09-03 | Eli Lilly And Company | Anti-obesity proteins |
US5691309A (en) * | 1995-01-31 | 1997-11-25 | Eli Lilly And Company | Anti-obesity proteins |
US5891680A (en) * | 1995-02-08 | 1999-04-06 | Whitehead Institute For Biomedical Research | Bioactive fusion proteins comprising the p35 and p40 subunits of IL-12 |
ES2225874T3 (en) * | 1995-03-10 | 2005-03-16 | Genentech, Inc. | RECEIVER ACTIVATION THROUGH GAS. |
US5719266A (en) * | 1995-03-17 | 1998-02-17 | Eli Lilly And Company | Anti-obesity proteins |
US6281010B1 (en) * | 1995-06-05 | 2001-08-28 | The Trustees Of The University Of Pennsylvania | Adenovirus gene therapy vehicle and cell line |
CA2225852C (en) * | 1995-06-30 | 2009-04-14 | Mochida Pharmaceutical Co., Ltd. | Anti-fas ligand antibody and assay method using the anti-fas ligand antibody |
WO1997002004A2 (en) * | 1995-06-30 | 1997-01-23 | Eli Lilly And Company | Methods for treating diabetes |
US6406689B1 (en) * | 1995-10-03 | 2002-06-18 | Frank W. Falkenberg | Compositions and methods for treatment of tumors and metastatic diseases |
US5854205A (en) | 1995-10-23 | 1998-12-29 | The Children's Medical Center Corporation | Therapeutic antiangiogenic compositions and methods |
US5723125A (en) * | 1995-12-28 | 1998-03-03 | Tanox Biosystems, Inc. | Hybrid with interferon-alpha and an immunoglobulin Fc linked through a non-immunogenic peptide |
US6080409A (en) * | 1995-12-28 | 2000-06-27 | Dendreon Corporation | Immunostimulatory method |
US6750334B1 (en) * | 1996-02-02 | 2004-06-15 | Repligen Corporation | CTLA4-immunoglobulin fusion proteins having modified effector functions and uses therefor |
US5834597A (en) * | 1996-05-20 | 1998-11-10 | Protein Design Labs, Inc. | Mutated nonactivating IgG2 domains and anti CD3 antibodies incorporating the same |
CN1136197C (en) * | 1996-05-30 | 2004-01-28 | 霍夫曼-拉罗奇有限公司 | Novel pyridajinone derivatives |
US5922685A (en) * | 1996-06-05 | 1999-07-13 | Powderject Vaccines, Inc. | IL-12 gene therapy of tumors |
AU725329B2 (en) * | 1996-09-02 | 2000-10-12 | Ko Okumura | Humanized immunoglobulin reacting specifically with Fas ligand or active fragments thereof and region inducing apoptosis originating in Fas ligand |
ATE218143T1 (en) * | 1996-09-03 | 2002-06-15 | Gsf Forschungszentrum Umwelt | USE OF BI- AND TRISPECIFIC ANTIBODIES TO INDUCE TUMOR IMMUNITY |
US6417337B1 (en) | 1996-10-31 | 2002-07-09 | The Dow Chemical Company | High affinity humanized anti-CEA monoclonal antibodies |
US5994104A (en) * | 1996-11-08 | 1999-11-30 | Royal Free Hospital School Of Medicine | Interleukin-12 fusion protein |
US6100387A (en) * | 1997-02-28 | 2000-08-08 | Genetics Institute, Inc. | Chimeric polypeptides containing chemokine domains |
US6277375B1 (en) | 1997-03-03 | 2001-08-21 | Board Of Regents, The University Of Texas System | Immunoglobulin-like domains with increased half-lives |
EP0973550B1 (en) * | 1997-04-11 | 2002-10-09 | G.D. SEARLE & CO. | Antagonistic anti-avb3 integrin antibodies |
CA2290485C (en) * | 1997-05-21 | 2008-08-05 | Biovation Limited | Method for the production of non-immunogenic proteins |
ATE267215T1 (en) | 1997-12-08 | 2004-06-15 | Lexigen Pharm Corp | HETERODIMARY FUSION PROTEINS FOR USE FOR TARGETED IMMUNTHERAPY AND GENERAL IMMUNE EXCITATION |
US20030105294A1 (en) | 1998-02-25 | 2003-06-05 | Stephen Gillies | Enhancing the circulating half life of antibody-based fusion proteins |
EP1071469A2 (en) * | 1998-04-17 | 2001-01-31 | Lexigen Pharmaceuticals Corp. | Enhancement of antibody-cytokine fusion protein mediated immune responses by co-administration with prostaglandin inhibitor |
WO1999054484A1 (en) | 1998-04-20 | 1999-10-28 | The Regents Of The University Of California | Modified immunoglobulin molecules and methods for use thereof |
WO1999058662A1 (en) | 1998-05-14 | 1999-11-18 | Merck Patent Gmbh | Fused protein |
US6620382B1 (en) * | 1998-05-22 | 2003-09-16 | Biopheresis Technologies, Llc. | Method and compositions for treatment of cancers |
US20020142374A1 (en) | 1998-08-17 | 2002-10-03 | Michael Gallo | Generation of modified molecules with increased serum half-lives |
CA2339331C (en) * | 1998-08-25 | 2011-03-01 | Lexigen Pharmaceuticals Corporation | Expression and export of angiogenesis inhibitors as immunofusins |
US6646113B1 (en) | 1998-09-17 | 2003-11-11 | The Trustees Of The University Of Pennsylvania | Nucleic acid molecule encoding human survival of motor neuron-interacting protein 1 (SIP1) deletion mutants |
US6335176B1 (en) * | 1998-10-16 | 2002-01-01 | Pharmacopeia, Inc. | Incorporation of phosphorylation sites |
ES2278463T3 (en) | 1998-12-08 | 2007-08-01 | Biovation Limited | METHOD TO REDUCE THE IMMUNOGENICITY OF PROTEINS. |
KR20020007287A (en) * | 1999-01-07 | 2002-01-26 | 추후보정 | EXPRESSION AND EXPORT OF ANTI-OBESITY PROTEINS AS Fc FUSION PROTEINS |
EP1185559A2 (en) | 1999-04-28 | 2002-03-13 | Board Of Regents, The University Of Texas System | Compositions and methods for cancer treatment by selectively inhibiting vegf |
KR100922809B1 (en) | 1999-05-06 | 2009-10-21 | 웨이크 포리스트 유니버시티 | Compositions And Methods For Identifying Antigens Which Elicit An Immune Response |
US6348192B1 (en) * | 1999-05-11 | 2002-02-19 | Bayer Corporation | Interleukin-2 mutein expressed from mammalian cells |
EP1187852B1 (en) * | 1999-05-19 | 2007-08-08 | EMD Lexigen Research Center Corp. | EXPRESSION AND EXPORT OF INTERFERON-ALPHA PROTEINS AS Fc FUSION PROTEINS |
JO2291B1 (en) * | 1999-07-02 | 2005-09-12 | اف . هوفمان لاروش ايه جي | Erythopintin derivatives |
CZ299516B6 (en) * | 1999-07-02 | 2008-08-20 | F. Hoffmann-La Roche Ag | Erythropoietin glycoprotein conjugate, process for its preparation and use and pharmaceutical composition containing thereof |
WO2001010912A1 (en) | 1999-08-09 | 2001-02-15 | Lexigen Pharmaceuticals Corp. | Multiple cytokine-antibody complexes |
WO2001023573A1 (en) * | 1999-09-30 | 2001-04-05 | Kyowa Hakko Kogyo Co., Ltd. | Human type complementation-determining domain transplanted antibody against ganglioside gd2 and derivative of this antibody |
CA2399832C (en) | 2000-02-11 | 2011-09-20 | Stephen D. Gillies | Enhancing the circulating half-life of antibody-based fusion proteins |
PT1257297E (en) * | 2000-02-24 | 2006-12-29 | Philogen Spa | Compositions and methods for treatemnt of angiogenesis in pathological lesions |
US6586398B1 (en) * | 2000-04-07 | 2003-07-01 | Amgen, Inc. | Chemically modified novel erythropoietin stimulating protein compositions and methods |
MXPA02011016A (en) * | 2000-05-12 | 2004-03-16 | Neose Technologies Inc | In vitro. |
ES2288967T3 (en) * | 2000-06-29 | 2008-02-01 | Merck Patent Gmbh | REINFORCEMENT OF IMMUNE ANSWERS MEDIATED BY THE ANTIBODY-CYTOKIN FUSION PROTEIN THROUGH THE TREATMENT COMBINED BY AGENTS THAT IMPROVE THE INCORPORATION OF IMMUNOCITOQUINE. |
JP2004525621A (en) * | 2001-01-18 | 2004-08-26 | メルク パテント ゲゼルシャフト ミット ベシュレンクテル ハフトング | Bifunctional fusion protein having glucocerebrosidase activity |
JP2004532620A (en) * | 2001-02-19 | 2004-10-28 | メルク パテント ゲゼルシャフト ミット ベシュレンクテル ハフトング | Artificial proteins with reduced immunogenicity |
CA2440221C (en) | 2001-03-07 | 2013-02-05 | Merck Patent Gesellschaft Mit Beschraenkter Haftung | Expression technology for proteins containing a hybrid isotype antibody moiety |
WO2002079415A2 (en) | 2001-03-30 | 2002-10-10 | Lexigen Pharmaceuticals Corp. | Reducing the immunogenicity of fusion proteins |
BR0209177A (en) * | 2001-05-03 | 2004-10-05 | Merck Patent Gmbh | Recombinant tumor specific antibody and use |
ES2381025T3 (en) | 2001-12-04 | 2012-05-22 | Merck Patent Gmbh | Immunocytokines with modulated selectivity |
US6996009B2 (en) * | 2002-06-21 | 2006-02-07 | Micron Technology, Inc. | NOR flash memory cell with high storage density |
ES2346205T3 (en) | 2002-12-17 | 2010-10-13 | Merck Patent Gmbh | HUMANIZED ANTIBODY (H14.18) OF ANTIBODY 14.18 OF MOUSE THAT LINKS TO GD2 AND ITS FUSION WITH IL-2. |
US9395802B2 (en) | 2014-05-22 | 2016-07-19 | Via Alliance Semiconductor Co., Ltd. | Multi-core data array power gating restoral mechanism |
-
2003
- 2003-12-16 ES ES03795898T patent/ES2346205T3/en not_active Expired - Lifetime
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US7767405B2 (en) | 2010-08-03 |
JP4494977B2 (en) | 2010-06-30 |
RU2005119305A (en) | 2006-01-20 |
CA2510180A1 (en) | 2004-07-01 |
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ATE471946T1 (en) | 2010-07-15 |
CN1726227A (en) | 2006-01-25 |
DK1572748T3 (en) | 2010-08-23 |
DE60333121D1 (en) | 2010-08-05 |
PT1572748E (en) | 2010-09-28 |
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AU2003298187A1 (en) | 2004-07-09 |
WO2004055056A1 (en) | 2004-07-01 |
BR0317376A (en) | 2005-11-16 |
RU2366664C2 (en) | 2009-09-10 |
KR101086660B1 (en) | 2011-11-24 |
US20040203100A1 (en) | 2004-10-14 |
US7169904B2 (en) | 2007-01-30 |
JP2006521085A (en) | 2006-09-21 |
AU2003298187B2 (en) | 2010-09-16 |
EP1572748A1 (en) | 2005-09-14 |
BRPI0317376B1 (en) | 2019-12-03 |
US20100210831A1 (en) | 2010-08-19 |
US20070059282A1 (en) | 2007-03-15 |
CN100432105C (en) | 2008-11-12 |
KR20050085775A (en) | 2005-08-29 |
BRPI0317376B8 (en) | 2021-05-25 |
EP1572748B1 (en) | 2010-06-23 |
MXPA05006384A (en) | 2005-08-29 |
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