Anti-CD84 antibodies and uses thereof

Abstract

Provided herein, inter alia, are antibodies (e.g., humanized antibodies, chimeric antibodies, monoclonal antibodies, antibody fragments (e.g., scFv)) that bind to cluster of differentiation 84 (CD84) with high efficiency and specificity. The antibodies provided herein include novel light chain domain CDRs and heavy chain domain CDRs and framework regions, and are useful, inter alia, in the treatment of cancer and other CD84-related diseases.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This international application claims the benefit of priority under 35 U.S.C. §119(e) to U.S. patent application No. 63 / 342,521, filed on May 16, 2022, which is hereby incorporated by reference in its entirety and for all purposes.

[0003] References to sequence listings

[0004] The entire text of the sequence listing written in the file 048440-750001US_SL_ST26.xml, created on May 14, 2023, 102,132 bytes, machine format for IBM-PC, using MS Windows operating system, is hereby incorporated by reference. Background Art

[0005] There is a need in the art for anti-CD84 antibodies to act on the tumor microenvironment, generate a favorable immune response and subsequent tumor eradication. Provided herein, inter alia, are compositions and methods that address these and other needs in the art. Summary of the invention

[0006] In one aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:75, CDR H2 as shown in SEQ ID NO:76, and CDR H3 as shown in SEQ ID NO:77; and wherein the light chain variable domain comprises CDRL1 as shown in SEQ ID NO:78, CDR L2 as shown in SEQ ID NO:79, and CDR L3 as shown in SEQ ID NO:80.

[0007] In another aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:19, CDRH2 as shown in SEQ ID NO:20, and CDR H3 as shown in SEQ ID NO:21; and wherein the light chain variable domain comprises CDR L1 as shown in SEQ ID NO:22, CDR L2 as shown in SEQ ID NO:23, and CDR L3 as shown in SEQ ID NO:24.

[0008] In another aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:39, CDRH2 as shown in SEQ ID NO:40, and CDR H3 as shown in SEQ ID NO:41; and wherein the light chain variable domain comprises CDR L1 as shown in SEQ ID NO:42, CDR L2 as shown in SEQ ID NO:43, and CDR L3 as shown in SEQ ID NO:44.

[0009] In another aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:57, CDRH2 as shown in SEQ ID NO:58, and CDR H3 as shown in SEQ ID NO:59; and wherein the light chain variable domain comprises CDR L1 as shown in SEQ ID NO:60, CDR L2 as shown in SEQ ID NO:61, and CDR L3 as shown in SEQ ID NO:62.

[0010] In another aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:93, CDRH2 as shown in SEQ ID NO:94, and CDR H3 as shown in SEQ ID NO:95; and wherein the light chain variable domain comprises CDR L1 as shown in SEQ ID NO:96, CDR L2 as shown in SEQ ID NO:97, and CDR L3 as shown in SEQ ID NO:98.

[0011] In another aspect, an anti-CD84 antibody is provided, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:19, CDR H2 as shown in SEQ ID NO:20, and CDR H3 as shown in SEQ ID NO:21, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:22, CDR L2 as shown in SEQ ID NO:23, and CDR L3 as shown in SEQ ID NO:24.

[0012] In another aspect, an anti-CD84 antibody is provided, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:39, CDR H2 as shown in SEQ ID NO:40, and CDR H3 as shown in SEQ ID NO:41, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:42, CDR L2 as shown in SEQ ID NO:43, and CDR L3 as shown in SEQ ID NO:44.

[0013] In another aspect, an anti-CD84 antibody is provided, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:57, CDR H2 as shown in SEQ ID NO:58, and CDR H3 as shown in SEQ ID NO:59, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:60, CDR L2 as shown in SEQ ID NO:61, and CDR L3 as shown in SEQ ID NO:62.

[0014] In another aspect, an anti-CD84 antibody is provided, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:75, CDR H2 as shown in SEQ ID NO:76, and CDR H3 as shown in SEQ ID NO:77, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:78, CDR L2 as shown in SEQ ID NO:79, and CDR L3 as shown in SEQ ID NO:80.

[0015] In another aspect, an anti-CD84 antibody is provided, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:93, CDR H2 as shown in SEQ ID NO:94, and CDR H3 as shown in SEQ ID NO:95, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:96, CDR L2 as shown in SEQ ID NO:97, and CDR L3 as shown in SEQ ID NO:98.

[0016] In another aspect, an isolated nucleic acid encoding an anti-CD84 antibody as provided herein (including embodiments thereof) is provided.

[0017] In another aspect, a cell is provided, the cell comprising an anti-CD84 antibody as provided herein (including embodiments thereof), or a nucleic acid as provided herein (including embodiments thereof).

[0018] In another aspect, a pharmaceutical composition is provided, comprising a therapeutically effective amount of an antibody as provided herein (including embodiments thereof) and a pharmaceutically acceptable excipient.

[0019] In another aspect, a method of generating an antibody capable of binding to CD84 is provided, the method comprising immunizing a mammal with a peptide comprising the sequence of SEQ ID NO: 111.

[0020] In another aspect, a method of treating cancer in a subject in need thereof is provided, the method comprising administering to the subject a therapeutically effective amount of an anti-CD84 antibody as provided herein (including embodiments thereof) or a pharmaceutical composition as provided herein (including embodiments thereof), thereby treating the cancer in the subject. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 Recombinant CD84 protein constructs are exemplified in which the extracellular domain (ECD) of human CD84 is fused to mouse IgG2a.Fc (left panel) or human IgA1.Fc (right panel).

[0022] Figure 2 Illustrated is the purification and characterization of recombinant CD84 protein fused to IgG2a (upper panel) or IgA1 (lower panel).

[0023] Figure 3 Exemplary flow cytometry data are illustrated showing that 16 CD84 clones were able to bind human CD84 on the cell surface of a mouse myeloma cell line (ECACC).

[0024] Figure 4 Exemplary data from a flow cytometry experiment testing six CD84 clones (2H6, 4B11, 5C6, 7D2, 8A5, and 13D11) for their ability to bind to the human myeloma cell line HL-60, the T cell line Jurkat, the lymphoma cell line K-562, and the breast cell line SKBR3 (ATCC) are shown.

[0025] Figure 5 Exemplary data from ELISA experiments are illustrated showing that the 2H6 clone, 4B11 clone, 5C6 clone, 7D2 clone, 8A5 clone, and 13D11 clone are of Igγ1 and Igκ subtypes.

[0026] FIG. 6A to FIG. 6E Clone 5C6 ( Fig. 6A), clone 7D2( Figure 6B ), clone 8A5( Figure 6C ), clone 13D11( Fig. 6E )'s chimeric Fab purification and characterization. Fig. 6A is a chromatogram depicting the purification of chimeric Fab of clone 5C6 using size exclusion chromatography. Figure 6B is a chromatogram depicting the purification of chimeric Fab of clone 7D2 using size exclusion chromatography. Figure 6C is a chromatogram depicting purification of chimeric Fab of clone 8A5 using size exclusion chromatography. Fig.6D Shown are SDS-PAGE gels characterizing purified chimeric Fabs of clone 5C6, clone 8A5 and clone 7D2 under reducing (R) and non-reducing (NR) conditions. Fig. 6E are chromatograms depicting purification of chimeric Fab of clone 7D2 using size exclusion chromatography (left panel) and SDS-PAGE gels characterizing purified chimeric Fab of clone 13D11 under reducing (R) and non-reducing (NR) conditions.

[0027] 7A to 7D Shown are sensorgrams from an SPR experiment depicting the chimeric anti-CD84 Fab against clone 13D11 ( Fig. 7A ), clone 5C6( Figure 7B ), clone 7D2( Figure 7C ) and clone 8A5 ( Fig.7D ). Anti-HIS antibody was immobilized on the SPR biosensor chip, and anti-CD84 chimeric Fab was injected at concentrations of 300 nM, 100 nM, 30 nM, and 10 nM at 25°C.

[0028] Figure 8 Thermal stability data and melting point temperatures (Tm) of chimeric anti-CD84 Fabs of clone 5C6, clone 7D2, clone 8A5 and clone 13D11 are exemplified.

[0029] Fig. 9 Ribbon diagram depicting the atomic structure of murine chimeric anti-CD84 8A5 clone, 7D2 clone, and 5C6 clone derived from X-ray diffraction data.

[0030] FIG. 10A to FIG. 10F Shown are the purification and characterization of humanized anti-CD84 Fab and antibodies of the 4B11 clone and the 13D11 clone. Fig. 10A is a chromatogram depicting the purification of humanized anti-CD84 antibody 4B11 clone using size exclusion chromatography. Fig. 10B is a chromatogram depicting the purification of humanized anti-CD84 antibody 13D11 clone using size exclusion chromatography. Fig. 10CShown is an SDS-PAGE gel characterizing purified humanized anti-CD84 antibodies 4B11 clone and 13D11 clone under reducing (R) and non-reducing (NR) conditions. Fig. 10D is a chromatogram depicting the purification of humanized anti-CD84 4B11 clone Fab using size exclusion chromatography. Fig. 10E is a chromatogram depicting the purification of humanized anti-CD84 13D11 clone Fab using size exclusion chromatography. Fig.10F Shown are SDS-PAGE gels characterizing purified humanized anti-CD84 Fab of clone 4B11 and clone 13D11 under reducing (R) and non-reducing (NR) conditions.

[0031] FIG. 11A to FIG. 11B Illustrated is the expression of CD84 in samples from AML patients compared to healthy donors. Fig.11A is a scatter plot showing CD84 messenger RNA (mRNA) expression in the GSE9476 and GSE13159 datasets, which include samples from AML patients and healthy donors, highlighting increased CD84 expression in AML samples, especially in myelomonocytic and monocytic leukemia patients with t(11q23) and inv(16) abnormalities; GSE9476 (healthy donors n=38; AML n=26); GSE13159 (healthy donors n=73; AML n=541). Fig. 11B Is a violin plot showing CD84 surface protein expression in healthy donors (n=4), AML patient samples (n=4) and AML cell lines (n=6) analyzed by flow cytometry, highlighting that CD84 is highly expressed in AML primary patient cells and AML cell lines relative to healthy donors. The Y axis represents the mean fluorescence intensity (MFI) of CD84 relative to isotype. Fig. 11C is a survival curve showing that high CD84 expression is associated with shorter overall survival (OS) in AML patients (GSE10358). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001

[0032] FIG. 12A to FIG. 12C Shown are the results from knocking down AML cell lines using short hairpin RNA (sh) Fig. 12A ), primary patient AML cells ( Fig. 12B ) and THP1-xenograft mouse models ( Fig. 12C ) are exemplary data from experiments with CD84. Fig. 12A(Left panel) is a line graph representing cell proliferation analysis of THP1 cells and HEL cells transduced with shControl (shCtrl) or shCD84 lentiviral vectors, showing that CD84 knockout inhibited cell proliferation in AML cell lines. Error bars represent SEM from three independent experiments. Fig. 12A (Right panel) is a bar graph showing apoptosis analysis, indicating Annexin-APC / DAPI in THP1 cells and HEL cells transduced with shCtrl or shCD84 lentiviral vectors, indicating CD84 knockout-induced apoptosis in AML cells. Fig. 12B is a bar graph showing apoptosis analysis indicated by Annexin-APC / DAPI in two AML patient samples transduced with shCtrl or shCD84 lentiviral vectors, demonstrating that CD84 knockout induces apoptosis in AML primary patient cells. Error bars represent SEM from three technical experiments. Fig. 12C Bioluminescent imaging is shown, which shows the tumor burden in xenografted NSG mice at day 35 after transplantation of shCtrl or shCD84 transduced THP1-luciferase cells. Kaplan-Meier analysis of the survival of NSG mice (n=5 / group) transplanted with THP1-luciferase cells (shCtrl or shCD84). Relative to control mice, attenuated tumor burden and prolonged survival were observed in mice transplanted with CD84 knockout cells. *p<0.05, **p<0.01, ***p<0.001.

[0033] FIG. 13A to FIG. 13E Exemplary data from experiments showing that CD84 knockout inhibits MLL-AF9 cell growth (in vitro) and attenuates cell engraftment (in vivo) are illustrated. Fig.13A The workflow for generating MLL-AF9 AML cells with CD84 knockout is shown. Briefly, c-kit positive bone marrow cells collected from C57BL / 6 mice (CD45.2) were retrovirally transduced with MLL-AF9 and then lentivirally transduced with shCtrl, shCD84-1 or shCD84-2. Cells were used for cell proliferation analysis, colony formation assays. MLL-AF9 transduced cells were also transplanted into irradiated recipient cells for leukemogenesis. Fig. 13B is a line graph representing cell proliferation analysis of MLL-AF9 cells transduced with shCtrl or shCD84 lentiviral vectors, indicating that CD84 knockout inhibited cell proliferation in MLL-AF9 AML cells. Fig. 13CFigures and bar graphs show MLL-AF9 AML cells transduced with shCtrl, shCD84-1 or shCD84-2 and then plated for colony formation assays. These results indicate that CD84 knockout inhibits AML cell colony formation ability. Fig.13D Exemplary data from mouse experiments are illustrated, showing that CD84 knockout inhibits AML engraftment in the MLL-AF9 xenograft mouse model. The dot plot shows engraftment of donor AML mouse cells in the bone marrow (mouse CD45.2). Fig.13E MLL-AF9 xenografts with CD84 knockout cells were shown to reduce splenomegaly. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

[0034] FIG. 14A to FIG. 14B Exemplary data from an experiment showing that CD84 knockout reduces AML engraftment in an AML patient-derived xenograft (PDX) mouse model are illustrated. The dot plots show lower AML engraftment (human CD45 / CD33 positivity ratio) in the bone marrow, spleen, and peripheral blood (PB) of NSG mice transplanted with AML primary cells transfected with shCD84 lentivirus relative to the shCtrl group ( Fig.14A ) and decreased spleen weight ( Fig. 14B ). *p<0.05, **p<0.01.

[0035] FIG. 15A to FIG. 15B Exemplary data from experiments in which AML cells cultured using total PBMCs or NK cells from healthy donors were treated with anti-CD84 antibodies are shown. Fig.15A Shown are shifted histograms showing CD84 expression detected by flow cytometry in CD84 high-expressing cells THP1 and CD84 low / null-expressing cells MM1S (upper panel) and bar graphs showing that anti-CD84 monoclonal antibody clones (5C6, 7D2 and 8A5) induce antibody-dependent cellular cytotoxicity (ADCC) in THP1 (middle panel) and MM1S cells (lower panel), highlighting that the anti-CD84 7D2 antibody exhibits high anti-leukemia activity and specificity. Fig. 15B Line graphs illustrating that anti-CD84 7D2 antibody induces ADCC of THP1-luciferase cells (target cells) in the presence of total PBMC cells (upper and middle panels) or NK cells (lower panel) as effector cells, as indicated by 7-ADD positivity. The results highlight that anti-CD84 antibody-induced ADCC is E:T ratio-dependent and antibody dose-dependent. Healthy donor PBMC (n=2); healthy donor NK cells (n=2).

[0036] FIG. 16A to FIG. 16BExemplary data from experiments showing that anti-CD84 antibody treatment (clone 7D2) reduced tumor burden and prolonged survival in THP1 and U937 xenograft models are illustrated. Fig.16A Bioluminescent imaging of tumor burden in THP1-xenografted NSG mice showing luciferase expression after anti-human IgG / anti-CD84 antibody treatment (clone 7D2) is shown. Kaplan-Meier analysis of survival of treated NSG mice (n=6 / group). The reduced tumor burden and prolonged survival time highlight that the anti-CD84 antibody (clone 7D2) showed a significant anti-leukemic effect. Fig. 16B Bioluminescent imaging of tumor burden in U937-xenografted NSG mice indicating luciferase expression after anti-human IgG / anti-CD84 antibody treatment (clone 7D2) is shown. Kaplan-Meier analysis of survival of treated NSG mice (n=6, IgG group; n=7, anti-CD84 7D2 antibody group). The reduced tumor burden and prolonged survival time highlight that anti-CD84 antibody (clone 7D2) showed a significant anti-leukemic effect. **p<0.01, ***p<0.001.

[0037] Fig.17 Exemplary data from an experiment showing that anti-CD84 antibody treatment (clone 7D2) reduces AML implantation in an AML patient-derived xenograft (PDX) mouse model are illustrated. The dot plots show AML implantation (human CD45 / CD33 positive rate) in the bone marrow, spleen, and peripheral blood of NSG mice (n=6 / group) transplanted with AML PDX treated with anti-human IgG or anti-CD84 antibody (clone 7D2), as well as spleen weight. The results indicate that anti-CD84 antibody (clone 7D2) significantly inhibits AML progression in vivo. Error bars represent SEM. **p<0.01, ***p<0.001.

[0038] Fig.18 Statistics from data collection and refinement of the 8A5-CD84 complex are shown.

[0039] Fig.19 The crystal structure of the 8A5-CD84 complex (using X-ray crystallography) is shown. Note that the dimer interface is generated by the symmetry elements of the crystal.

[0040] FIG. 20A to FIG. 20F Data showing co-elution of anti-CD84 Fab binding using fast protein liquid chromatography (FPLC) data. FIG. 20A to FIG. 20C The chromatogram of CD84 mixed with a single Fab is shown. The shift to earlier volumes indicates the formation of a Fab / CD84 complex. FIG. 20D to FIG. 20FA chromatogram is shown using a mixture of two Fabs and CD84. Note that the X-axis is different - Fig.20D is from 10ml to 15ml, and FIG. 20E to FIG. 20F It is 8mL to 15mL. Fig.20D In , the displacement of the complex is similar to that of the single Fab trace (about 11 ml), indicating that 7D2 and 5C6 bind to a common or close epitope, such that one sterically blocks the other from binding to CD84. FIG. 20E to FIG. 20F In the ELISA, there is a substantial left shift (early elution time, approximately 9.5 ml), indicating binding to different, non-overlapping epitopes. DETAILED DESCRIPTION

[0041] definition

[0042] Although various embodiments and aspects of the present invention are shown and described herein, it will be apparent to those skilled in the art that such embodiments and aspects are provided by way of example only. Without departing from the present invention, those skilled in the art will be able to make numerous changes, modifications and substitutions. It should be understood that various alternatives to the embodiments of the present invention described herein may be employed in the implementation of the present invention.

[0043] The section headings used herein are for organizational purposes only and should not be construed in any way as limiting the subject matter described. All documents or portions of documents cited in this application, including but not limited to patents, patent applications, articles, books, manuals, and treatises, are hereby expressly incorporated herein by reference in their entirety for any purpose.

[0044] Abbreviations used herein have their conventional meanings within the chemical and biological arts.Chemical structures and formulae set forth herein are constructed according to standard rules of chemical valency known in the chemical arts.

[0045] Unless otherwise defined, the technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art. See, for example, Singleton et al., DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY 2nd Edition, J. Wiley & Sons (New York, NY 1994); Sambrook et al., MOLECULAR CLONING, A LABORATORY MANUAL, Cold Springs Harbor Press (Cold Springs Harbor, NY 1989). Any method, device and material similar to or equivalent to those described herein may be used to implement the present invention. The following definitions are provided to facilitate understanding of certain terms frequently used herein, and are not intended to limit the scope of the present disclosure.

[0046] Although various embodiments and aspects of the present invention are shown and described herein, it will be apparent to those skilled in the art that such embodiments and aspects are provided by way of example only. Without departing from the present invention, those skilled in the art will be able to make numerous changes, modifications and substitutions. It should be understood that various alternatives to the embodiments of the present invention described herein may be employed in the implementation of the present invention.

[0047] The section headings used herein are for organizational purposes only and should not be construed in any way as limiting the subject matter described. All documents or portions of documents cited in this application, including but not limited to patents, patent applications, articles, books, manuals, and treatises, are hereby expressly incorporated herein by reference in their entirety for any purpose.

[0048] Abbreviations used herein have their conventional meanings within the chemical and biological arts.Chemical structures and formulae set forth herein are constructed according to standard rules of chemical valency known in the chemical arts.

[0049] Unless otherwise defined, the technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art. See, for example, Singleton et al., DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY 2nd Edition, J. Wiley & Sons (New York, NY 1994); Sambrook et al., MOLECULAR CLONING, A LABORATORY MANUAL, Cold Springs Harbor Press (Cold Springs Harbor, NY 1989). Any method, device and material similar to or equivalent to those described herein may be used to implement the present invention. The following definitions are provided to facilitate understanding of certain terms frequently used herein, and are not intended to limit the scope of the present disclosure.

[0050] "Nucleic acid" refers to nucleotides (e.g., deoxyribonucleotides or ribonucleotides) and their polymers, or their complements, in single-stranded, double-stranded or multi-stranded forms; or nucleosides (e.g., deoxyribonucleosides or ribonucleosides). In an embodiment, "nucleic acid" does not include nucleosides. The terms "polynucleotide", "oligonucleotide", etc. refer to the linear sequence of nucleotides in the usual and customary sense. The term "nucleoside" refers to a glycosylamine containing a nucleobase and a pentose (ribose or deoxyribose) in the usual and customary sense. Non-limiting examples of nucleosides include cytidine, uridine, adenosine, guanosine, thymidine and inosine. The term "nucleotide" refers to a single unit of a polynucleotide in the usual and customary sense, that is, a monomer. Nucleotides can be ribonucleotides, deoxyribonucleotides or modified forms thereof. Examples of polynucleotides contemplated herein include single-stranded and double-stranded DNA, single-stranded and double-stranded RNA, and hybrid molecules with a mixture of single-stranded and double-stranded DNA and RNA. Examples of nucleic acids, such as polynucleotides encompassed herein include any type of RNA (e.g., mRNA, siRNA, miRNA, and guide RNA) and any type of DNA, genomic DNA, plasmid DNA, and minicircle DNA, and any fragments thereof. In the case of polynucleotides, the term "duplex" refers to a double strand in the usual and customary sense. Nucleic acids can be straight or branched. For example, nucleic acids can be linear nucleotide chains or nucleic acids can be branched, such as whereby the nucleic acid comprises arms or branches of one or more nucleotides. Optionally, branched nucleic acids repeatedly branch to form higher order structures, such as dendrimers, etc.

[0051] Nucleic acids, including, for example, nucleic acids having a phosphorothioate backbone, can include one or more reactive moieties. As used herein, the term reactive moiety includes any group that can react with another molecule (e.g., a nucleic acid or a polypeptide) through covalent, non-covalent, or other interactions. For example, a nucleic acid can include an amino acid reactive moiety that reacts with an amino acid on a protein or polypeptide through covalent, non-covalent, or other interactions.

[0052] The term also encompasses nucleic acids containing known nucleotide analogs or modified backbone residues or connections, which are synthetic, naturally occurring and non-naturally occurring, and which have binding properties similar to reference nucleic acids and which are metabolized in a manner similar to reference nucleotides. Examples of such analogs include, but are not limited to, phosphodiester derivatives, including, for example, phosphoramidates, phosphorodiamidates, phosphorothioates (also referred to as the thiophosphates of the oxygen in double bond sulfur replacement phosphates), phosphorodithioates, phosphonocarboxylic acids, phosphonocarboxylates, phosphonoacetic acid, phosphonoformic acid, methyl phosphonate, phosphonic acid boron or O-methyl phosphoramidate linking bonds (see Eckstein, OLIGONUCLEOTIDES AND ANALOGUES:APRACTICAL APPROACH, Oxford University Press), and modifications to nucleotide bases, such as in 5-methylcytidine or pseudouridine; and peptide nucleic acid backbones and linking bonds. Other analog nucleic acids include analog nucleic acids with the following items: positive backbone; non-ionic backbone, modified sugar and non-ribose backbone (e.g., phosphorodiamidate morpholino oligonucleotides or locked nucleic acids (LNA) known in the art), including the backbones described in U.S. Patents 5,235,033 and 5,034,506 and ASC Symposium Series 580, Chapters 6 and 7, CARBOHYDRATE MODIFICATIONS IN ANTISENSE RESEARCH, Sanghui & Cook editors. Nucleic acids containing one or more carbocyclic sugars are also included in a definition of nucleic acid. For a variety of reasons, modifications of the ribose-phosphate backbone can be performed, for example, to increase the stability and half-life of such molecules in physiological environments or as probes on biochips. Mixtures of naturally occurring nucleic acids and analogs can be prepared; alternatively, mixtures of different nucleic acid analogs and mixtures of naturally occurring nucleic acids and analogs can be prepared. In an embodiment, the internucleotide linkage in DNA is a phosphodiester, a phosphodiester derivative, or a combination of the two.

[0053] Nucleic acid can comprise non-specific sequence.As used herein, term " non-specific sequence " refers to the nucleic acid sequence containing a series of residues that are not designed to be complementary to any other nucleic acid sequence or are only partially complementary to any other nucleic acid sequence.For example, non-specific nucleic acid sequence is the nucleic acid residue sequence that does not play the inhibitory nucleic acid effect when contacting with cell or organism.

[0054] A polynucleotide is generally composed of a specific sequence of four nucleotide bases: adenine (A); cytosine (C); guanine (G); and thymine (T) (when the polynucleotide is RNA, uracil (U) represents thymine (T)). Therefore, the term "polynucleotide sequence" is an alphabetic representation of a polynucleotide molecule; alternatively, the term can be applied to the polynucleotide molecule itself. This alphabetic representation can be entered into a database in a computer with a central processing unit and used for bioinformatics applications such as functional genomics and homology searches. A polynucleotide may optionally include one or more non-standard nucleotides, nucleotide analogs and / or modified nucleotides.

[0055] As used herein, the term "complement" refers to a nucleotide (e.g., RNA or DNA) or a nucleotide sequence that can base pair with a complementary nucleotide or nucleotide sequence. As described herein and as known in the art, the complementary (matching) nucleotide of adenosine is thymidine, and the complementary (matching) nucleotide of guanine is cytosine. Therefore, the complement may include a nucleotide sequence that base pairs with the corresponding complementary nucleotide of the second nucleic acid sequence. The nucleotides of the complement may partially or completely match the nucleotides of the second nucleic acid sequence. When the nucleotides of the complement fully match each nucleotide of the second nucleic acid sequence, the complement forms a base pair with each nucleotide of the second nucleic acid sequence. When the nucleotides of the complement partially match the nucleotides of the second nucleic acid sequence, only some nucleotides of the complement form a base pair with the nucleotides of the second nucleic acid sequence. Examples of complementary sequences include coding and non-coding sequences, wherein the non-coding sequence contains the complementary nucleotides of the coding sequence, thereby forming the complement of the coding sequence. Another example of a complementary sequence is a sense sequence and an antisense sequence, wherein the sense sequence contains the complementary nucleotides of the antisense sequence, thereby forming the complement of the antisense sequence.

[0056] As described herein, sequence complementarity can be partial, in which only some of the nucleic acids match based on base pairing, or complete, in which all nucleic acids match based on base pairing. Thus, two sequences that are complementary to each other can have a specified percentage of identical nucleotides (i.e., about 60% identity over a specified region, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identity).

[0057] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to naturally occurring amino acids. Naturally occurring amino acids are amino acids encoded by the genetic code and amino acids that are subsequently modified, for example, hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. Amino acid analogs refer to compounds having the same basic chemical structure as naturally occurring amino acids, i.e., an α carbon bound to a hydrogen, carboxyl, amino, and R groups, for example, homoserine, norleucine, methionine sulfoxide, methionine methylsulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as naturally occurring amino acids. Amino acid mimetics refer to compounds that have a structure that is different from the general chemical structure of an amino acid but function in a manner similar to naturally occurring amino acids. The terms "non-naturally occurring amino acids" and "non-natural amino acids" refer to amino acid analogs, synthetic amino acids, and amino acid mimetics that are not found in nature.

[0058] Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Likewise, nucleotides may be referred to by their commonly accepted single-letter codes.

[0059] The terms "polypeptide", "peptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues, which in embodiments may be conjugated to a moiety that does not consist of amino acids. The terms apply to amino acid polymers in which one or more amino acid residues is an artificial chemical mimetic of the corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. A "fusion protein" refers to a chimeric protein encoding two or more separate protein sequences that are recombinantly expressed as a single part.

[0060] The "position" of an amino acid or nucleotide base is represented by a number that sequentially identifies each amino acid (or nucleotide base) in the reference sequence according to its position relative to the N-terminus (or 5' end). Since deletions, insertions, truncations, fusions, etc. must be considered when determining the optimal alignment, the numbering of the amino acid residues in the test sequence determined by counting only from the N-terminus is not necessarily the same as the numbering of the corresponding position in the reference sequence. For example, in the case where the variant has a deletion relative to the aligned reference sequence, there will be no amino acid in the variant corresponding to the position at the deletion site in the reference sequence. In the case of an insertion in the aligned reference sequence, the insertion will not correspond to the numbered amino acid position in the reference sequence. In the case of truncations or fusions, there may be amino acid segments in the reference sequence or aligned sequence that do not correspond to any amino acid in the corresponding sequence.

[0061] When used in the context of numbering a given amino acid or polynucleotide sequence, the term "reference ... number" or "corresponding to" refers to the numbering of the residues of a given reference sequence when the given amino acid or polynucleotide sequence is compared to the reference sequence. An amino acid residue in a protein "corresponds" to a given residue when it occupies the same basic structural position in the protein as the given residue. One skilled in the art will immediately recognize the identity and position of residues corresponding to specific positions in proteins (e.g., EGFR) in other proteins with different numbering systems. For example, by performing a simple sequence alignment with a protein (e.g., EGFR), the identity and position of residues corresponding to specific positions of the protein are identified in other protein sequences aligned with the protein. For example, when the selected residue has the same basic spatial or other structural relationship with the glutamic acid at position 138, the selected residue in the selected protein corresponds to the glutamic acid at position 138. In some embodiments, where the selected protein is aligned to the protein for maximum homology, the position in the selected protein aligned with glutamic acid 138 will correspond to glutamic acid 138. Instead of a primary sequence alignment, a three-dimensional structural alignment can also be used, for example, in which the structures of the selected proteins are aligned for maximum correspondence with glutamic acid at position 138 and the overall structures compared. In this case, the amino acid that occupies the same essential position as glutamic acid 138 in the structural model is the residue corresponding to the glutamic acid 138 residue.

[0062] "Conservatively modified variants" apply to amino acids and nucleic acid sequences. With respect to specific nucleic acid sequences, "conservatively modified variants" refer to those nucleic acids encoding identical or substantially identical amino acid sequences. Due to the degeneracy of the genetic code, many nucleic acid sequences will encode any given protein. For example, the codons GCA, GCC, GCG, and GCU all encode the amino acid alanine. Therefore, at each position where alanine is specified by a codon, the codon can be changed to any corresponding codon described without changing the encoded polypeptide. Such nucleic acid variations are "silent variations," which are a type of conservatively modified variations. Each nucleic acid sequence encoding a polypeptide herein also describes each possible silent variation of the nucleic acid. The skilled person will recognize that each codon in the nucleic acid (except AUG, which is usually the only codon for methionine, and TGG, which is usually the only codon for tryptophan) can be modified to obtain functionally identical molecules. Therefore, each silent variation of a nucleic acid encoding a polypeptide is implicit in each described sequence.

[0063] With respect to amino acid sequences, one skilled in the art will recognize that a single substitution, deletion or addition of a nucleic acid, peptide, polypeptide or protein sequence that alters, adds or deletes a single amino acid or a small portion of amino acids in the encoding sequence is a "conservatively modified variant," wherein the alteration results in the substitution of an amino acid with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. Such conservatively modified variants are in addition to, and do not exclude, polymorphic variants, interspecies homologs and alleles of the present disclosure.

[0064] Each of the following eight groups contains amino acids that are conservative substitutions for one another:

[0065] 1) Alanine (A), glycine (G);

[0066] 2) Aspartic acid (D), glutamic acid (E);

[0067] 3) Asparagine (N), glutamine (Q);

[0068] 4) Arginine (R), Lysine (K);

[0069] 5) Isoleucine (I), leucine (L), methionine (M), valine (V);

[0070] 6) Phenylalanine (F), tyrosine (Y), tryptophan (W);

[0071] 7) serine (S), threonine (T); and

[0072] 8) Cysteine ​​(C), Methionine (M)

[0073] (See, e.g., Creighton, Proteins (1984)).

[0074] In the context of two or more nucleic acid or polypeptide sequences, the term "identical" or "identity" percentage refers to two or more sequences or subsequences of identical or specified percentages of amino acid residues or identical nucleotides (i.e., about 60% identity over a specified region, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity when compared and aligned for maximum correspondence over a comparison window or specified region) (e.g., NCBI website http: / / www.ncbi.nlm.nih.gov / BLAST / , etc.). Such sequences are then referred to as "substantially identical". This definition also refers to or may be applicable to the complementary sequence of a test sequence. The definition also includes sequences with deletions and / or additions, as well as sequences with substitutions. As described below, preferred algorithms can account for gaps, etc. Preferably, identity exists over a region that is at least about 25 amino acids or nucleotides in length, or more preferably over a region that is 50-100 amino acids or nucleotides in length.

[0075] The "percentage of sequence identity" is determined by comparing two optimally aligned sequences in a comparison window, wherein for optimal alignment of the two sequences, the portion of the polynucleotide or polypeptide sequence in the comparison window may include additions or deletions (i.e., gaps) compared to the reference sequence (which does not include additions or deletions). The percentage is calculated by determining the number of positions where the same nucleic acid base or amino acid residue occurs in the two sequences to obtain the number of matched positions, dividing the number of matched positions by the total number of positions in the comparison window and multiplying the result by 100 to obtain the percentage of sequence identity.

[0076] As used herein, "comparison window" includes reference to a fragment selected from, for example, the full-length sequence or any number of consecutive positions of 20 to 600, about 50 to about 200, or about 100 to about 150 amino acids or nucleotide sequences, wherein after the two sequences are optimally aligned, the sequence can be compared to a reference sequence of the same number of consecutive positions. Methods of sequence alignment for comparison are well known in the art. Optimal alignment of sequences for comparison can be performed by the local homology algorithm of Smith and Waterman (1970) Adv. Appl. Math. 2:482c, the homology alignment algorithm of Needleman and Wunsch (1970) J. Mol. Biol. 48:443, the search for similarity method of Pearson and Lipman (1988) Proc. Nat'l. Acad. Sci. USA 85:2444, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI), or by manual alignment and visual inspection (see, e.g., Ausubel et al., Current Protocols in Molecular Biology (1995 Supplement)).

[0077] Examples of algorithms suitable for determining percentages of sequence identity and sequence similarity are BLAST and BLAST 2.0 algorithms, which are described in Altschul et al. (1977) Nuc. Acids Res. 25: 3389-3402, and Altschul et al. (1990) J. Mol. Biol. 215: 403-410, respectively. Software for performing BLAST analysis is publicly available through the National Center for Biotechnology Information (http: / / www.ncbi.nlm.nih.gov / ). This algorithm involves first identifying high-scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence that match or satisfy certain positive threshold scores T when aligned with words of the same length in the database sequence. T is referred to as the adjacent word score threshold (Altschul et al., supra). These initial adjacent word hits serve as seeds for initiating searches to find longer HSPs containing them. The word hits are extended in both directions along each sequence until the cumulative alignment score can be increased. For nucleotide sequences, the cumulative score is calculated using the parameters M (reward score for a pair of matching residues; always > 0) and N (penalty score for mismatching residues; always < 0). For amino acid sequences, the cumulative score is calculated using a scoring matrix. Extension of the word hits in each direction is stopped when: the cumulative alignment score drops by an amount X from its maximum achieved value; the cumulative score approaches zero or lower due to the accumulation of one or more negative scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses by default a word length (W) of 11, an expectation (E) or 10, M=5, N=-4, and a comparison of both chains. For amino acid sequences, the BLASTP program uses as defaults a wordlength of 3 and an expectation (E) of 10 and the BLOSUM62 scoring matrix (see Henikoff and Henikoff (1989) Proc. Natl. Acad. Sci. USA 89:10915) with an alignment (B) of 50, an expectation (E) of 10, M=5, N=-4, and a comparison of both strands.

[0078] The BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90:5873-5787). One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability that a match between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered similar to a reference sequence if the smallest sum probability in a comparison of a test nucleic acid to a reference nucleic acid is less than about 0.2, more preferably less than about 0.01, and most preferably less than about 0.001.

[0079] As described below, an indication that two nucleic acid sequences or polypeptides are substantially identical is that the polypeptide encoded by the first nucleic acid cross-reacts immunologically with antibodies produced for the polypeptide encoded by the second nucleic acid. Therefore, a polypeptide is generally substantially identical to a second polypeptide, for example, wherein the two peptides differ only by conservative substitutions. Another indication that two nucleic acid sequences are substantially identical is that the two molecules or their complementary sequences hybridize to each other under stringent conditions, as described below. Another indication that two nucleic acid sequences are substantially identical is that the same primers can be used for amplification of the sequence.

[0080] "CD84" referred to herein includes any of a recombinant or naturally occurring form of a cluster of differentiation 84 (CD84) protein or a variant or homolog thereof, including a CD84 complex that mediates signal transduction and maintains CD84 complex activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the activity compared to the CD84 complex). In some aspects, the variant or homolog has at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., a 50, 100, 150 or 200 contiguous amino acid portion) compared to the naturally occurring CD84 protein in the CD84 complex. In an embodiment, the CD84 protein is substantially identical to the protein identified by UniProt reference number Q9UIB8 or a variant or homolog substantially identical thereto.

[0081] Antibodies are large, complex molecules (molecular weight of ~150,000 or about 1320 amino acids) with a complex internal structure. A natural antibody molecule consists of two identical pairs of polypeptide chains, each pair having one light chain and one heavy chain. Each light and heavy chain, in turn, consists of two regions: a variable ("V") region that binds to the target antigen, and a constant ("C") region that interacts with other components of the immune system. The light and heavy chain variable regions (also referred to herein as light chain variable (VL) domains and heavy chain variable (VH) domains, respectively) bind together in 3-dimensional space to form a variable region that binds to an antigen (e.g., a receptor on the surface of a cell). Within each light or heavy chain variable region, there are three short segments (average length of 10 amino acids) called complementarity determining regions ("CDRs"). The six CDRs in the antibody variable domain (three from the light chain and three from the heavy chain) fold together in 3-dimensional space to form the actual antibody binding site that docks on the target antigen. The positions and lengths of CDRs have been precisely defined by Kabat, E. et al., Sequences of Proteins of Immunological Interest, US Department of Health and Human Services, 1983, 1987. The portion of the variable region not included in the CDRs is called the framework ("FR"), which forms the environment of the CDRs.

[0082] As provided herein, "antibody variant" refers to a polypeptide that is capable of binding to an antigen and includes one or more domains (e.g., light chain variable domain, heavy chain variable domain) of an antibody or fragment thereof. Non-limiting examples of antibody variants include single domain antibodies or nanobodies, monospecific Fab 2 , Bispecific Fab 2、 Trispecific Fab 3、 Monovalent IgG, scFv, bispecific antibodies, bispecific diabodies, trispecific triantibodies, scFv-Fc, small antibodies, IgNAR, V-NAR, hcIgG, VhH or peptibody. "Peptibody" as provided herein refers to the peptide portion of the Fc domain of the antibody (by covalent or non-covalent linkers). Other non-limiting examples of antibody variants known in the art include antibodies produced by cartilaginous fish or camelids. The general description of the antibodies and variable regions of camelids and their production, separation and use methods can be found in references WO97 / 49805 and WO 97 / 49805, which are incorporated herein by reference in their entirety, and are used for all purposes. Similarly, the antibodies and variable regions of cartilaginous fish and their production, separation and use methods can be found in WO2005 / 118629, which are incorporated herein by reference in their entirety, and are used for all purposes.

[0083] The terms "CDR L1", "CDR L2" and "CDR L3" provided herein refer to complementarity determining regions (CDR) 1, 2 and 3 of antibody variable light chains (L). In embodiments, variable light chains provided herein include CDR L1, CDR L2 and CDR L3 in the direction from N-terminus to C-terminus. The terms "CDR H1", "CDR H2" and "CDR H3" provided herein refer to complementarity determining regions (CDR) 1, 2 and 3 of antibody variable heavy chains (H). In embodiments, variable heavy chains provided herein include CDR H1, CDR H2 and CDR H3 in the direction from N-terminus to C-terminus. In embodiments, the CDRs of light chains are referred to as CDR1, CDR2 and CDR3 of VL, and the CDRs of heavy chains are referred to as CDR1, CDR2 and CDR3 of VH. See, e.g., tables provided herein.

[0084] As provided herein, the terms "FR L1", "FR L2", "FR L3" and "FR L4" are used according to their common meanings in the art and refer to the framework regions (FR) 1, 2, 3 and 4 of the variable light (L) chain of an antibody. In an embodiment, the variable light chain provided herein includes FR L1, FR L2, FR L3 and FR L4 in the direction from N-terminal to C-terminal. Similarly, the terms "FR H1", "FR H2", "FR H3" and "FR H4" provided herein are used according to their common meanings in the art and refer to the framework regions (FR) 1, 2, 3 and 4 of the variable heavy (H) chain of an antibody. In an embodiment, the variable heavy chain provided herein includes FR H1, FR H2, FR H3 and FR H4 in the direction from N-terminal to C-terminal.

[0085] Exemplary immunoglobulin (antibody) structural units include tetramers. Each tetramer is composed of two pairs of identical polypeptide chains, each pair of polypeptide chains having a "light" (about 25kd) chain and a "heavy" chain (about 50-70kd). The N-terminus of each chain defines a variable region of about 100 to 110 or more amino acids, which is primarily responsible for antigen recognition. The terms variable light chain (VL), variable light chain (VL) domain or light chain variable region and variable heavy chain (VH), variable heavy chain (VH) domain or heavy chain variable region refer to these light chains and heavy chain regions, respectively. The terms variable light chain (VL), variable light chain (VL) domain and light chain variable region referred to herein can be used interchangeably. The terms variable heavy chain (VH), variable heavy chain (VH) domain and heavy chain variable region referred to herein can be used interchangeably. Fc (i.e., fragment crystallizable region) is the "base" or "tail" of an immunoglobulin and is generally composed of two heavy chains that provide two or three constant domains, depending on the type of antibody. By binding to specific proteins, the Fc region ensures that each antibody produces an appropriate immune response to a given antigen. The Fc region also binds to various cell receptors (such as Fc receptors) and other immune molecules (such as complement proteins). The term "light chain" is used according to its common meaning in the biological field and refers to a polypeptide formed by a light chain variable domain (VL) and a light chain constant domain (CL). Similarly, the term "heavy chain" is used according to its common meaning in the biological field and refers to a polypeptide formed by a heavy chain variable domain (VH) and one or more heavy chain constant domains (CH1, CH2, CH3).

[0086] The term "antibody" is used in accordance with its commonly known meaning. Antibodies exist, for example, as intact immunoglobulins or as a number of well-characterized fragments produced by digestion with various peptidases. Thus, for example, pepsin digests an antibody below the disulfide bonds in the hinge region to produce F(ab)' 2 , which is a dimer of Fab, which is itself connected to V by disulfide bonds H -C H1 Attached light chain. F(ab)' 2 The F(ab)' can be reduced under mild conditions to break the disulfide bonds in the hinge region. 2Dimer is converted into Fab' monomer. Fab' monomer is basically Fab with a part of hinge region (see Fundamental Immunology (Paul ed., 3d ed. 1993)). Although various antibody fragments are defined with respect to digestion of intact antibodies, it will be appreciated by those skilled in the art that such fragments can be synthesized de novo chemically or by using recombinant DNA methods. Therefore, as used herein, the term antibody also includes antibody fragments produced by modifying intact antibodies, or those synthesized de novo using recombinant DNA methods (e.g., single-chain Fv) or those identified using phage display libraries (see, e.g., McCafferty et al. Nature (Nature) 348: 552-554 (1990)). The term "antibody" as referred to herein also includes antibody variants, such as single domain antibodies. Therefore, in embodiments, antibodies include single monomer variable antibody domains. Therefore, in embodiments, antibodies include variable light chain (VL) domains or variable heavy chain (VH) domains. In embodiments, antibodies are variable light chain (VL) domains or variable heavy chain (VH) domains.

[0087] To prepare monoclonal or polyclonal antibodies, any technique known in the art may be used (see, e.g., Kohler and Milstein, Nature 256:495-497 (1975); Kozbor et al., Immunology Today 4:72 (1983); Cole et al., Monoclonal Antibodies and Cancer Therapy, pp. 77-96 (1985)). A "monoclonal" antibody (mAb) refers to an antibody derived from a single clone. The technique for producing single-chain antibodies (U.S. Pat. No. 4,946,778) may be adapted to produce antibodies against the polypeptides of the invention. Similarly, transgenic mice or other organisms (such as other mammals) may also be used to express humanized antibodies. Alternatively, phage display technology can be used to identify antibodies and heterologous Fab fragments that specifically bind to a selected antigen (see, e.g., McCafferty et al., Nature 348:552-554 (1990); Marks et al., Biotechnology 10:779-783 (1992)).

[0088] Single chain variable fragment (scFv) is usually a fusion protein of the variable domains of the heavy chain (VH) and light chain (VL) of an immunoglobulin connected by a short linker peptide of 10 to about 25 amino acids. The linker can usually be rich in glycine to improve flexibility, and serine or threonine to improve solubility. The linker can connect the N-terminus of VH to the C-terminus of VL, and vice versa.

[0089] The epitope of mAb is the region where mAb binds to its antigen. If each of the two antibodies competitively inhibits (blocks) the binding of another antibody to the antigen, the two antibodies will bind to the same or overlapping epitopes. That is, as determined in a competitive binding assay, 1x, 5x, 10x, 20x or 100x excess of one antibody inhibits the binding of another antibody by at least 30%, but preferably 50%, 75%, 90% or even 99% (see, e.g., Junghans et al., Cancer Res. 50: 1495, 1990). Alternatively, if substantially all amino acid mutations in the antigen that reduce or eliminate the binding of one antibody reduce or eliminate the binding of another antibody, the two antibodies have the same epitope. If some amino acid mutations that reduce or eliminate the binding of one antibody reduce or eliminate the binding of another antibody, the two antibodies have overlapping epitopes.

[0090] As provided herein, the term "antigen" refers to a molecule that can bind to an antibody binding domain provided herein. As provided herein, an "antigen binding domain" is an antibody region that binds to an antigen (epitope). As described above, an antigen binding domain is generally composed of a constant domain and a variable domain (VL, VH, CL and CH1, respectively) of each heavy chain and light chain. The paratope or antigen binding site is formed at the N-terminus of the antigen binding domain. The two variable domains of the antigen binding domain are generally bound to an epitope on an antigen.

[0091] To prepare suitable antibodies of the invention and uses according to the invention, e.g., recombinant, monoclonal or polyclonal antibodies, many techniques known in the art can be used (see, e.g., Kohler and Milstein, Nature 256:495-497 (1975); Kozbor et al., Immunology Today 4:72 (1983); Cole et al., Monoclonal Antibodies and Cancer Therapy, pp. 77-96, Alan R. Liss, Inc. (1985); Coligan, Current Protocols in Immunology (1991); Harlow & Lane, Antibodies, A Laboratory Manual (1988); and Goding, Monoclonal Antibodies: Principles and Practice (2nd edition, 1986)). Genes encoding the heavy and light chains of the antibody of interest can be cloned from cells, e.g., genes encoding monoclonal antibodies can be cloned from hybridomas, and used to produce recombinant monoclonal antibodies. Gene libraries encoding monoclonal antibody heavy and light chains can also be prepared from hybridomas or plasma cells. The random combination of heavy and light chain gene products produces a large number of antibodies with different antigenic specificities (see, e.g., Kuby, Immunology (3rd edition, 1997)). The technology for producing single-chain antibodies or recombinant antibodies (U.S. Patent No. 4,946,778, U.S. Patent No. 4,816,567) can be suitable for producing antibodies against polypeptides of the present invention. Likewise, transgenic mice or other organisms (such as other mammals) can also be used to express humanized or human antibodies (see, e.g., U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016, Marks et al., Bio / Technology 10:779-783 (1992); Lonberg et al., Nature 368:856-859 (1994); Morrison, Nature 368:812-13 (1994); Fishwild et al., Nature Biotechnology 14:845-51 (1996); Neuberger, Nature Biotechnology 14:826 (1996); and Lonberg & Huszar, Intern. Rev. Immunol.) 13:65-93 (1995).Alternatively, phage display technology can be used to identify antibodies and heterologous Fab fragments that specifically bind to a selected antigen (see, e.g., McCafferty et al., Nature 348:552-554 (1990); Marks et al., Biotechnology 10:779-783 (1992)). Antibodies can also be prepared to be bispecific, i.e., capable of recognizing two different antigens (see, e.g., WO 93 / 08829, Traunecker et al., EMBO J. 10:3655-3659 (1991); and Suresh et al., Methods in Enzymology 121:210 (1986)). Antibodies can also be heterologous conjugates, such as two covalently linked antibodies or immunotoxins (see, e.g., U.S. Pat. No. 4,676,980, WO 91 / 00360; WO 92 / 200373; and EP 03089).

[0092] Methods for humanizing or primatizing non-human antibodies are well known in the art (e.g., U.S. Pat. Nos. 4,816,567; 5,530,101; 5,859,205; 5,585,089; 5,693,761; 5,693,762; 5,777,085; 6,180,370; 6,210,671; and 6,329,511; WO 87 / 02671; European Patent Application 0173494; Jones et al., (1986) Nature 321:522; and Verhoyen et al., (1988) Science 239:1534). Humanized antibodies are further described, for example, in Winter and Milstein (1991) Nature 349:293. Typically, a humanized antibody has one or more amino acid residues introduced into it from a non-human source. These non-human amino acid residues are often referred to as import residues, which are typically taken from an import variable domain. Humanization can essentially be performed according to the method of Winter and colleagues (see, e.g., Morrison et al., PNAS USA, 81:6851-6855 (1984), Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-327 (1988); Morrison and Oi, Adv. Immunol., 44:65-92 (1988), Verhoeyen et al., Science 239:1534-1536 (1988) and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992), Padlan, Molec. Immun., 28:489-498 (1991); Padlan, Molec. 31 (3): 169-217 (1994)), by replacing the corresponding sequence of a human antibody with a rodent CDR or CDR sequence. Thus, such humanized antibodies are chimeric antibodies (US Pat. No. 4,816,567), in which substantially less than the entire human variable domain has been replaced by the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies. For example, a polynucleotide comprising a first sequence encoding a humanized immunoglobulin framework region and a second sequence set encoding a desired immunoglobulin complementarity determining region can be produced synthetically or by combining appropriate cDNA and genomic DNA segments. Human constant region DNA sequences can be isolated from various human cells according to well-known methods.

[0093] "Chimeric antibodies" are antibody molecules in which (a) the constant region or a portion thereof is altered, replaced or exchanged so that the antigen binding site (e.g., the variable region comprising domains VH and VL) is linked to a different or altered class, effector function and / or species, or a completely different molecule that confers new properties to the chimeric antibody, such as a constant region of an enzyme, toxin, hormone, growth factor, drug, etc.; or (b) the variable region or a portion thereof is altered, replaced or exchanged with a variable region having a different or altered antigenic specificity. Preferred antibodies of the invention and for use according to the invention include humanized and / or chimeric monoclonal antibodies.

[0094] When referring to a protein or peptide, the phrase "specifically (or selectively) binds" an antibody or "specifically (or selectively) immunoreacts with..." refers to a binding reaction that determines the presence of a protein, typically in a heterogeneous population of proteins and other biological preparations. Thus, under specified immunoassay conditions, the binding of a specified antibody to a specific protein is at least twice the background, and more typically more than 10 to 100 times the background. Specific binding to an antibody under such conditions requires the selection of an antibody that is specific for a particular protein. For example, polyclonal antibodies can be selected to obtain only a subset of antibodies that specifically immunoreact with a selected antigen and not with other proteins. This selection can be achieved by subtracting antibodies that cross-react with other molecules. A variety of immunoassay formats can be used to select antibodies that specifically immunoreact with a particular protein. For example, solid phase ELISA immunoassays are routinely used to select antibodies that specifically immunoreact with proteins (see, e.g., Harlow and Lane, Using Antibodies, A Laboratory Manual (1998) for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity).

[0095] "Ligand" refers to an agent, such as a polypeptide or other molecule, that is capable of binding to a receptor or an antibody, antibody variant, antibody region or fragment thereof. In an embodiment, the ligand is an antigen, a fragment or portion thereof.

[0096] The techniques for conjugating therapeutic agents to antibodies are well known (see, e.g., Arnon et al., "Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy" in Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985); Hellstrom et al., "Antibodies For Drug Delivery" in Controlled Drug Delivery (2nd ed.), Robinson et al. (eds.), pp. 623-53 (Marcel Dekker, Inc. 1987); Thorpe, "Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review" in Monoclonal Antibodies '84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475-506 (1985); and Thorpe et al., "The Preparation And Cytotoxic Properties Of Antibody-Toxin As used herein, the term "antibody-drug conjugate" or "ADC" refers to a therapeutic agent that is coupled or otherwise covalently bound to an antibody.

[0097] For specific proteins described herein, the protein includes any of a naturally occurring form of the protein, a variant or homologue that maintains protein transcription factor activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to the native protein). In some embodiments, the variant or homologue has at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., 50, 100, 150 or 200 consecutive amino acid portions) compared to the naturally occurring form. In other embodiments, the protein is a protein identified by its NCBI sequence reference. In other embodiments, the protein is a protein identified by its NCBI sequence reference, its homologue or functional fragment.

[0098] As used herein, the term "EGFR protein" or "EGFR" includes a recombinant or naturally occurring form of the epidermal growth factor receptor (also known as proto-oncogene c-ErbB-1, receptor tyrosine protein kinase erbB-1, ERBB, ERBB1, HER1) that maintains EGFR activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to EGFR), or any of its variants or homologs. In some aspects, the variant or homolog has at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity over the entire sequence or a portion of the sequence (e.g., 50, 100, 150 or 200 consecutive amino acid portions) compared to the naturally occurring EGFR protein. In an embodiment, the EGFR protein is substantially identical to the protein identified by UniProt reference number P00533 or a variant or homolog substantially identical thereto.

[0099] Epidermal growth factor receptor, also referred to as EGFR, ErbB1 and HER1, is a cell surface receptor of the epidermal growth factor family of extracellular ligands. The change of EGFR activity is associated with certain cancers. In embodiments, a gene encoding an EGFR polypeptide is provided, which is formed by removing a polypeptide encoding a membrane distal EGF binding domain and a cytoplasmic signaling tail ("truncated EGFR", "tEGFR" or "EGFRt") but retaining a nucleic acid sequence of an extracellular domain IV epitope identified by any anti-EGFR antibody provided herein (e.g., anti-domain IV EGFR antibody, including embodiments thereof). In embodiments, tEGFR does not include EGFR domain III.

[0100] The term "gene" means a segment of DNA involved in producing a protein; it includes the regions before and after the coding region (leader and trailer) and intervening sequences (introns) between the individual coding segments (exons). Leaders, trailers, and introns contain regulatory elements essential for gene transcription and translation. Further, a "protein gene product" is a protein expressed by a specific gene.

[0101] The term "plasmid", "vector" or "expression vector" refers to a nucleic acid molecule encoding a gene and / or regulatory elements necessary for gene expression. Gene expression from a plasmid can occur in cis or in trans. If a gene is expressed in cis, the gene and regulatory elements are encoded by the same plasmid. Trans expression refers to a situation in which a gene and regulatory elements are encoded by separate plasmids.

[0102] The terms "transfection" or "transduction" can be used interchangeably and are defined as the process of introducing a nucleic acid molecule or protein into a cell. Nucleic acids are introduced into cells using non-viral or viral-based methods. The nucleic acid molecule can be a gene sequence encoding a complete protein or a functional portion thereof. Non-viral transfection methods include any suitable transfection method that does not use viral DNA or viral particles as a delivery system for introducing nucleic acid molecules into cells. Exemplary non-viral transfection methods include calcium phosphate transfection, liposome transfection, nucleofection, sonoporation, transfection by heat shock, magnetofection, and electroporation. In some embodiments, electroporation is used to introduce nucleic acid molecules into cells according to standard procedures known in the art. For viral-based transfection methods, any useful viral vector can be used for the methods described herein. Examples of viral vectors include, but are not limited to, retroviral, adenoviral, lentiviral, and adeno-associated viral vectors. In some embodiments, retroviral vectors are used to introduce nucleic acid molecules into cells according to standard procedures known in the art. The term "transfection" or "transduction" also refers to the introduction of proteins into cells from an external environment. Generally, the transduction or transfection of proteins relies on the connection of peptides or proteins that can pass through the cell membrane to related proteins. See, e.g., Ford et al. (2001) Gene Therapy 8: 1-4 and Prochiantz (2007) Nat. Methods 4: 119-20.

[0103] A "label" or "detectable moiety" is a composition detectable by spectroscopic, photochemical, biochemical, immunochemical, chemical or other physical means. For example, useful labels include 32P, fluorescent dyes, electron-dense reagents, enzymes (e.g., enzymes commonly used in ELISA), biotin, digoxigenin or haptens and proteins or other entities that can be detected, such as by incorporating a radioactive label into a peptide or antibody that specifically reacts with the target peptide. Any suitable method known in the art for conjugating an antibody to a label can be used, such as the method described in Hermanson, Bioconjugate Techniques, 1996, Academic Press, Inc., San Diego.

[0104] When the label or detectable moiety is a radioactive metal ion or a paramagnetic ion, the reagent can react with another long tail reactant having a long tail with one or more chelating groups attached to the long tail for binding these ions. The long tail can be a polymer such as polylysine, a polysaccharide or other derivatized or derivatizable chain with side groups to which metals or ions can be added for binding. Examples of chelating groups that can be used according to the present disclosure include, but are not limited to, ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), DOTA, NOTA, NETA, TETA, porphyrins, polyamines, crown ethers, bis-thiosemicarbazones, polyoximes, and the like. Chelates are typically linked to PSMA antibodies or functional antibody fragments via a group that is capable of forming a bond to the molecule with minimal loss of immunoreactivity and minimal aggregation and / or internal crosslinking. When complexed with non-radioactive metals such as manganese, iron, and gadolinium, the same chelates can be used for MRI when used with the antibodies and carriers described herein. Macrocyclic chelates such as NOTA, DOTA and TETA can be used with a variety of metals and radiometals including but not limited to radionuclides of gallium, yttrium and copper, respectively. Other cyclic chelates such as macrocyclic polyethers can be used which are useful for stably binding nuclides such as RAIT. 223 Ra is of great significance. In certain embodiments, the chelating moiety can be used to attach PET imaging agents such as Al- 18 The F complex is linked to a targeting molecule for PET analysis.

[0105] "Contacting" is used according to its plain ordinary meaning and refers to the process of bringing at least two different species (e.g., antibody and antigen) into close enough proximity to react, interact, or physically touch. However, it should be understood that the resulting reaction product may be produced directly from a reaction between the added reagents or from an intermediate from one or more of the added reagents that may be produced in the reaction mixture.

[0106] The term "contacting" can include allowing two species to react, interact, or physically touch, where the two species can be, for example, a pharmaceutical composition provided herein and a cell. In embodiments, contacting includes, for example, allowing a pharmaceutical composition described herein to interact with a cell.

[0107] As used herein, "cell" refers to a cell that performs metabolic or other functions sufficient to maintain or replicate its genomic DNA. Cells can be identified by methods well known in the art, including, for example, the presence of an intact membrane, staining with a specific dye, the ability to reproduce offspring, or, if gametes are present, the ability to combine with a second gamete to produce viable offspring. Cells can include prokaryotic cells and eukaryotic cells. Prokaryotic cells include, but are not limited to, bacteria. Eukaryotic cells include, but are not limited to, yeast cells and cells derived from plants and animals, such as mammals, insects (e.g., moths) and human cells.

[0108] The term "recombinant" when used with reference to, for example, a cell, nucleic acid, protein or vector, means that the cell, nucleic acid, protein or vector has been modified by the introduction of a heterologous nucleic acid or protein or by the alteration of a native nucleic acid or protein, or that the cell is derived from a cell so modified. Thus, for example, a recombinant cell expresses genes not present in the native (non-recombinant) form of the cell or expresses native genes that are otherwise abnormally expressed, underexpressed or not expressed at all. Transgenic cells and plants are those that express heterologous genes or coding sequences, typically as a result of recombinant methods.

[0109] The term "isolated" when applied to a nucleic acid or protein means that the nucleic acid or protein is substantially free of other cellular components with which it is associated in its native state. It may, for example, be in a homogeneous state and may be in a dry state or in aqueous solution. Purity and homogeneity are typically determined using analytical chemistry techniques such as polyacrylamide gel electrophoresis or high performance liquid chromatography. The major protein present in the preparation is substantially purified.

[0110] The term "heterologous" when used with reference to portions of a nucleic acid indicates that the nucleic acid comprises two or more subsequences that are not found in the same relationship to each other in nature. For example, nucleic acids are often recombinantly produced, having two or more sequences from unrelated genes arranged to produce a new functional nucleic acid, such as a promoter from one source and a coding region from another source. Similarly, a heterologous protein indicates that the protein comprises two or more subsequences that are not found in the same relationship to each other in nature (e.g., a fusion protein).

[0111] The term "exogenous" refers to a molecule or substance (e.g., a compound, nucleic acid, or protein) that originates from outside a given cell or organism. For example, an "exogenous promoter" as referred to herein is a promoter that does not originate from the cell or organism in which it is expressed. In contrast, the term "endogenous" or "endogenous promoter" refers to a molecule or substance that is native to a given cell or organism or that originates from within a given cell or organism.

[0112] The term "expression" includes any step involved in producing a polypeptide, including but not limited to transcription, post-transcriptional modification, translation, post-translational modification and secretion. Expression can be detected using conventional techniques for detecting protein (e.g., ELISA, Western blotting, flow cytometry, immunofluorescence, immunohistochemistry, etc.).

[0113] "Biological sample" or "sample" refers to material obtained from or derived from a subject or patient. Biological samples include tissue sections, such as biopsy and autopsy samples, and frozen sections for histological purposes. Such samples include body fluids, such as blood and blood parts or products (e.g., serum, plasma, platelets, red blood cells, etc.), sputum, tissues, cultured cells (e.g., primary cultures, explants, and transformed cells), feces, urine, synovial fluid, joint tissues, immune cells, hematopoietic cells, fibroblasts, macrophages, T cells, etc. Biological samples are typically obtained from eukaryotic organisms, such as mammals, such as primates, e.g., chimpanzees or humans; cattle; dogs; cats; rodents, e.g., guinea pigs, rats, mice; rabbits; or birds; reptiles; or fish.

[0114] "Control" or "standard control" refers to a sample, measurement or value used as a reference, usually a known reference, for comparison with a test sample, measurement or value. For example, a test sample can be collected from a patient suspected of having a given disease (e.g., cancer) and compared with a known normal (non-disease) individual (e.g., a standard control subject). The standard control can also represent a population of similar individuals (e.g., standard control subjects) that have never suffered from a given disease (i.e., a standard control population), for example, an average measurement or value collected from healthy individuals with similar medical backgrounds, the same age, weight, etc. The standard control value can also be obtained from the same individual, such as an early sample obtained from a patient before the onset of the disease. For example, a control can be designed to compare the therapeutic benefits based on pharmacological data (e.g., half-life) or therapeutic measures (e.g., comparison of side effects). Controls are also valuable for determining the significance of data. For example, if the value of a given parameter varies widely in the control, the variation in the test sample will not be considered significant. The technician will recognize that standard controls can be designed to evaluate any number of parameters (e.g., RNA levels, protein levels, specific cell types, specific body fluids, specific tissues, etc.).

[0115] Those skilled in the art will understand which standard control is most appropriate in a given situation and can analyze data based on comparisons with standard control values. Standard controls are also valuable for determining the significance (e.g., statistical significance) of data. For example, if the values ​​of a given parameter in the standard control vary widely, then the changes in the test samples will not be considered significant.

[0116] "Patient" or "subject in need thereof" refers to a living organism suffering from or susceptible to a disease or condition that can be treated by administering a composition or pharmaceutical composition as provided herein. Non-limiting examples include humans, other mammals, cattle, rats, mice, dogs, monkeys, goats, sheep, cows, deer, and other non-mammals. In some embodiments, the patient is a human.

[0117] The term "disease" or "condition" refers to a state or health status of a patient or subject that can be treated with the compounds or methods provided herein. The disease can be cancer. Cancer can refer to a malignant solid tumor. Malignant solid tumors include malignant tumors that may not have fluid or cysts. For example, malignant solid tumors can include breast cancer, ovarian cancer, pancreatic cancer, cervical cancer, gastric cancer, kidney cancer, head and neck cancer, bone cancer, skin cancer, or prostate cancer. In some additional instances, "cancer" refers to human cancer and carcinomas, sarcomas, adenocarcinomas, lymphomas, leukemias, including solid and lymphomas, renal cancer, breast cancer, lung cancer, bladder cancer, colon cancer, ovarian cancer, prostate cancer, pancreatic cancer, stomach cancer, brain cancer, head and neck cancer, skin cancer, uterine cancer, testicular cancer, gliomas, esophageal cancer, and liver cancer (including hepatocarcinomas), lymphomas (including acute B lymphoblastic lymphomas), non-Hodgkin's lymphomas (e.g., Burkitt's lymphoma, small cell lymphoma, and large cell lymphoma), Hodgkin's lymphoma, leukemias (including acute myeloid leukemia (AML), ALL, and CML), or multiple myeloma.

[0118] As used herein, the term "cancer" refers to all types of cancers, neoplasms or malignancies found in mammals (e.g., humans), including leukemias, carcinomas and sarcomas. Exemplary cancers that can be treated with the compounds or methods provided herein include breast cancer, colon cancer, kidney cancer, leukemia, lung cancer, melanoma, ovarian cancer.

[0119] The term "modulate" is used according to its ordinary general meaning and refers to the act of changing or altering one or more properties. "Modulate" refers to the process of changing or varying one or more properties. For example, when applied to the effect of a modulator on a target protein, modulating means changing by increasing or decreasing a property or function of the target molecule or the amount of the target molecule.

[0120] In the case where a substance or substance activity or function is associated with a disease (e.g., a protein-related disease, a cancer (e.g., breast cancer, lung cancer)), the term "associated" or "associated with..." means that the disease (e.g., cancer) is (completely or partially) caused by the substance or substance activity or function, or that the symptoms of the disease are (completely or partially) caused by the substance or substance activity or function. As used herein, a pathogen is described as being associated with a disease if it can be a target for treating the disease.

[0121] As used herein, the term "abnormal" means different from normal. When used to describe enzymatic activity, abnormal refers to an activity that is greater or less than the average of a normal control or normal non-disease control sample. Abnormal activity can refer to an amount of activity that causes a disease, wherein returning the abnormal activity to a normal or non-disease associated amount (e.g., by using a method as described herein) results in a reduction in the disease or one or more disease symptoms.

[0122] As referred to herein, a "therapeutic agent" is a composition used to treat or prevent diseases such as cancer (e.g., leukemia). In embodiments, the therapeutic agent is an anticancer agent. "Anticancers" are used in accordance with their simple common meaning and refer to compositions (e.g., compounds, drugs, antagonists, inhibitors, regulators) that have antitumor properties or the ability to inhibit cell growth or proliferation. In embodiments, anticancer agents are chemotherapeutics. In embodiments, anticancer agents are reagents identified herein that have utility in methods for treating cancer. In embodiments, anticancer agents are reagents approved for the treatment of cancer by similar regulatory agencies in countries outside the FDA or the United States.

[0123] As used herein, "treating" a condition, disease or disorder or a symptom associated with a condition, disease or disorder refers to a method for obtaining a beneficial or desired result, including a clinical result. Beneficial or desired clinical results may include, but are not limited to, alleviating or improving one or more symptoms or conditions, alleviating the degree of a condition, disorder or disease, stabilizing the state of a condition, disorder or disease, preventing a condition, disorder or disease, preventing the spread of a condition, disorder or disease, delaying or slowing down a condition, disorder or disease progression, delaying or slowing down a condition, disorder or disease onset, improving or alleviating a condition, disorder or disease state, and alleviating, whether in part or in whole. "Treatment" may also mean extending the survival time of a subject beyond the expected survival time when no treatment is performed. "Treatment" may also mean inhibiting the progress of a condition, disorder or disease, temporarily slowing down the progress of a condition, disorder or disease, although in some cases, it involves permanently stopping the progress of a condition, disorder or disease. As used herein, the term "treatment" refers to a method for reducing the effect of one or more symptoms of a disease or condition characterized by protease expression or a symptom of a disease or condition characterized by protease expression. Thus, in the disclosed methods, treatment may refer to a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% reduction in severity of a determined disease, condition, or symptom of a disease or condition. For example, if one or more disease symptoms in a subject are reduced by 10% compared to a control, a method of treating a disease is considered treatment. Thus, a reduction may be 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any percentage reduction between 10% and 100% compared to a natural or control level. It should be understood that treatment does not necessarily refer to a cure or complete ablation of a disease, condition, or symptom of a disease or condition. Furthermore, as used herein, references to reduction, decrease or inhibition include changes of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater compared to control levels, and such terms may include but do not necessarily include complete ablation.

[0124] The term "dose" is used interchangeably herein. Dose refers to the amount of active ingredient given to an individual at each administration. The dose will vary according to a variety of factors, including the normal dose range for a given therapy, the frequency of administration; the size and tolerance of the individual; the severity of the condition; the risk of side effects; and the route of administration. Those skilled in the art will recognize that the dose can be modified based on the above factors or based on the progress of the treatment. The term "dosage form" refers to the specific form of a drug or pharmaceutical composition and depends on the route of administration. For example, the dosage form can be in liquid form for atomization, such as for inhalation, in tablet or liquid form, such as for oral delivery, or in saline solution, such as for injection.

[0125] A "control" sample or value refers to a sample used as a reference, typically a known reference, to be compared with a test sample. For example, a test sample can be collected from a test condition, such as in the presence of a test compound, and compared with a sample under known conditions, such as in the absence of a test compound (negative control), or in the presence of a known compound (positive control). A control can also represent a mean value collected from multiple tests or results. Those skilled in the art will recognize that controls can be designed to assess any number of parameters. For example, controls can be designed to compare therapeutic benefits based on pharmacological data (e.g., half-life) or therapeutic measures (e.g., comparison of side effects). Those skilled in the art will understand which controls are valuable in a given situation, and can analyze data based on comparisons with control values. Controls are also valuable for determining the significance of data. For example, if the value of a given parameter varies widely in a control, the variation in the test sample will not be considered significant.

[0126] "Patient" or "subject in need thereof" refers to a living organism suffering from or susceptible to a disease or condition that can be treated by administering a composition or pharmaceutical composition as provided herein. Non-limiting examples include humans, other mammals, cattle, rats, mice, dogs, monkeys, goats, sheep, cows, deer, and other non-mammals. In some embodiments, the patient is a human.

[0127] The term "disease" or "condition" refers to a state or health condition of a patient or subject that can be treated with a compound, pharmaceutical composition, or method as provided herein. In an embodiment, the disease is cancer (e.g., lung cancer, ovarian cancer, osteosarcoma, bladder cancer, cervical cancer, liver cancer, kidney cancer, skin cancer (e.g., Merkel cell carcinoma), testicular cancer, leukemia, lymphoma (Mantel cell lymphoma), head and neck cancer, colorectal cancer, prostate cancer, pancreatic cancer, melanoma, breast cancer, neuroblastoma).

[0128] As used herein, the term "cancer" refers to all types of cancers, tumors or malignancies found in mammals, including leukemias, lymphomas, melanomas, neuroendocrine tumors, carcinomas and sarcomas. Exemplary cancers that can be treated with the compounds, pharmaceutical compositions or methods provided herein include lymphomas (e.g., Mantel cell lymphomas, follicular lymphomas, diffuse large B-cell lymphomas, marginal band lymphomas, Burkitt's lymphomas), sarcomas, bladder cancer, bone cancer, brain tumors, cervical cancer, colon cancer, esophageal cancer, gastric cancer, head and neck cancer, kidney cancer, myeloma, thyroid cancer, leukemia, prostate cancer, breast cancer (e.g., triple negative, ER positive, ER negative, chemotherapy resistance, Herceptin resistance, HER2 positive, doxorubicin resistance, tamoxifen resistance, ductal carcinoma, lobular carcinoma, primary, metastatic) , ovarian cancer, pancreatic cancer, liver cancer (e.g., hepatocellular carcinoma), lung cancer (e.g., non-small cell lung cancer, squamous cell lung cancer, adenocarcinoma, large cell lung cancer, small cell lung cancer, carcinoid, sarcoma), glioblastoma multiforme, glioma, melanoma, prostate cancer, castration-resistant prostate cancer, breast cancer, triple-negative breast cancer, glioblastoma, ovarian cancer, lung cancer, squamous cell carcinoma (e.g., head, neck or esophageal), colorectal cancer, leukemia (e.g., lymphocytic leukemia, chronic lymphocytic leukemia, hairy cell leukemia), acute myeloid leukemia, lymphoma, B-cell lymphoma or multiple myeloma. Additional examples include thyroid cancer, cancer of the endocrine system, brain cancer, breast cancer, cervical cancer, colon cancer, head and neck cancer, esophageal cancer, liver cancer, kidney cancer, lung cancer, non-small cell lung cancer, melanoma, mesothelioma, ovarian cancer, sarcoma, stomach cancer, uterine cancer, or medulloblastoma, Hodgkin lymphoma, non-Hodgkin lymphoma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, ovarian cancer, rhabdomyosarcoma, essential thrombocythaemia, essential macroglobulinemia, primary brain tumor, cancer, malignant islet cell tumor, malignant carcinoid, bladder cancer, precancerous skin lesions, testicular cancer, lymphoma, thyroid cancer, neuroblastoma, esophageal cancer, genitourinary tract cancer, malignant hypercalcemia, endometrial cancer, adrenocortical carcinoma, endocrine or exocrine pancreatic tumor, medullary thyroid cancer, medullary thyroid cancer, melanoma, colorectal cancer, papillary thyroid carcinoma, hepatocellular carcinoma, Paget's disease of the nipple, phyllodes tumors, lobular carcinoma, ductal carcinoma, pancreatic stellate cell carcinoma, hepatic stellate cell carcinoma, or prostate cancer.

[0129] The term "leukemia" refers to a progressive, malignant disease of the blood-forming organs and is generally characterized by the dysregulated proliferation and development of leukocytes and their precursors in the blood and bone marrow. The clinical classification of leukemias is generally based on (1) the duration and characteristics of the disease - acute or chronic; (2) the cell types involved; myeloid (myelocytic), lymphoid (lymphoblastic), or monocytic; (3) an increase or absence of an increase in the number of abnormal cells in the leukemia or non-leukemia (subleukemia). The P388 leukemia model is widely accepted as a predictor of in vivo anti-leukemia activity. It is believed that compounds that test positive in the P388 assay will generally exhibit a certain level of anti-leukemia activity (in vivo), regardless of the type of leukemia being treated. Thus, the present application includes methods of treating leukemia, and preferably methods of treating the following diseases: acute non-lymphocytic leukemia, chronic lymphocytic leukemia, acute granulocytic leukemia, chronic granulocytic leukemia, acute promyelocytic leukemia, adult T-cell leukemia, leukopenic leukemia, leukemic leukemia, basophilic leukemia, blast cell leukemia, bovine leukemia, chronic myeloid leukemia, cutaneous leukemia, embryonic leukemia, eosinophilic leukemia, Gross' leukemia, hairy cell leukemia, hemoblastic leukemia disease (hemoblasticleukemia), hemoblastic leukemia, histiocytic leukemia, stem cell leukemia, acute monocytic leukemia, leukopenic leukemia, lymphocytic leukemia, lymphoblastic leukemia, lymphocytic leukemia, lymphoid leukemia, lymphosarcoma cell leukemia, mast cell leukemia, megakaryocytic leukemia, micromyeloblastic leukemia, monocytic leukemia, myeloblastic leukemia, myelocytic leukemia, myelogranulocytic leukemia, myelomonocytic leukemia, Naegeli's leukemia (Naegeli leukemia), plasma cell leukemia, multiple myeloma, plasma cell leukemia, promyelocytic leukemia, Rieder cell leukemia, Schilling's leukemia, stem cell leukemia, subleukemic leukemia, and undifferentiated cell leukemia.

[0130] As used herein, the terms "metastasis", "metastatic" and "metastatic cancer" can be used interchangeably and refer to proliferative diseases or conditions, such as the spread of cancer from one organ or another non-adjacent organ or body part. Cancer occurs at the starting site (e.g., breast), which is called a primary tumor, such as primary breast cancer. Some cancer cells in the primary tumor or site of origin acquire the ability to penetrate and infiltrate normal tissues around the local area, and / or penetrate the walls of the lymphatic system or the vascular system through systemic circulation to other parts and tissues in the body. The second clinically detectable tumor formed by the cancer cells of the primary tumor is called a metastatic or secondary tumor. When cancer cells metastasize, it is speculated that the metastatic tumor and its cells are similar to the primary tumor. Therefore, if lung cancer metastasizes to the breast, the secondary tumor at the breast site is composed of abnormal lung cells rather than abnormal breast cells. Secondary tumors in the breast refer to metastatic lung cancer. Therefore, the phrase metastatic cancer refers to a disease in which a subject suffers from or has suffered from a primary tumor and suffers from one or more secondary tumors. The phrase non-metastatic cancer or a subject with non-metastatic cancer refers to a disease in which a subject has a primary tumor but does not have one or more secondary tumors. For example, metastatic lung cancer refers to a disease in a subject with a primary lung tumor or a history of a primary lung tumor and one or more secondary tumors at a second location or locations (e.g., in the breast).

[0131] In the context of a substance or substance activity or function associated with a disease (e.g., cancer), the term "associated with" or "associated with..." means that the disease is caused (in whole or in part), or the symptoms of the disease are caused (in whole or in part) by the substance or substance activity or function.

[0132] As used herein, "treat" or "alleviate" or "improve" are used interchangeably herein. These terms refer to methods of obtaining beneficial or desired results, including but not limited to therapeutic benefit and / or preventive benefit. By therapeutic benefit is meant the elimination or amelioration of the underlying condition being treated. Moreover, a therapeutic benefit is obtained by the elimination or amelioration of one or more physiological symptoms associated with the underlying condition, such that an improvement is observed in the patient, although the patient may still suffer from the underlying condition. For preventive benefit, the composition can be administered to a patient at risk for developing a particular disease, or to a patient reporting one or more physiological symptoms of a disease, even though a diagnosis of the disease may not have been made. Treatment includes preventing the disease, i.e., preventing the development of clinical symptoms of the disease by administering a protective composition prior to the induction of the disease; inhibiting the disease, i.e., preventing the development of clinical symptoms of the disease by administering a protective composition after the induction event but before the clinical manifestation or re-manifestation of the disease; inhibiting the disease, i.e., preventing the development of clinical symptoms by administering a protective composition after the initial appearance of clinical symptoms; preventing the recurrence of the disease and / or alleviating the disease, i.e., causing the regression of clinical symptoms by administering a protective composition after the initial appearance of clinical symptoms. For example, certain methods herein treat cancer (e.g., lung cancer, ovarian cancer, osteosarcoma, bladder cancer, cervical cancer, liver cancer, kidney cancer, skin cancer (e.g., Merkel cell carcinoma), testicular cancer, leukemia, lymphoma, head and neck cancer, colorectal cancer, prostate cancer, pancreatic cancer, melanoma, breast cancer, neuroblastoma). For example, certain methods herein treat cancer by reducing or decreasing or preventing the occurrence, growth, metastasis or progression of cancer; or by alleviating the symptoms of cancer. Symptoms of cancer (e.g., lung cancer, ovarian cancer, osteosarcoma, bladder cancer, cervical cancer, liver cancer, kidney cancer, skin cancer (e.g., Merkel cell carcinoma), testicular cancer, leukemia, lymphoma, head and neck cancer, colorectal cancer, prostate cancer, pancreatic cancer, melanoma, breast cancer, neuroblastoma) will be known to or can be determined by one of ordinary skill in the art.

[0133] As used herein, the term "treatment", "treatment" or "treatment" refers to a method for reducing the effect of one or more symptoms of a disease or condition characterized by protease expression or a symptom of a disease or condition characterized by protease expression. Therefore, in the disclosed method, treatment can refer to a reduction in the severity of a determined disease, condition or symptom of a disease or condition by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%. For example, if one or more disease symptoms in a subject are reduced by 10% compared to a control, the method of treating the disease is considered to be treatment. Therefore, compared to natural or control levels, reduction can be 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any reduction percentage between 10% and 100%. It should be understood that treatment does not necessarily refer to the cure or complete ablation of a disease, condition or symptom of a disease or condition. Furthermore, as used herein, references to reduction, decrease or inhibition include changes of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater compared to control levels, and such terms may include but do not necessarily include complete ablation.

[0134] The terms "dose" and "dosage" are used interchangeably herein. Dose refers to the amount of active ingredient given to an individual at each administration. The dose will vary depending on a variety of factors, including the normal dose range for a given therapy, the frequency of administration; the size and tolerance of the individual; the severity of the condition; the risk of side effects; and the route of administration. Those skilled in the art will recognize that the dose can be modified based on the above factors or based on the progress of the treatment. The term "dosage form" refers to the specific form of a drug or pharmaceutical composition and depends on the route of administration. For example, the dosage form can be in liquid form for aerosolization, such as for inhalation, in tablet or liquid form, such as for oral delivery, or in saline solution, such as for injection.

[0135] As used herein, "therapeutically effective dose or amount" refers to a dose that produces the effect of its administration (e.g., treatment or prevention of disease). The exact dosage and formulation will depend on the purpose of the treatment and will be determined by those skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (Volumes 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Remington: The Science and Practice of Pharmacy, 20th edition, Gennaro ed. (2003), and Pickar, Dosage Calculations (1999)). For example, for a given parameter, a therapeutically effective amount will show an increase or decrease of at least 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90%, or at least 100%. Therapeutic efficacy can also be expressed as a "fold" increase or decrease. For example, a therapeutically effective amount can have at least 1.2-fold, 1.5-fold, 2-fold, 5-fold or more effects relative to a control. A therapeutically effective dose or amount can improve one or more symptoms of a disease. When the effect of administration is to treat a person at risk of disease, a therapeutically effective dose or amount can prevent or delay the onset of a disease or one or more symptoms of a disease.

[0136] "Effective amount" is an amount sufficient to achieve the purpose (e.g., to achieve its administration effect, treat a disease, reduce enzyme activity, alleviate one or more symptoms of a disease or condition). An example of an "effective amount" is an amount sufficient to help treat, prevent or alleviate one or more symptoms of a disease, which may also be referred to as a "therapeutically effective amount". "Relief" of one or more symptoms (and the grammatical equivalents of this phrase) means reducing the severity or frequency of one or more symptoms, or eliminating one or more symptoms. The "preventive effective amount" of a drug is the amount of the drug with the expected preventive effect when applied to a subject, such as preventing or delaying the onset (or recurrence) of damage, disease, pathology or condition or reducing the possibility of the onset (or recurrence) of damage, disease, pathology or condition or its symptoms. The complete preventive effect does not necessarily occur by administering a single dose, and may occur after only a series of doses are administered. Therefore, the preventive effective amount may be administered in one or more administration forms. As used herein, "activity reduction amount" refers to the amount of antagonist required to reduce the activity of an enzyme or protein relative to the absence of an antagonist. As used herein, "function-destroying amount" refers to the amount of antagonist required to destroy the function of an enzyme or protein relative to the absence of an antagonist. Guidance on appropriate dosages for a given class of drug products can be found in the literature. For example, for a given parameter, an effective amount will show an increase or decrease of at least 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90% or at least 100%. Efficacy can also be expressed as a "-fold" increase or decrease. For example, a therapeutically effective amount can have an effect of at least 1.2 times, 1.5 times, 2 times, 5 times or more relative to a control. The exact amount will depend on the purpose of the treatment, and will be determined by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (Vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th edition, 2003, Gennaro, ed., Lippincott, Williams & Wilkins).

[0137] As used herein, the term "administration" means oral administration to a subject, as a suppository, topical contact, intravenous, intraperitoneal, intramuscular, intralesional, intrathecal, intranasal or subcutaneous administration, or the implantation of a sustained-release device, such as the implantation of a miniature osmotic pump. Administered by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palate, gum, nasal, vaginal, rectal or transdermal). Parenteral administration includes, for example, intravenous, intramuscular, intraarterial, intradermal, subcutaneous, peritoneal, intraventricular and intracranial administration. Other modes of delivery include, but are not limited to, the use of liposome preparations, intravenous infusions, transdermal patches, etc. "Combined administration" refers to the administration of the composition described herein at the same time, at the beginning or at the end of one or more additional therapies, such as cancer therapies, such as chemotherapy, hormonal therapy, radiotherapy or immunotherapy. The compounds of the present invention may be administered alone or may be co-administered to a patient. Co-administration means the administration of compounds (more than one compound) alone or in combination simultaneously or sequentially. Therefore, when necessary, the preparation may also be used in combination with other active substances (e.g., to reduce metabolic degradation). The compositions of the present invention may be delivered via transdermal, topical routes and may be formulated as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, paints, powders and aerosols.

[0138] In embodiments, the methods provided herein further comprise administering to the subject an additional therapeutic agent. As described above, a therapeutic agent is a composition useful for treating or preventing a disease such as cancer. In embodiments, the additional therapeutic agent is an anticancer agent.

[0139] The term "anticancer agent" is used according to its ordinary common meaning and refers to a composition (e.g., compound, drug, antagonist, inhibitor, modulator) having anti-tumor properties or the ability to inhibit cell growth or proliferation. In some embodiments, the anticancer agent is a chemotherapeutic agent. In some embodiments, the anticancer agent is an agent identified herein that has utility in a method for treating cancer. In some embodiments, the anticancer agent is an agent approved by the FDA or similar regulatory agencies in countries other than the United States for the treatment of cancer. Examples of anticancer agents include, but are not limited to, MEK (e.g., MEK1, MEK2, or MEK1 and MEK2) inhibitors (e.g., XL518, CI-1040, PD035901, selumetinib / AZD6244, GSK1120212 / trametinib, GDC-0973, ARRY-162, ARRY-300, AZD8330, PD0325901, U0126, PD98059, TAK-733, PD318088, AS703026, BAY869766), 】 (e.g., cyclophosphamide, ifosfamide, chlorambucil, busulfan, melphalan, mechlorethamine, uramustine, thiotepa, nitrosoureas, nitrogen mustards (e.g., mechlorethamine, cyclophosphamide, chlorambucil, meiphalan), ethyleneimine and methylmelamine (e.g., hexamethylmelamine, thiotepa), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine, lomustine, semustine, streptozotocin), triazenes (dacarbazine decarbazine), antimetabolites (e.g., 5-thioazaprine, leucovorin, capecitabine, fludarabine, gemcitabine, pemetrexed, raltitrexed, folic acid analogs (e.g., methotrexate) or pyrimidine analogs (e.g., fluorouracil, floxuridine, cytarabine), purine analogs (e.g., mercaptopurine, thioguanine, pentostatin, etc.), plant alkaloids (e.g., vincristine, vinblastine, vinorelbine, vindesine, podophyllotoxin, paclitaxel, docetaxel, etc.) , topoisomerase inhibitors (e.g., irinotecan, topotecan, amsacrine, etoposide (VP16), etoposide phosphate, teniposide, etc.), antitumor antibiotics (e.g., doxorubicin, adriamycin, daunorubicin, epirubicin, actinomycin, bleomycin, mitomycin, mitoxantrone, plicamycin, etc.), platinum-based compounds (e.g., cisplatin, oxaloplatin, carboplatin), anthracenediones (e.g., mitoxantrone), substituted ureas (e.g., hydroxyurea), methylhydrazine derivatives Biologics (e.g., procarbazine), adrenocortical suppressants (e.g., mitotane, aminoglutethimide), epipodophyllotoxins (e.g., etoposide), antibiotics (e.g., daunorubicin, doxorubicin, bleomycin), enzymes (e.g., L-asparaginase), mitogen-activated protein kinase signaling inhibitors (e.g., U0126, PD98059, PD184352, PD0325901, ARRY-142886, SB239063, SP600125, BAY 43-9006, wortmannin or LY294002, Syk inhibitors, mTOR inhibitors, antibodies (e.g., rituxan), gossypol, genasense, polyphenol E, Chlorofusin, all-trans retinoic acid (ATRA), bryostatin, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), 5-aza-2'-deoxycytidine, all-trans retinoic acid, doxorubicin, vincristine, etoposide, gemcitabine, imatinib (Gleevec.RTM.), geldanamycin, 17-N-allylamino-17-demethoxygeldanamycin (17-AAG), flavopiridol, LY294002, bortezomib, trastuzumab, BAY11-7082, PKC412, PD184352, 20-epi-1,25-dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonist; hexamethylmelamine; ambamustine; dichlorophenoxyacetic acid ( amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole; andrographolide; angiogenesis inhibitors; antagonist D; antagonist G; antarelix; anti-dorsal morphogenetic protein-1; antiandrogen, prostate cancer; antiestrogens; antineoplastons; antisense oligonucleotides; aphidicolin glycinate; apoptosis gene regulators; apoptosis regulators; apurinic nucleic acids; ara-CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane; atrimustine; axinstatin 1; axinstatin 2; axinstatin 3; azasetron; azatoxin; azatyrosine; baccatin III derivatives; balanol; batimastat; BCR / ABL antagonists; benzochlorins; benzoylstaurosporine; beta-lactam derivatives; beta-alethine; beta-betamycin B; betulinic acid acid; bFGF inhibitors; bicalutamide; bisantrene; bisaziridinylspermine; bisnafide; bistratene AA); bizelesin; breflate; bropirimine; budotitane; buthionine sulfoximine; calcipotriol; calphostin C; camptothecin derivatives; canarypox IL-2; capecitabine; carboxyamidotriazole; carboxyamidotriazole; CaRest M3; CARN 700; chondroitin inhibitor; carzelesin; casein kinase inhibitor (ICOS); castanospermine; cecropin B; cetrorelix; chlorins; chloroquinoxaline sulfonamide sulfonamide; cicaprost; cis-porphyrin; cladribine; clomifene analogues; clotrimazole; collismycin A; collismycin B; combretastatin A4; combretastatin analogues; conagenin; crambescidin 816; crisnatol; cryptophycin 8; cryptophycin A derivatives; curacin A; cyclopentanthraquinones; cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor; cytostatin; dacliximab; decitabine; dehydrodidemnin BB); deslorelin; dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone; didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine; 9-dioxamycin; diphenyl spiromustine; docosanol; dolasetron; doxifluridine; droloxifene; dronabinol; duocarmycin SA SA; ebselen; ecomustine; edelfosine; edrecolomab; eflornithine; elemene; emitefur; epirubicin; epristeride; estramustine analogs analogue; estrogen agonist; estrogen antagonist; etanidazole; etoposide phosphate; exemestane; fadrozole; fazarabine; fenretinide; filgrastim; finasteride; flavopiridol; flezellastine; fluasterone; fludarabine; fluorodaunorunicin hydrochloride hydrochloride; forfenimex; formestane; fostriecin; fotemustine; gadoliniumtexaphyrin; gallium nitrate; galocitabine; ganirelix; gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam; heregulin; hexamethylenebisacetamide; hypericin; ibandronic acidacid; idarubicin; idoxifene; idramantone; ilmofosine; ilomastat; imidazoacridones; imiquimod; immunostimulatory peptides; insulin-like growth factor-1 receptor inhibitors; interferon agonists; interferons; interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact; irsogladine; isobengazole; isohomohalicondrin B; itasetron; jasplakinolide; kahalalide F F); lamellarin-N triacetate; lanreotide; leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole; leukemia inhibitory factor; interferon-alpha; leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole; linear polyamine analogs; lipophilic disaccharide peptides; lipophilic platinum compounds; lissoclinamide 7 7); lobaplatin; lombricine; lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine; lurtotecan; lutetiumtexaphyrin; lysofylline; cleaved peptide; maitansine; mannostatin AA); marimastat; masoprocol; maspin; matrix dissolving factor inhibitor; matrix metalloproteinase inhibitor; menogaril; merbarone; meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine; mirimostim; mismatch double-stranded RNA; mitoguazone; mitolactol; mitomycin analogs; mitonafide; mitoxantrone; mofarotene; molgramostim; monoclonal antibodies, human chorionic gonadotropin; monophosphoryl lipid A+ mycobacterial cell wall sk; mopidamol; multidrug resistance gene inhibitor; multitumor inhibitor 1-based therapy; nitrogen mustard anticancer agent; Indian Ocean sponge B (mycaperoxide B); mycobacterial cell wall extract; myriaporone; N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip; naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin; nemorubicin; neridronic acidacid; neutral endopeptidase; nilutamide; nisamycin; nitric oxide modulators; oxynitrogen antioxidants; nitrullyn; O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone; ondansetron; ondansetron; oracin; oral cytokine inducers; ormaplatin; osaterone; oxaliplatin; oxaunomycin; palauamine; palmitoylrhizoxin; pamidronic acid acid; panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin; pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin; phenylacetate; phosphatase inhibitors; picibanil; pilocarpine hydrochloride; pirarubicin; piritrexim; placetin A A); placetin B; plasminogen activator inhibitors; platinum complexes; platinum compounds; platinum-triamine complexes; porfimersodium; porfiromycin; prednisone; propyl diacridone; prostaglandin J2; proteasome inhibitors; protein A-based immunomodulators; protein kinase C inhibitors; protein kinase C inhibitors, microalgae; protein tyrosine phosphatase inhibitors; purine nucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine; pyridoxal hemoglobin polyoxyethylene conjugates; raf antagonists; raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors; ras inhibitors; ras-GAP inhibitors; retelliptine demethylated; rhenium etidronate Rerhenium Re 186 etidronate; rhizoxin; ribozyme; RII retinamide; rogletimide; rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; safingol; saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetic; semustine; senescence-derived inhibitor 1; sense oligonucleotide; signal transduction inhibitor; signal transduction modulator; single-chain antigen binding protein; sizofuran; sobuzoxane; sodium borocaptate; sodium phenylacetate; solverol; somatomedin binding protein; sonermin; sparsosic acid; spicamycin D; spiromustine; splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem cell division inhibitor; stipiamide; stromelysin inhibitor; sulfinosine; superactive vasoactive intestinal peptide antagonist; suradista; suramin; swainsonine; synthetic glycosaminoglycans; tallimustine; tamoxifen methyl iodide; tauromustine; tazarotene; tecogalan sodium sodium; tegafur; tellurapyrylium; telomerase inhibitors; temoporfin; temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine; thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetics; thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroid stimulating hormone; tinethylprotoporphyrinethyletiopurpurin; tirapazamine; titanocenyl dichloride; topsentin; toremifene; totipotent stem cell factor; translation inhibitors; tretinoin; triacetyluridine; triciribine; trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenital sinus-derived growth inhibitory factor; urokinase receptor antagonists; vapreotide; variolin B B); vector system, red blood cell gene therapy; velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine; vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; zinostatin stimalamer, doxorubicin, Dactinomycin, bleomycin, vinblastine, cisplatin, acivicin, aclarubicin, acodazole hydrochloride, acronin, adozelesin, aldesleukin, hexamethylmelamine, ambomycin, ametantron acetate acetate; aminoglutethimide; amsacrine; anastrozole; anthramycin; asparaginase; asperlin; azacitidine; azetepa; azotomycin; batimastat; benzodepa; bicalutamide; bisantrene hydrochloride; binasnafide dimesylate; bizelesin; bleomycin sulfatesulfate; brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone; caracemide; carbetimer; carboplatin; carmustine; carubicin hydrochloride; carzelesin; cedefingol; chlorambucil; cirolemycin; cladribine; crisnatol mesylate; cyclophosphamide; cytarabine; dacarbazine; daunorubicin hydrochloride hydrochloride; decitabine; dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone; doxorubicin; doxorubicin hydrochloride; droloxifene; droloxifene citrate; dromostanolone propionate; duazomycin; edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatin; enpromate; epipropidine; epirubicin hydrochloride hydrochloride; erbulozole; esorubicin hydrochloride; estramustine; estramustine phosphatesodium; etanidazole; etoposide; etoposide phosphate; etoprine; fadrozole hydrochloride; fazarabine; fenretinide; floxuridine; fludarabine phosphatephosphate; fluorouracil; fluorocitabine; fosquidone; fostriecin sodium; gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide; iimofosine; interleukin I1 (including recombinant interleukin II or rlL.sub.2), interferon α-2a; interferon α-2b; interferon α-n1; interferon α-n3; interferon β-1a; interferon γ-1b; iproplatin; irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolide acetate; liarozole hydrochloride hydrochloride; lometrexol sodium; lomustine; losoxantrone hydrochloride; masoprocol; maytansine; mechlorethamine hydrochloride; megestrol acetate; melengestrol acetate acetate; melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin; mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolic acidacid; nocodazoie; nogalamycin; ormaplatin; oxisuran; pegaspargase; peliomycin; pentamustine; peplomycin sulfate; perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride; plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine; procarbazine hydrochloride; puromycin; puromycin hydrochloride hydrochloride; pyrazofurin; riboprine; rogletimide; safingol; safingol hydrochloride; semustine; simtrazene; sparrosate sodium; sparsomycin; spirogermanium hydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin; sulofenur; talisomycin; tecogalan sodium; tegafur; teloxantrone hydrochloride hydrochloride; temoporfin; teniposide; teroxirone; testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin; tirapazamine; toremifene citrate; trestolone acetateacetate; triciribine phosphate; trimetrexate; trimetrexate glucuronate; triptorelin; tubulozole hydrochloride; uracil mustard; uredepa; vapreotide; verteporfin; vinblastine sulfate; vincristine sulfate; vindesine sulfate; vinepidine sulfate; vinglycinate sulfate; vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate; vinzolidine sulfate sulfate); vorozole; zeniplatin; zinostatin; daunorubicin hydrochloride, agents that arrest cells in the G2-M phase and / or modulate the formation or stability of microtubules (e.g., Taxol.TM (i.e., paclitaxel), Taxotere.TM, compounds containing a taxane backbone, erbulolazole (i.e., R-55104), dolastatin 10 (i.e., DLS-10 and NSC-376128), mivobulinisethionate (i.e., as CI-980), vincristine, NSC-639829, scutellarin (i.e., as NVP-X X-A-296), ABT-751 (Abbott, i.e., E-7010), Altorhyrtins (e.g., Atorhyrtin A and Atorhyrtin C), Spongistatins (e.g., Spongistatin 1, Spongistatin 2, Spongistatin 3, Spongistatin 4, Spongistatin 5, Spongistatin 6, Spongistatin 7, Spongistatin 8, and Spongistatin 9), Cemadotin hydrochloride (i.e., LU-103793 and NSC-D-669356), Epothilones (e.g., Epothilone A, Epothilone B, Epothilone C (i.e., deoxyepothilone A or dEpoA), Epothilone D (i.e., KOS-862, dEpoB, and deoxyepothilone B), Epothilone E, Epothilone F, Epothilone B N-oxide, Epothilone AN-oxide, 16-aza-epothilone B, 21-aminoepothilone B (i.e., BMS-310705), 21-hydroxyepothilone D (i.e., deoxyepothilone F and dEpoF), 26-fluoroepothilone), auristatin PE (i.e., NSC-654663), solidotin (i.e., TZT-1027), LS-4559-P (Pharmacia, i.e., LS-4577), LS-4578 (Phar macia, LS-477-P), LS-4477 (Pharmacia), LS-4559 (Pharmacia), RPR-112378 (Aventis), vincristine sulfate, DZ-3358 (Daiichi), FR-182877 (Fujisawa, WS-9885B), GS-164 (Takeda), GS-198 (Takeda), KAR-2 (Hungarian Academy of Sciences), BSF-223651 (BASF, i.e., ILX-651 and LU-223651), SAH-49960 (Lilly / Novartis), SDZ-268970 (Lilly / Novartis), AM-97 (Armad / Kyowa Hakko), AM-132 (Armad), AM-138 (Armad / Kyowa Hakko), IDN-5005 (Indena), Nostoc 52 (i.e., LY-355703), AC-7739 (Ajinomoto, i.e., AVE-8063A and CS-39.HCl), AC-7700 (Ajinomoto, i.e., AVE-8062, AVE-8062A, CS-39-L-Ser.HCl and RPR-258062A), Vitilevuamide, Tubulysin A ( A), Canadensol, cyanidin (i.e., NSC-106969), T-138067 (Tularik, i.e., T-67, TL-138067, and TI-138067), COBRA-1 (Parker Hughes Institute, i.e., DDE-261 and WHI-261), H10 (Kansas State University), H16 (Kansas State University), Oncocidin A1 (i.e., BTO-956 and DIME), DDE-313 (Parker Hughes Institute), Fijianolide B (B), Laulimalide, SPA-2 (Parker Hughes Institute), SPA-1 (Parker Hughes Institute, i.e., SPIKET-P), 3-IAABU (Cytoskeleton / Mt.Sinai School of Medicine, i.e., MF-569), Narcosine (also known as NSC-5366), Nascapine, D-24851 (Asta Medica), A-105972 (Abbott), Hemiasterlin, 3-BAABU (Cytoskeleton / Mt.Sinai School of Medicine, i.e., MF-191), TMPN (Arizona State University), Vanadoceneacetylacetonate, T-138026 (Tularik), Monsatrol, lnanocine (i.e., NSC-698666), 3-IAABE (Cytoskeleton / Mt. Sinai School of Medicine), A-204197 (Abbott), T-607 (Tuiarik, i.e., T-900607), RPR-115781 (Aventis), Eleutherobins (such as Desmethyleleutherobin, Desaetyleleutherobin, Isoeleutherobin A, and Z-Eleutherobin), Caribaeside, Caribaeolin, Halichondrin B, D-64131 (Asta Medica), D-68144 (Asta Medica), diazonamide A, A-293620 (Abbott), NPI-2350 (Nereus), diazonamide A, TUB-245 (Aventis), A-259754 (Abbott), diozostatin, (-)-phenylalanistin (i.e., NSCL-96F037), D-68838 (AstaMedica), D-68836 (AstaMedica), myofibrillar protein B, D-43411 (Zentaris, i.e., D-81862), A-289099 (Abbott), A-318315 (Abbott), HTI-286 (i.e., SPA-110, trifluoroacetate) (Wyeth), D-82317 (Zentaris), D-82318 (Zentaris), SC-12983 (NCI), Resverastatin phosphate sodium, BPR-OY-007 (National Health Research Institutes) and SSR-250411 (Sanofi)), steroids (e.g., dexamethasone), finasteride, aromatase inhibitors, gonadotropin-releasing hormone agonists (GnRH) such as goserelin or leuprolide, adrenocortical steroids (e.g., prednisone), progestins (e.g., hydroxyprogesterone caproate, megestrol acetate, medroxyprogesterone acetate), estrogens (e.g., diethylstilbestrol, ethinyl estradiol), antiestrogens (e.g., tamoxifen), androgens (e.g., testosterone propionate, fluoxymesterone), antiandrogens (e.g., flutamide), immunostimulants (e.g., bacillus Calmette-Guérin (BCG), levamisole, interleukin-2, alpha-interferon, etc.), monoclonal antibodies (e.g., anti-CD20, anti-HER2, anti-CD52, anti- HLA-DR and anti-VEGF monoclonal antibodies), immunotoxins (e.g., anti-CD33 monoclonal antibody-calicheamicin conjugate, anti-CD22 monoclonal antibody-Pseudomonas exotoxin conjugate, etc.), radioimmunotherapy (e.g., anti-CD20 monoclonal antibody conjugated with 111In, 90Y or 131I, etc.), triptolide, homoharringtonine, actinomycin D, doxorubicin, epirubicin, topotecan, itraconazole, vindesine, cerivastatin, vincristine, deoxyadenosine, sertraline, pitavastatin, irinotecan, clofazimine, 5-nonyloxytryptamine, vemurafenib, dabrafenib, erlotinib, gefitinib, EGFR inhibitors, epidermal growth factor receptor (EGFR) targeted therapy or therapeutic agents (e.g., gefitinib (Iressa TM ), erlotinib (Tarceva TM ), Erbitux TM ), lapatinib (Tykerb TM ), panitumumab (Vectibix TM ), Vandetanib (Caprelsa TM), afatinib / BIBW2992, CI-1033 / canertinib, neratinib / HKI-272, CP-724714, TAK-285, AST-1306, ARRY334543, ARRY-380, AG-1478, dacomitinib / PF299804, OSI-420 / desmethylerlotinib, AZD8931, AEE788, pelitinib / EKB-569, CUDC-101, WZ8040, WZ4002, WZ3146, AG-490, XL647, PD153035, BMS-599626), sorafenib, imatinib, sunitinib, dasatinib, etc.

[0140] Formulations suitable for oral administration may consist of: (a) a liquid solution, such as an effective amount of an antibody provided herein suspended in a diluent such as water, saline or PEG 400; (b) a capsule, sachet or tablet, each containing a predetermined amount of active ingredient, such as a liquid, solid, granules or gelatin; (c) a suspension in an appropriate liquid; and (d) a suitable emulsion. Tablet forms may include one or more of the following: lactose, sucrose, mannitol, sorbitol, calcium phosphate, corn starch, potato starch, microcrystalline cellulose, gelatin, colloidal silicon dioxide, talc, magnesium stearate, stearic acid, and other excipients, colorants, fillers, binders, diluents, buffers, wetting agents, preservatives, flavorings, dyes, disintegrants and pharmaceutically compatible carriers. Lozenge forms may contain the active ingredient in a flavoring, for example, sucrose, as well as lozenges containing the active ingredient in an inert base such as gelatin and glycerin or sucrose and acacia emulsions, gels, and the like which contain in addition to the active ingredient such carriers as are known in the art.

[0141] Pharmaceutical compositions may also include large, slowly metabolized macromolecules such as proteins, polysaccharides such as chitosan, polylactic acid, polyglycolic acid and copolymers (such as latex functionalized sepharose (TM), agarose, cellulose, etc.), polymeric amino acids, amino acid copolymers and lipid aggregates (such as oil droplets or liposomes). In addition, these carriers can be used as immunopotentiators (i.e., adjuvants).

[0142] Suitable formulations for rectal administration include, for example, suppositories consisting of the packaged nucleic acid with a suppository base. Suitable suppository bases include natural or synthetic triglycerides or paraffin hydrocarbons. In addition, gelatin rectal capsules may also be used, consisting of a combination of the selected compound and a base including, for example, liquid triglycerides, polyethylene glycols, and paraffin hydrocarbons.

[0143] Preparations suitable for parenteral administration, such as, for example, preparations administered by intraarticular (intra-articular), intravenous, intramuscular, intratumoral, intradermal, intraperitoneal and subcutaneous routes, include aqueous and non-aqueous isotonic sterile injection solutions, which may contain antioxidants, buffers, antibacterial agents and solutes that make the preparation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions, which may include suspending agents, solubilizers, thickeners, stabilizers and preservatives. In the practice of the present invention, the composition can be, for example, administered by intravenous infusion, oral, topical, intraperitoneal, intravesical or intrathecal form. Parenteral administration, oral administration and intravenous administration are all preferred methods of administration. The compound preparation can be provided in unit dose or multi-dose sealed containers (e.g., ampoules and vials).

[0144] Injection solutions and suspensions can be prepared from the above-described sterile powders, granules and tablets. As described above, cells transduced with nucleic acids for use in ex vivo therapy can also be administered in intravenous or parenteral form.

[0145] The pharmaceutical formulation is preferably in unit dosage form. In this form, the formulation is subdivided into unit doses containing appropriate quantities of the active ingredient. The unit dosage form can be a packaged formulation containing discrete quantities of the formulation, such as packaged tablets, capsules, and powders in vials or ampoules. Likewise, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be an appropriate number of any of these packaged forms. The composition can also contain other compatible therapeutic agents, if desired.

[0146] Co-administration encompasses combined administration using separate formulations or a single pharmaceutical formulation, as well as consecutive administration in either order, wherein preferably over a period of time, both (or all) active agents simultaneously exert their biological activities.

[0147] The effective dosage of the compositions provided herein varies depending on many different factors, including the mode of administration, the target site, the physiological state of the patient, whether the patient is human or animal, other drugs being administered, and whether the treatment is preventive or therapeutic. However, one of ordinary skill in the art will immediately recognize appropriate and / or equivalent dosages after understanding the dosages of the approved compositions for the treatment and prevention of cancer for guidance.

[0148] "Pharmaceutically acceptable excipients" and "pharmaceutically acceptable carriers" refer to substances that facilitate administration of active agents to subjects and facilitate absorption by subjects, and can be included in the compositions of the present invention without causing significant adverse toxicological effects to patients. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, physiological saline solutions, lactated Ringer's solution, ordinary sucrose, ordinary glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavoring agents, salt solutions (e.g., Ringer's solution), alcohols, oils, gelatin, carbohydrates (e.g., lactose, amylose or starch), fatty acid esters, carboxymethyl cellulose, polyvinyl pyrrolidone, and pigments, etc. Such preparations can be sterilized and, if necessary, can be mixed with adjuvants (such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for affecting osmotic pressure, buffers, coloring substances, and / or aromatic substances, etc.), and the adjuvants do not react harmfully with the compounds of the present invention. Those skilled in the art will recognize that other pharmaceutical excipients can be used in the present invention.

[0149] The term "pharmaceutically acceptable salt" refers to salts derived from a variety of organic and inorganic counterions well known in the art, and include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like; and when the molecule contains a basic functional group, salts of organic or inorganic acids such as hydrochloride, hydrobromide, tartrate, methanesulfonate, acetate, maleate, oxalate, and the like.

[0150] The term "preparation" is intended to include the formulation of the active compound with encapsulating material as a carrier, the encapsulating material providing a capsule in which the active ingredient, with or without other carriers, is surrounded by the carrier so associated therewith. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

[0151] The pharmaceutical formulation is optionally in unit dosage form. In this form, the formulation is subdivided into unit doses containing appropriate quantities of the active ingredient. The unit dosage form can be a packaged formulation containing discrete quantities of the formulation, such as packaged tablets, capsules, and powders in vials or ampoules. Likewise, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be an appropriate number of any of these packaged forms. The unit dosage form can be a frozen dispersion.

[0152] The compositions of the present invention may additionally comprise components that provide sustained release and / or comfort. Such components include high molecular weight, anionic mucoid polymers, gelling polysaccharides, and finely divided drug carrier substrates. These components are discussed in more detail in U.S. Patents 4,911,920; 5,403,841; 5,212,162; and 4,861,760. The entire contents of these patents are incorporated herein by reference in their entirety for all purposes. The compositions of the present invention can also be delivered in the form of microspheres for slow release in vivo. For example, the microspheres can be administered via intradermal injection of drug-containing microspheres that are slowly released subcutaneously (see Rao, J. Biomater Sci. Polym. Ed. 7:623-645, 1995; administered as a biodegradable and injectable gel formulation (see, e.g., Gao et al., 2001). Pharm. Res. 12: 857-863, 1995); or as microspheres for oral administration (see, e.g., Eyles, J. Pharm. Pharmacol. 49: 669-674, 1997). In an embodiment, the formulation of the composition of the present invention can be delivered by using liposomes that fuse with the cell membrane or are endocytosed, that is, by using receptor ligands bound to liposomes, which bind to surface membrane protein receptors to cause endocytosis. By using liposomes, especially carrying on the surface of the liposomes a ligand that is specific for the target cell or is endocytosed In other cases where the receptor ligands of a specific organ are preferentially targeted, the composition of the present invention can be delivered to the target cells in vivo. (See, for example, Al-Muhammed, J. Microencapsul. 13: 293-306, 1996; Chonn, Curr. Opin. Biotechnol. 6: 698-708, 1995; Ostro, Am. J. Hosp. Pharm. 46: 1576-1587, 1989). The composition of the present invention can also be delivered as nanoparticles.

[0153] It should be understood that the examples and embodiments described herein are for illustrative purposes only, and that various modifications or changes in view of them will be suggested to those skilled in the art, and will be included within the spirit and scope of this application and the scope of the appended claims. All publications, patents, and patent applications cited herein are incorporated herein by reference in their entirety for all purposes.

[0154] It should be understood that the examples and embodiments described herein are for illustrative purposes only, and that various modifications or changes in view of them will be suggested to those skilled in the art, and will be included within the spirit and scope of this application and the scope of the appended claims. All publications, patents, and patent applications cited herein are incorporated herein by reference in their entirety for all purposes.

[0155] Anti-CD84 antibody

[0156] Antibodies capable of binding to cluster of differentiation 84 (CD84) are particularly provided herein. The antibodies provided herein include novel light chain and heavy chain sequences and bind to CD84 with high efficiency and specificity. Based on these binding characteristics, the antibodies provided herein are capable of effectively delivering antibody-dependent cellular toxicity (ADCC) in the cancer microenvironment, which results in cell-mediated immune defense, whereby effector cells of the immune system actively lyse cancer cells expressing CD84. Therefore, CD84 antibodies (including embodiments thereof) as provided herein can be used for anti-cancer therapeutic purposes.

[0157] In one aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:75, CDR H2 as shown in SEQ ID NO:76, and CDR H3 as shown in SEQ ID NO:77; and wherein the light chain variable domain comprises CDRL1 as shown in SEQ ID NO:78, CDR L2 as shown in SEQ ID NO:79, and CDR L3 as shown in SEQ ID NO:80.

[0158] In another aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises: a CDR H1 comprising the sequence as shown in SEQ ID NO:75, a CDR H2 comprising the sequence as shown in SEQ ID NO:76, and a CDR H3 comprising the sequence as shown in SEQ ID NO:77; and wherein the light chain variable domain comprises: a CDR L1 comprising the sequence as shown in SEQ ID NO:78, a CDR L2 comprising the sequence as shown in SEQ ID NO:79, and a CDR L3 comprising the sequence as shown in SEQ ID NO:80.

[0159] In an embodiment, the anti-CD84 antibody is a chimeric antibody. In an embodiment, the anti-CD84 antibody is a humanized antibody.

[0160] In an embodiment, the heavy chain variable domain includes FR H1 as shown in SEQ ID NO: 81, FR H2 as shown in SEQ ID NO: 82, FR H3 as shown in SEQ ID NO: 83, and FR H4 as shown in SEQ ID NO: 84. In an embodiment, the light chain variable domain includes FR L1 as shown in SEQ ID NO: 85, FR L2 as shown in SEQ ID NO: 86, FR L3 as shown in SEQ ID NO: 87, and FR L4 as shown in SEQ ID NO: 88.

[0161] In an embodiment, the heavy chain variable domain comprises: FR H1 comprising the sequence shown in SEQ ID NO: 81, FR H2 comprising the sequence shown in SEQ ID NO: 82, FR H3 comprising the sequence shown in SEQ ID NO: 83, and FR H4 comprising the sequence shown in SEQ ID NO: 84. In an embodiment, the light chain variable domain comprises: FR L1 comprising the sequence shown in SEQ ID NO: 85, FR L2 comprising the sequence shown in SEQ ID NO: 86, FR L3 comprising the sequence shown in SEQ ID NO: 87, and FR L4 comprising the sequence shown in SEQ ID NO: 88.

[0162] In an embodiment, the heavy chain variable domain comprises the sequence of SEQ ID NO: 89. In an embodiment, the heavy chain variable domain has the sequence of SEQ ID NO: 89. In an embodiment, the light chain variable domain comprises the sequence of SEQ ID NO: 90. In an embodiment, the light chain variable domain has the sequence of SEQ ID NO: 90.

[0163] In an embodiment, the anti-CD84 antibody comprises a heavy chain comprising the sequence of SEQ ID NO: 91. In an embodiment, the anti-CD84 antibody comprises a heavy chain having the sequence of SEQ ID NO: 91. In an embodiment, the anti-CD84 antibody comprises a light chain comprising the sequence of SEQ ID NO: 92. In an embodiment, the anti-CD84 antibody comprises a light chain having the sequence of SEQ ID NO: 92. In an embodiment, the anti-CD84 antibody is referred to herein as clone 7D2.

[0164] In another aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:19, CDRH2 as shown in SEQ ID NO:20, and CDR H3 as shown in SEQ ID NO:21; and wherein the light chain variable domain comprises CDR L1 as shown in SEQ ID NO:22, CDR L2 as shown in SEQ ID NO:23, and CDR L3 as shown in SEQ ID NO:24.

[0165] In another aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises: a CDR H1 comprising the sequence as shown in SEQ ID NO: 19, a CDR H2 comprising the sequence as shown in SEQ ID NO: 20, and a CDR H3 comprising the sequence as shown in SEQ ID NO: 21; and wherein the light chain variable domain comprises: a CDR L1 comprising the sequence as shown in SEQ ID NO: 22, a CDR L comprising the sequence as shown in SEQ ID NO: 23, and a CDR L3 comprising the sequence as shown in SEQ ID NO: 24.

[0166] In an embodiment, the anti-CD84 antibody is a humanized antibody. In an embodiment, the antibody comprises a heavy chain comprising the sequence of SEQ ID NO: 37. In an embodiment, the antibody comprises a heavy chain having the sequence of SEQ ID NO: 37. In an embodiment, the antibody comprises a light chain comprising the sequence of SEQ ID NO: 38. In an embodiment, the antibody comprises a light chain having the sequence of SEQ ID NO: 38.

[0167] In an embodiment, the anti-CD84 antibody is a chimeric antibody. In an embodiment, the heavy chain variable domain includes FR H1 as shown in SEQ ID NO: 25, FR H2 as shown in SEQ ID NO: 26, FR H3 as shown in SEQ ID NO: 27, and FR H4 as shown in SEQ ID NO: 28. In an embodiment, the light chain variable domain includes FR L1 as shown in SEQ ID NO: 29, FR L2 as shown in SEQ ID NO: 30, FR L3 as shown in SEQ ID NO: 31, and FR L4 as shown in SEQ ID NO: 32. In an embodiment, the heavy chain variable domain includes: FR H1 including the sequence shown in SEQ ID NO: 25, FR H2 including the sequence shown in SEQ ID NO: 26, FR H3 including the sequence shown in SEQ ID NO: 27, and FR H4 including the sequence shown in SEQ ID NO: 28. In an embodiment, the light chain variable domain includes: FR L1 including the sequence shown as SEQ ID NO:29, FR L2 including the sequence shown as SEQ ID NO:30, FR L3 including the sequence shown as SEQ ID NO:31, and FR L4 including the sequence shown as SEQ ID NO:32.

[0168] In an embodiment, the heavy chain variable domain comprises the sequence of SEQ ID NO: 33. In an embodiment, the heavy chain variable domain has the sequence of SEQ ID NO: 33. In an embodiment, the light chain variable domain comprises the sequence of SEQ ID NO: 34. In an embodiment, the light chain variable domain has the sequence of SEQ ID NO: 34. In an embodiment, the antibody comprises a heavy chain comprising the sequence of SEQ ID NO: 35. In an embodiment, the antibody comprises a heavy chain having the sequence of SEQ ID NO: 35. In an embodiment, the antibody comprises a light chain comprising the sequence of SEQ ID NO: 36. In an embodiment, the antibody comprises a light chain having the sequence of SEQ ID NO: 36. In an embodiment, the anti-CD84 antibody is referred to herein as clone 13D11.

[0169] In another aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:39, CDRH2 as shown in SEQ ID NO:40, and CDR H3 as shown in SEQ ID NO:41; and wherein the light chain variable domain comprises CDR L1 as shown in SEQ ID NO:42, CDR L2 as shown in SEQ ID NO:43, and CDR L3 as shown in SEQ ID NO:44.

[0170] In another aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises: a CDR H1 comprising the sequence shown in SEQ ID NO:39, a CDR H2 comprising the sequence shown in SEQ ID NO:40, and a CDRH3 comprising the sequence shown in SEQ ID NO:41; and wherein the light chain variable domain comprises: a CDR L1 comprising the sequence shown in SEQ ID NO:42, a CDR L2 comprising the sequence shown in SEQ ID NO:43, and a CDR L3 comprising the sequence shown in SEQ ID NO:44.

[0171] In an embodiment, the anti-CD84 antibody is a chimeric antibody. In an embodiment, the anti-CD84 antibody is a humanized antibody. In an embodiment, the heavy chain variable domain includes FR H1 as shown in SEQ ID NO: 45, FR H2 as shown in SEQ ID NO: 46, FR H3 as shown in SEQ ID NO: 47, and FR H4 as shown in SEQ ID NO: 48. In an embodiment, the light chain variable domain includes FR L1 as shown in SEQ ID NO: 49, FR L2 as shown in SEQ ID NO: 50, FR L3 as shown in SEQ ID NO: 51, and FR L4 as shown in SEQ ID NO: 52. In an embodiment, the heavy chain variable domain includes: FR H1 including the sequence shown in SEQ ID NO: 45, FR H2 including the sequence shown in SEQ ID NO: 46, FR H3 including the sequence shown in SEQ ID NO: 47, and FR H4 including the sequence shown in SEQ ID NO: 48. In an embodiment, the light chain variable domain includes: FR L1 including the sequence shown as SEQ ID NO:49, FR L2 including the sequence shown as SEQ ID NO:50, FR L3 including the sequence shown as SEQ ID NO:51, and FR L4 including the sequence shown as SEQ ID NO:52.

[0172] In an embodiment, the heavy chain variable domain comprises the sequence of SEQ ID NO: 53. In an embodiment, the heavy chain variable domain has the sequence of SEQ ID NO: 53. In an embodiment, the light chain variable domain comprises the sequence of SEQ ID NO: 54. In an embodiment, the light chain variable domain has the sequence of SEQ ID NO: 54. In an embodiment, the anti-CD84 antibody comprises a heavy chain comprising the sequence of SEQ ID NO: 55. In an embodiment, the anti-CD84 antibody comprises a heavy chain having the sequence of SEQ ID NO: 55. In an embodiment, the anti-CD84 antibody comprises a light chain comprising the sequence of SEQ ID NO: 56. In an embodiment, the anti-CD84 antibody comprises a light chain having the sequence of SEQ ID NO: 56. In an embodiment, the anti-CD84 antibody is referred to herein as clone 2H7.

[0173] In another aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:57, CDRH2 as shown in SEQ ID NO:58, and CDR H3 as shown in SEQ ID NO:59; and wherein the light chain variable domain comprises CDR L1 as shown in SEQ ID NO:60, CDR L2 as shown in SEQ ID NO:61, and CDR L3 as shown in SEQ ID NO:62.

[0174] In another aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises: a CDR H1 comprising the sequence as shown in SEQ ID NO:57, a CDR H2 comprising the sequence as shown in SEQ ID NO:58, and a CDRH3 comprising the sequence as shown in SEQ ID NO:59; and wherein the light chain variable domain comprises: a CDR L1 comprising the sequence as shown in SEQ ID NO:60, a CDR L2 comprising the sequence as shown in SEQ ID NO:61, and a CDR L3 comprising the sequence as shown in SEQ ID NO:62.

[0175] In an embodiment, the anti-CD84 antibody is a chimeric antibody. In an embodiment, the anti-CD84 antibody is a humanized antibody. In an embodiment, the heavy chain variable domain includes FR H1 as shown in SEQ ID NO: 63, FR H2 as shown in SEQ ID NO: 64, FR H3 as shown in SEQ ID NO: 65, and FR H4 as shown in SEQ ID NO: 66. In an embodiment, the light chain variable domain includes FR L1 as shown in SEQ ID NO: 67, FR L2 as shown in SEQ ID NO: 68, FR L3 as shown in SEQ ID NO: 69, and FR L4 as shown in SEQ ID NO: 70. In an embodiment, the heavy chain variable domain includes: FR H1 including the sequence shown in SEQ ID NO: 63, FRH2 including the sequence shown in SEQ ID NO: 64, FRH3 including the sequence shown in SEQ ID NO: 65, and FR H4 including the sequence shown in SEQ ID NO: 66. In an embodiment, the light chain variable domain includes: FR L1 including the sequence shown as SEQ ID NO:67, FR L2 including the sequence shown as SEQ ID NO:68, FR L3 including the sequence shown as SEQ ID NO:69, and FR L4 including the sequence shown as SEQ ID NO:70.

[0176] In an embodiment, the heavy chain variable domain comprises the sequence of SEQ ID NO: 71. In an embodiment, the heavy chain variable domain has the sequence of SEQ ID NO: 71. In an embodiment, the light chain variable domain comprises the sequence of SEQ ID NO: 72. In an embodiment, the light chain variable domain has the sequence of SEQ ID NO: 72. In an embodiment, the anti-CD84 antibody comprises a heavy chain comprising the sequence of SEQ ID NO: 73. In an embodiment, the anti-CD84 antibody comprises a heavy chain having the sequence of SEQ ID NO: 73. In an embodiment, the anti-CD84 antibody comprises a light chain comprising the sequence of SEQ ID NO: 74. In an embodiment, the anti-CD84 antibody comprises a light chain having the sequence of SEQ ID NO: 74. In an embodiment, the anti-CD84 antibody is referred to herein as clone 5C6.

[0177] In another aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:93, CDRH2 as shown in SEQ ID NO:94, and CDR H3 as shown in SEQ ID NO:95; and wherein the light chain variable domain comprises CDR L1 as shown in SEQ ID NO:96, CDR L2 as shown in SEQ ID NO:97, and CDR L3 as shown in SEQ ID NO:98.

[0178] In another aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises: a CDR H1 comprising the sequence as shown in SEQ ID NO:93, a CDR H2 comprising the sequence as shown in SEQ ID NO:94, and a CDR H3 comprising the sequence as shown in SEQ ID NO:95; and wherein the light chain variable domain comprises: a CDR L1 comprising the sequence as shown in SEQ ID NO:96, a CDR L2 comprising the sequence as shown in SEQ ID NO:97, and a CDR L3 comprising the sequence as shown in SEQ ID NO:98.

[0179] In an embodiment, the anti-CD84 antibody is a chimeric antibody. In an embodiment, the anti-CD84 antibody is a humanized antibody. In an embodiment, the heavy chain variable domain includes FR H1 as shown in SEQ ID NO: 99, FR H2 as shown in SEQ ID NO: 100, FR H3 as shown in SEQ ID NO: 101, and FR H4 as shown in SEQ ID NO: 102. In an embodiment, the light chain variable domain includes FR L1 as shown in SEQ ID NO: 103, FR L2 as shown in SEQ ID NO: 104, FR L3 as shown in SEQ ID NO: 105, and FR L4 as shown in SEQ ID NO: 106. In an embodiment, the heavy chain variable domain includes: FR H1 including the sequence shown in SEQ ID NO: 99, FR H2 including the sequence shown in SEQ ID NO: 100, FR H3 including the sequence shown in SEQ ID NO: 101, and FRH4 including the sequence shown in SEQ ID NO: 102. In an embodiment, the light chain variable domain includes: FR L1 including the sequence shown as SEQ ID NO:103, FR L2 including the sequence shown as SEQ ID NO:104, FR L3 including the sequence shown as SEQ ID NO:105, and FR L4 including the sequence shown as SEQ ID NO:106.

[0180] In an embodiment, the heavy chain variable domain comprises the sequence of SEQ ID NO: 107. In an embodiment, the heavy chain variable domain has the sequence of SEQ ID NO: 107. In an embodiment, the light chain variable domain comprises the sequence of SEQ ID NO: 108. In an embodiment, the light chain variable domain has the sequence of SEQ ID NO: 108. In an embodiment, the anti-CD84 antibody comprises a heavy chain comprising the sequence of SEQ ID NO: 109. In an embodiment, the anti-CD84 antibody comprises a heavy chain having the sequence of SEQ ID NO: 109. In an embodiment, the anti-CD84 antibody comprises a light chain comprising the sequence of SEQ ID NO: 110. In an embodiment, the anti-CD84 antibody comprises a light chain having the sequence of SEQ ID NO: 110. In an embodiment, the anti-CD84 antibody is referred to herein as clone 8A5.

[0181] In another aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:1, CDR H2 as shown in SEQ ID NO:2, and CDR H3 as shown in SEQ ID NO:3; and wherein the light chain variable domain comprises CDRL1 as shown in SEQ ID NO:4, CDR L2 as shown in SEQ ID NO:5, and CDR L3 as shown in SEQ ID NO:6.

[0182] In another aspect, an anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain is provided, wherein the heavy chain variable domain comprises: a CDR H1 comprising the sequence shown in SEQ ID NO: 1, a CDR H2 comprising the sequence shown in SEQ ID NO: 2, and a CDR H3 comprising the sequence shown in SEQ ID NO: 3; and wherein the light chain variable domain comprises: a CDR L1 comprising the sequence shown in SEQ ID NO: 4, a CDR L2 comprising the sequence shown in SEQ ID NO: 5, and a CDR L3 comprising the sequence shown in SEQ ID NO: 6.

[0183] In an embodiment, the anti-CD84 antibody is a chimeric antibody. In an embodiment, the anti-CD84 antibody is a humanized antibody. In an embodiment, the heavy chain variable domain includes FR H1 as shown in SEQ ID NO:7, FR H2 as shown in SEQ ID NO:8, FR H3 as shown in SEQ ID NO:9, and FR H4 as shown in SEQ ID NO:10. In an embodiment, the light chain variable domain includes FRL1 as shown in SEQ ID NO:11, FR L2 as shown in SEQ ID NO:12, FR L3 as shown in SEQ ID NO:13, and FR L4 as shown in SEQ ID NO:14. In an embodiment, the heavy chain variable domain includes: FR H1 including the sequence shown in SEQ ID NO:7, FR H2 including the sequence shown in SEQ ID NO:8, FR H3 including the sequence shown in SEQ ID NO:9, and FR H4 including the sequence shown in SEQ ID NO:10. In an embodiment, the light chain variable domain includes: FR L1 including the sequence shown as SEQ ID NO:11, FR L2 including the sequence shown as SEQ ID NO:12, FR L3 including the sequence shown as SEQ ID NO:13, and FR L4 including the sequence shown as SEQ ID NO:14.

[0184] In an embodiment, the heavy chain variable domain comprises the sequence of SEQ ID NO: 15. In an embodiment, the heavy chain variable domain has the sequence of SEQ ID NO: 15. In an embodiment, the light chain variable domain comprises the sequence of SEQ ID NO: 16. In an embodiment, the light chain variable domain has the sequence of SEQ ID NO: 16. In an embodiment, the anti-CD84 antibody comprises a heavy chain comprising the sequence of SEQ ID NO: 17. In an embodiment, the anti-CD84 antibody comprises a heavy chain having the sequence of SEQ ID NO: 17. In an embodiment, the anti-CD84 antibody comprises a light chain comprising the sequence of SEQ ID NO: 18. In an embodiment, the anti-CD84 antibody comprises a light chain having the sequence of SEQ ID NO: 18. In an embodiment, the anti-CD84 antibody is referred to herein as clone 4B11.

[0185] In an embodiment, the anti-CD84 antibody is a Fab' fragment. In an embodiment, the anti-CD84 antibody is an IgG. In an embodiment, the anti-CD84 antibody is an IgG1, IgG2, IgG3, or IgG4. In an embodiment, the anti-CD84 antibody is an IgG1. In an embodiment, the anti-CD84 antibody is an IgG2. In an embodiment, the anti-CD84 antibody is an IgG3. In an embodiment, the anti-CD84 antibody is an IgG4. In an embodiment, the light chain variable domain and the heavy chain variable domain form part of a scFv.

[0186] In embodiments, anti-CD84 antibodies are capable of binding to CD84 protein. In embodiments, anti-CD84 antibodies bind to CD84 protein. In embodiments, CD84 protein is human CD84 protein. In embodiments, CD84 protein forms part of a cell. In embodiments, CD84 protein is expressed on the surface of a cell. In embodiments, the cell is a cancer cell. In embodiments, the cancer cell is a leukemia cancer cell, a myeloid cancer cell, or a lymphoma cancer cell. In embodiments, the cancer cell is a leukemia cancer cell. In embodiments, the cancer cell is a myeloid cancer cell. In embodiments, the cancer cell is a lymphoma cancer cell.

[0187] In another aspect, an anti-CD84 antibody is provided, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:19, CDR H2 as shown in SEQ ID NO:20, and CDR H3 as shown in SEQ ID NO:21, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:22, CDR L2 as shown in SEQ ID NO:23, and CDR L3 as shown in SEQ ID NO:24.

[0188] In another aspect, an anti-CD84 antibody is provided, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:39, CDR H2 as shown in SEQ ID NO:40, and CDR H3 as shown in SEQ ID NO:41, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:42, CDR L2 as shown in SEQ ID NO:43, and CDR L3 as shown in SEQ ID NO:44.

[0189] In another aspect, an anti-CD84 antibody is provided, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:57, CDR H2 as shown in SEQ ID NO:58, and CDR H3 as shown in SEQ ID NO:59, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:60, CDR L2 as shown in SEQ ID NO:61, and CDR L3 as shown in SEQ ID NO:62.

[0190] In another aspect, an anti-CD84 antibody is provided, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:75, CDR H2 as shown in SEQ ID NO:76, and CDR H3 as shown in SEQ ID NO:77, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:78, CDR L2 as shown in SEQ ID NO:79, and CDR L3 as shown in SEQ ID NO:80.

[0191] In another aspect, an anti-CD84 antibody is provided, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:93, CDR H2 as shown in SEQ ID NO:94, and CDR H3 as shown in SEQ ID NO:95, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:96, CDR L2 as shown in SEQ ID NO:97, and CDR L3 as shown in SEQ ID NO:98.

[0192] In another aspect, an anti-CD84 antibody is provided, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:1, CDR H2 as shown in SEQ ID NO:2, and CDR H3 as shown in SEQ ID NO:3, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:4, CDR L2 as shown in SEQ ID NO:5, and CDR L3 as shown in SEQ ID NO:6.

[0193] In embodiments, the anti-CD84 antibody is linked to a therapeutic or diagnostic moiety. In embodiments, the anti-CD84 antibody is linked to a therapeutic moiety. In embodiments, the anti-CD84 antibody is linked to a diagnostic moiety. In embodiments, the therapeutic moiety is an anti-cancer moiety.

[0194] As described above, "light chain variable (VL) domains" as provided herein form a part of the variable region of the light chain of an antibody, antibody variant, or its fragment. Similarly, "heavy chain variable (VH) domains" as provided herein form a part of the variable region of the heavy chain of an antibody, antibody variant, or its fragment. The light chain variable domains and the heavy chain variable domains together form a paratope for binding antigens (epitopes). Paratopes or antigen binding sites are formed at the N-terminus of antibodies, antibody variants, or their fragments. In embodiments, light chain variable (VL) domains include CDR L1, CDR L2, CDR L3, and FR L1, FR L2, FR L3, and FR L4 (framework regions) of antibody light chains. In embodiments, heavy chain variable (VH) domains include CDR H1, CDR H2, CDR H3, and FR H1, FR H2, FRH3, and FR H4 (framework regions) of antibody heavy chains. In embodiments, light chain variable (VL) domains and light chain constant (CL) domains form a part of antibody light chains. In an embodiment, a heavy chain variable (VH) domain and a heavy chain constant (CH1) domain form a part of an antibody heavy chain. In an embodiment, a heavy chain variable (VH) domain and one or more heavy chain constant (CH1, CH2 or CH3) domains form a part of an antibody heavy chain. Therefore, in an embodiment, a light chain variable (VL) domain forms a part of an antibody. In an embodiment, a heavy chain variable (VH) domain forms a part of an antibody. In an embodiment, a light chain variable (VL) domain forms a part of a therapeutic antibody. In an embodiment, a heavy chain variable (VH) domain forms a part of a therapeutic antibody. In an embodiment, a light chain variable (VL) domain forms a part of a humanized antibody. In an embodiment, a heavy chain variable (VH) domain forms a part of a humanized antibody. In an embodiment, a light chain variable (VL) domain forms a part of a humanized antibody. In an embodiment, a heavy chain variable (VH) domain forms a part of a humanized antibody. In an embodiment, a light chain variable (VL) domain forms a part of a chimeric antibody. In an embodiment, a heavy chain variable (VH) domain forms a part of a chimeric antibody. In an embodiment, the light chain variable (VL) domain forms a part of an antibody fragment. In an embodiment, the heavy chain variable (VH) domain forms a part of an antibody fragment. In an embodiment, the light chain variable (VL) domain forms a part of an antibody variant. In an embodiment, the heavy chain variable (VH) domain forms a part of an antibody variant. In an embodiment, the light chain variable (VL) domain forms a part of a Fab. In an embodiment, the heavy chain variable (VH) domain forms a part of a Fab. In an embodiment, the light chain variable (VL) domain forms a part of a scFv. In an embodiment, the heavy chain variable (VH) domain forms a part of a scFv. In an embodiment, the light chain variable (VL) domain forms a part of a single domain antibody. In an embodiment, the heavy chain variable (VH) domain forms a part of a single domain antibody.

[0195] In one embodiment, the heavy chain variable domain has the sequence of SEQ ID NO: 15 and the light chain variable domain has the sequence of SEQ ID NO: 16. In one embodiment, the anti-CD84 antibody has a heavy chain of SEQ ID NO: 17 and a light chain of SEQ ID NO: 18. In another embodiment, the antibody is 4B11.

[0196] In one embodiment, the heavy chain variable domain has the sequence of SEQ ID NO: 33 and the light chain variable domain has the sequence of SEQ ID NO: 34. In one embodiment, the anti-CD84 antibody has a heavy chain of SEQ ID NO: 35 and a light chain of SEQ ID NO: 36. In another embodiment, the antibody is 13D11.

[0197] In one embodiment, the anti-CD84 antibody has a heavy chain of SEQ ID NO: 37 and a light chain of SEQ ID NO: 38. In another embodiment, the antibody is 13D11 Hu.

[0198] In one embodiment, the heavy chain variable domain has the sequence of SEQ ID NO: 53 and the light chain variable domain has the sequence of SEQ ID NO: 54. In one embodiment, the anti-CD84 antibody has a heavy chain of SEQ ID NO: 55 and a light chain of SEQ ID NO: 56. In another embodiment, the antibody is 2H7.

[0199] In one embodiment, the heavy chain variable domain has the sequence of SEQ ID NO: 71 and the light chain variable domain has the sequence of SEQ ID NO: 72. In one embodiment, the anti-CD84 antibody has a heavy chain of SEQ ID NO: 73 and a light chain of SEQ ID NO: 74. In another embodiment, the antibody is 5C6.

[0200] In one embodiment, the heavy chain variable domain has the sequence of SEQ ID NO: 89 and the light chain variable domain has the sequence of SEQ ID NO: 90. In one embodiment, the anti-CD84 antibody has a heavy chain of SEQ ID NO: 91 and a light chain of SEQ ID NO: 92. In another embodiment, the antibody is 7D2.

[0201] In one embodiment, the heavy chain variable domain has the sequence of SEQ ID NO: 107 and the light chain variable domain has the sequence of SEQ ID NO: 108. In one embodiment, the anti-CD84 antibody has a heavy chain of SEQ ID NO: 109 and a light chain of SEQ ID NO: 110. In another embodiment, the antibody is 8A5.

[0202] CD84 epitope

[0203] The antibodies provided herein (including embodiments thereof) are capable of specifically binding to a CD84 epitope. In embodiments, the epitope is a human CD84 protein.

[0204] In embodiments, the epitope interacts with the heavy chain of an anti-CD84 antibody. In an embodiment, the epitope includes Val at a position corresponding to position 21 of SEQ ID NO: 111, Asn at a position corresponding to position 22 of SEQ ID NO: 111, Ile at a position corresponding to position 23 of SEQ ID NO: 111, Gln at a position corresponding to position 24 of SEQ ID NO: 111, Glu at a position corresponding to position 25 of SEQ ID NO: 111, Pro at a position corresponding to position 26 of SEQ ID NO: 111, Arg at a position corresponding to position 27 of SEQ ID NO: 111, Gln at a position corresponding to position 28 of SEQ ID NO: 111, Ser at a position corresponding to position 48 of SEQ ID NO: 111, Glu at a position corresponding to position 49 of SEQ ID NO: 111, Pro at a position corresponding to position 70 of SEQ ID NO: 111, Tyr at a position corresponding to position 72 of SEQ ID NO: 111, Ala at a position corresponding to position 95 of SEQ ID NO: 111, Asp at a position corresponding to position 96 of SEQ ID NO: 111, Arg at a position corresponding to position 27 of SEQ ID NO: 111, Gln at a position corresponding to position 28 of SEQ ID NO: 111, Ser at a position corresponding to position 48 of SEQ ID NO: 111, Glu at a position corresponding to position 49 of SEQ ID NO: 111, Pro at position 97 of SEQ ID NO:111, Tyr at a position corresponding to position 98 of SEQ ID NO:111, Thr at a position corresponding to position 100 of SEQ ID NO:111, or Lys at a position corresponding to position 102 of SEQ ID NO:111.

[0205] In embodiments, the epitope includes Val at a position corresponding to position 21 of SEQ ID NO: 111. In embodiments, the epitope includes Asn at a position corresponding to position 22 of SEQ ID NO: 111. In embodiments, the epitope includes Ile at a position corresponding to position 23 of SEQ ID NO: 111. In embodiments, the epitope includes Gln at a position corresponding to position 24 of SEQ ID NO: 111. In embodiments, the epitope includes Glu at a position corresponding to position 25 of SEQ ID NO: 111. In embodiments, the epitope includes Pro at a position corresponding to position 26 of SEQ ID NO: 111. In embodiments, the epitope includes Arg at a position corresponding to position 27 of SEQ ID NO: 111. In embodiments, the epitope includes Gln at a position corresponding to position 28 of SEQ ID NO: 111. In embodiments, the epitope includes Ser at a position corresponding to position 48 of SEQ ID NO: 111. In embodiments, the epitope includes Glu at a position corresponding to position 49 of SEQ ID NO: 111. In embodiments, the epitope includes Pro at a position corresponding to position 70 of SEQ ID NO: 111. In embodiments, the epitope includes Tyr at a position corresponding to position 72 of SEQ ID NO: 111. In embodiments, the epitope includes Ala at a position corresponding to position 95 of SEQ ID NO: 111. In embodiments, the epitope includes Asp at a position corresponding to position 96 of SEQ ID NO: 111. In embodiments, the epitope includes Pro at a position corresponding to position 97 of SEQ ID NO: 111. In embodiments, the epitope includes Tyr at a position corresponding to position 98 of SEQ ID NO: 111. In embodiments, the epitope includes Thr at a position corresponding to position 100 of SEQ ID NO: 111. In embodiments, the epitope includes Lys at a position corresponding to position 102 of SEQ ID NO: 111.

[0206] In an embodiment, the epitope includes Val at a position corresponding to position 21 of SEQ ID NO: 111, Asn at a position corresponding to position 22 of SEQ ID NO: 111, Ile at a position corresponding to position 23 of SEQ ID NO: 111, Gln at a position corresponding to position 24 of SEQ ID NO: 111, Glu at a position corresponding to position 25 of SEQ ID NO: 111, Pro at a position corresponding to position 26 of SEQ ID NO: 111, Arg at a position corresponding to position 27 of SEQ ID NO: 111, Gln at a position corresponding to position 28 of SEQ ID NO: 111, Ser at a position corresponding to position 48 of SEQ ID NO: 111, Glu at a position corresponding to position 49 of SEQ ID NO: 111, Pro at a position corresponding to position 70 of SEQ ID NO: 111, Tyr at a position corresponding to position 72 of SEQ ID NO: 111, Ala at a position corresponding to position 95 of SEQ ID NO: 111, Asp at a position corresponding to position 96 of SEQ ID NO: 111, Arg at a position corresponding to position 27 of SEQ ID NO: 111, Gln at a position corresponding to position 28 of SEQ ID NO: 111, Ser at a position corresponding to position 48 of SEQ ID NO: 111, Glu at a position corresponding to position 49 of SEQ ID NO: 111, Pro at position 97 of SEQ ID NO:111, Tyr at a position corresponding to position 98 of SEQ ID NO:111, Thr at a position corresponding to position 100 of SEQ ID NO:111, and Lys at a position corresponding to position 102 of SEQ ID NO:111.

[0207] In one embodiment, the heavy chain of the anti-CD84 antibody binds to an epitope having Thr at a position corresponding to position 30 of SEQ ID NO: 109, Thr at a position corresponding to position 31 of SEQ ID NO: 109, Tyr at a position corresponding to position 32 of SEQ ID NO: 109, Ser at a position corresponding to position 33 of SEQ ID NO: 109, His at a position corresponding to position 35 of SEQ ID NO: 109, Trp at a position corresponding to position 47 of SEQ ID NO: 109, Trp at a position corresponding to position 50 of SEQ ID NO: 109, Asp at a position corresponding to position 52 of SEQ ID NO: 109, Thr at a position corresponding to position 53 of SEQ ID NO: 109, Ala at a position corresponding to position 54 of SEQ ID NO: 109, Thr at a position corresponding to position 55 of SEQ ID NO: 109, Glu at a position corresponding to position 57 of SEQ ID NO: 109, Pro at a position corresponding to position 58 of SEQ ID NO: 109, Thr at the position corresponding to position 59 of SEQ ID NO: 109, Tyr at the position corresponding to position 60 of SEQ ID NO: 109, Ala at the position corresponding to position 61 of SEQ ID NO: 109, Lys at the position corresponding to position 65 of SEQ ID NO: 109, Ser at the position corresponding to position 99 of SEQ ID NO: 109, Tyr at the position corresponding to position 101 of SEQ ID NO: 109, Trp at the position corresponding to position 102 of SEQ ID NO: 109, Tyr at the position corresponding to position 103 of SEQ ID NO: 109, or Phe at the position corresponding to position 104 of SEQ ID NO: 109.

[0208] In another embodiment, the heavy chain of the anti-CD84 antibody binds to an epitope having Thr at a position corresponding to position 30 of SEQ ID NO: 109, Thr at a position corresponding to position 31 of SEQ ID NO: 109, Tyr at a position corresponding to position 32 of SEQ ID NO: 109, Ser at a position corresponding to position 33 of SEQ ID NO: 109, His at a position corresponding to position 35 of SEQ ID NO: 109, Trp at a position corresponding to position 47 of SEQ ID NO: 109, Trp at a position corresponding to position 50 of SEQ ID NO: 109, Asp at a position corresponding to position 52 of SEQ ID NO: 109, Thr at a position corresponding to position 53 of SEQ ID NO: 109, Ala at a position corresponding to position 54 of SEQ ID NO: 109, Thr at a position corresponding to position 55 of SEQ ID NO: 109, Glu at a position corresponding to position 57 of SEQ ID NO: 109, Pro at a position corresponding to position 58 of SEQ ID NO: 109, Thr at the position corresponding to position 59 of SEQ ID NO: 109, Tyr at the position corresponding to position 60 of SEQ ID NO: 109, Ala at the position corresponding to position 61 of SEQ ID NO: 109, Lys at the position corresponding to position 65 of SEQ ID NO: 109, Ser at the position corresponding to position 99 of SEQ ID NO: 109, Tyr at the position corresponding to position 101 of SEQ ID NO: 109, Trp at the position corresponding to position 102 of SEQ ID NO: 109, Tyr at the position corresponding to position 103 of SEQ ID NO: 109, and Phe at the position corresponding to position 104 of SEQ ID NO: 109.

[0209] In one embodiment, the heavy chain of the anti-CD84 antibody binds to Gln at a position corresponding to position 24 of SEQ ID NO: 111 and to Ser at a position corresponding to position 99 of SEQ ID NO: 109. In one embodiment, the heavy chain of the anti-CD84 antibody binds to Gln at a position corresponding to position 24 of SEQ ID NO: 111 and to Thr at a position corresponding to position 53 of SEQ ID NO: 109. In one embodiment, the heavy chain of the anti-CD84 antibody binds to Glu at a position corresponding to position 25 of SEQ ID NO: 111 and to Ser at a position corresponding to position 33 of SEQ ID NO: 109. In one embodiment, the heavy chain of the anti-CD84 antibody binds to Gln at a position corresponding to position 24 of SEQ ID NO: 111 and to Thr at a position corresponding to position 30 of SEQ ID NO: 109.

[0210] In embodiments, the epitope interacts with the light chain of an anti-CD84 antibody. In embodiments, the epitope includes Glu at a position corresponding to position 25 of SEQ ID NO: 111, Arg at a position corresponding to position 27 of SEQ ID NO: 111, Gln at a position corresponding to position 28 of SEQ ID NO: 111, Val at a position corresponding to position 29 of SEQ ID NO: 111, Lys at a position corresponding to position 30 of SEQ ID NO: 111, Pro at a position corresponding to position 45 of SEQ ID NO: 111, Gly at a position corresponding to position 46 of SEQ ID NO: 111, Ser at a position corresponding to position 48 of SEQ ID NO: 111, Glu at a position corresponding to position 49 of SEQ ID NO: 111, Gln at a position corresponding to position 94 of SEQ ID NO: 111, or Ala at a position corresponding to position 95 of SEQ ID NO: 111.

[0211] In embodiments, the epitope includes Glu at a position corresponding to position 25 of SEQ ID NO: 111. In embodiments, the epitope includes Arg at a position corresponding to position 27 of SEQ ID NO: 111. In embodiments, the epitope includes Gln at a position corresponding to position 28 of SEQ ID NO: 111. In embodiments, the epitope includes Val at a position corresponding to position 29 of SEQ ID NO: 111. In embodiments, the epitope includes Lys at a position corresponding to position 30 of SEQ ID NO: 111. In embodiments, the epitope includes Pro at a position corresponding to position 45 of SEQ ID NO: 111. In embodiments, the epitope includes Gly at a position corresponding to position 46 of SEQ ID NO: 111. In embodiments, the epitope includes Ser at a position corresponding to position 48 of SEQ ID NO: 111. In embodiments, the epitope includes Glu at a position corresponding to position 49 of SEQ ID NO: 111. In embodiments, the epitope includes Gln at a position corresponding to position 94 of SEQ ID NO: 111. In embodiments, the epitope includes Ala at a position corresponding to position 95 of SEQ ID NO: 111.

[0212] In an embodiment, the epitope includes Glu at a position corresponding to position 25 of SEQ ID NO: 111, Arg at a position corresponding to position 27 of SEQ ID NO: 111, Gln at a position corresponding to position 28 of SEQ ID NO: 111, Val at a position corresponding to position 29 of SEQ ID NO: 111, Lys at a position corresponding to position 30 of SEQ ID NO: 111, Pro at a position corresponding to position 45 of SEQ ID NO: 111, Gly at a position corresponding to position 46 of SEQ ID NO: 111, Ser at a position corresponding to position 48 of SEQ ID NO: 111, Glu at a position corresponding to position 49 of SEQ ID NO: 111, Gln at a position corresponding to position 94 of SEQ ID NO: 111, and Ala at a position corresponding to position 95 of SEQ ID NO: 111.

[0213] In one embodiment, the light chain of an anti-CD84 antibody binds to an epitope having His at a position corresponding to position 31 of SEQ ID NO: 110, Ile at a position corresponding to position 32 of SEQ ID NO: 110, Asn at a position corresponding to position 33 of SEQ ID NO: 110, Gly at a position corresponding to position 34 of SEQ ID NO: 110, Asn at a position corresponding to position 35 of SEQ ID NO: 110, Tyr at a position corresponding to position 37 of SEQ ID NO: 110, Glu at a position corresponding to position 39 of SEQ ID NO: 110, Lys at a position corresponding to position 55 of SEQ ID NO: 110, Phe at a position corresponding to position 94 of SEQ ID NO: 110, Gly at a position corresponding to position 96 of SEQ ID NO: 110, Ser at a position corresponding to position 97 of SEQ ID NO: 110, His at a position corresponding to position 98 of SEQ ID NO: 110, Val at a position corresponding to position 99 of SEQ ID NO: 110, or a position corresponding to position 91 of SEQ ID NO: 110. Trp at position 101 of ID NO:110.

[0214] In one embodiment, the light chain of an anti-CD84 antibody binds to an epitope having His at a position corresponding to position 31 of SEQ ID NO: 110, Ile at a position corresponding to position 32 of SEQ ID NO: 110, Asn at a position corresponding to position 33 of SEQ ID NO: 110, Gly at a position corresponding to position 34 of SEQ ID NO: 110, Asn at a position corresponding to position 35 of SEQ ID NO: 110, Tyr at a position corresponding to position 37 of SEQ ID NO: 110, Glu at a position corresponding to position 39 of SEQ ID NO: 110, Lys at a position corresponding to position 55 of SEQ ID NO: 110, Phe at a position corresponding to position 94 of SEQ ID NO: 110, Gly at a position corresponding to position 96 of SEQ ID NO: 110, Ser at a position corresponding to position 97 of SEQ ID NO: 110, His at a position corresponding to position 98 of SEQ ID NO: 110, Val at a position corresponding to position 99 of SEQ ID NO: 110, and Trp at position 101 of ID NO:110.

[0215] In one embodiment, the light chain of the anti-CD84 antibody binds to Ser at a position corresponding to position 48 of SEQ ID NO: 111 and to Asn at a position corresponding to position 33 of SEQ ID NO: 110. In one embodiment, the light chain of the anti-CD84 antibody binds to Glu at a position corresponding to position 49 of SEQ ID NO: 111 and to Lys at a position corresponding to position 55 of SEQ ID NO: 110. In one embodiment, the light chain of the anti-CD84 antibody binds to Arg at a position corresponding to position 27 of SEQ ID NO: 111 and to Gly at a position corresponding to position 96 of SEQ ID NO: 110. In one embodiment, the light chain of the anti-CD84 antibody binds to Ser at a position corresponding to position 48 of SEQ ID NO: 111 and to Asn at a position corresponding to position 35 of SEQ ID NO: 110. In one embodiment, the light chain of the anti-CD84 antibody binds to Glu at a position corresponding to position 49 of SEQ ID NO: 111 and to Lys at a position corresponding to position 55 of SEQ ID NO: 110. In one embodiment, the light chain of the anti-CD84 antibody binds Arg at a position corresponding to position 27 of SEQ ID NO:111 and Glu at a position corresponding to position 39 of SEQ ID NO:110.

[0216] Nucleic acid composition

[0217] The compositions provided herein comprise nucleic acid molecules encoding anti-CD84 antibodies or portions thereof as provided herein (including embodiments thereof). Antibodies encoded by the isolated nucleic acids provided herein are described in detail throughout this application (including the descriptions above and in the Examples section). Therefore, in one aspect, isolated nucleic acids encoding anti-CD84 antibodies as provided herein (including embodiments thereof) are provided.

[0218] Cell composition

[0219] In another aspect, a cell is provided, the cell comprising an anti-CD84 antibody as provided herein (including embodiments thereof), or a nucleic acid as provided herein (including embodiments thereof).

[0220] Pharmaceutical composition

[0221] Compositions provided herein include pharmaceutical compositions comprising an anti-CD84 antibody or portion thereof as provided herein (including embodiments thereof). Thus, in one aspect, a pharmaceutical composition is provided comprising a therapeutically effective amount of an antibody provided herein (including embodiments thereof) and a pharmaceutically acceptable excipient.

[0222] Methods for producing antibodies

[0223] In one aspect, a method of generating an antibody capable of binding to CD84 is provided, the method comprising immunizing a mammal with a peptide comprising the sequence of SEQ ID NO: 111.

[0224] Treatment

[0225] The compositions provided herein (e.g., anti-CD84 antibodies), including embodiments thereof, are believed to provide effective treatment for diseases such as cancer (e.g., acute myeloid leukemia). Therefore, in one aspect, a method for treating a cancer suffered by a subject in need thereof is provided, the method comprising administering to the subject a therapeutically effective amount of an anti-CD84 antibody as provided herein (including embodiments thereof) or a pharmaceutical composition as provided herein (including embodiments thereof), thereby treating the cancer suffered by the subject. In embodiments, the cancer is a lymphoma, a myeloma, and / or a leukemia. In embodiments, the cancer is a lymphoma, a myeloma, and a leukemia. In embodiments, the cancer is a lymphoma. In embodiments, the cancer is a myeloma. In embodiments, the cancer is a leukemia. In embodiments, the cancer is acute myeloid leukemia (AML).

[0226] In embodiments, the anti-CD84 antibody is administered in an amount less than the amount of anti-cancer antibody typically administered to treat cancer. In embodiments, the amount is 1 / 2, 1 / 5, 1 / 10, 1 / 100, 1 / 200, 1 / 500, 1 / 1000, or 1 / 10000 of the amount of the anti-cancer antibody. In embodiments, the amount is 1 / 2 of the amount of the anti-cancer antibody. In embodiments, the amount is 1 / 5 of the amount of the anti-cancer antibody. In embodiments, the amount is 1 / 10 of the amount of the anti-cancer antibody. In embodiments, the amount is 1 / 100 of the amount of the anti-cancer antibody. In embodiments, the amount is 1 / 200 of the amount of the anti-cancer antibody. In embodiments, the amount is 1 / 500 of the amount of the anti-cancer antibody. In embodiments, the amount is 1 / 1000 of the amount of the anti-cancer antibody. In embodiments, the amount is 1 / 1000 of the amount of the anti-cancer antibody. In embodiments, the amount is 1 / 10000 of the amount of the anti-cancer antibody.

[0227] In an embodiment, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 1 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 1.5 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 2 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 2.5 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 3 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 3.5 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 4 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 4.5 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 5 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 5.5 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 6 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 6.5 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 7 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 7.5 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 8 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 8.5 mg / kg to about 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 9 mg / kg to about 20 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 9.5 mg / kg to about 20 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 10 mg / kg to about 20 mg / kg.

[0228] In embodiments, anti-CD84 antibodies are administered in an amount of about 10.5 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 11 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 11.5 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 12 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 12.5 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 13 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 13.5 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 14 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 14.5 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 15 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 15.5 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 6 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 16.5 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 17 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 17.5 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 18 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 18.5 mg / kg to about 20 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 19 mg / kg to about 20 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 19.5 mg / kg to about 20 mg / kg.

[0229] In embodiments, the anti-CD84 antibody is administered at about 0.5 mg / kg, 1 mg / kg, 1.5 mg / kg, 2 mg / kg, 2.5 mg / kg, 3 mg / kg, 3.5 mg / kg, 4 mg / kg, 4.5 mg / kg, 5 mg / kg, 5.5 mg / kg, 6 mg / kg, 6.5 mg / kg, 7 mg / kg, 7.5 mg / kg, 8 mg / kg, 8.5 mg / kg, 9 mg / kg, 9.5 mg / kg, 10 mg / kg, 10. In some embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg. In some embodiments, the anti-CD84 antibody is administered in an amount of about 1 mg / kg. In some embodiments, the anti-CD84 antibody is administered in an amount of about 1.5 mg / kg. In some embodiments, the anti-CD84 antibody is administered in an amount of about 2 mg / kg. In some embodiments, the anti-CD84 antibody is administered in an amount of about 2.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 3 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 3.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 4 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 4.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 5.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 6 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 6.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 7 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 7.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 8 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 8.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 9 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 9.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 10 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 10.5 mg / kg.In embodiments, the anti-CD84 antibody is administered in an amount of about 11 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 11.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 12 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 12.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 13 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 13.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 14 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 14.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 15 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 15.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 16 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 16.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 17 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 17.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 18 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 18.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 19 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 19.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 20 mg / kg.

[0230] In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 1 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 1.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 2 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 2.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 3 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 3.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 4 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 4.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 5.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 6 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 6.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 7 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 7.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 8 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 8.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 9 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 9.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 10 mg / kg to 20 mg / kg.

[0231] In an embodiment, the anti-CD84 antibody is administered in an amount of 10.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 11 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 11.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 12 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 12.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 13 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 13.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 14 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 14.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 15 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 15.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 6 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 16.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 17 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 17.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 18 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 18.5 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 19 mg / kg to 20 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 19.5 mg / kg to 20 mg / kg.

[0232] In embodiments, the anti-CD84 antibody is administered at 0.5 mg / kg, 1 mg / kg, 1.5 mg / kg, 2 mg / kg, 2.5 mg / kg, 3 mg / kg, 3.5 mg / kg, 4 mg / kg, 4.5 mg / kg, 5 mg / kg, 5.5 mg / kg, 6 mg / kg, 6.5 mg / kg, 7 mg / kg, 7.5 mg / kg, 8 mg / kg, 8.5 mg / kg, 9 mg / kg, 9.5 mg / kg, 10 mg / kg, 10. In some embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg. In some embodiments, the anti-CD84 antibody is administered in an amount of about 1 mg / kg. In some embodiments, the anti-CD84 antibody is administered in an amount of about 1.5 mg / kg. In some embodiments, the anti-CD84 antibody is administered in an amount of about 2 mg / kg. In some embodiments, the anti-CD84 antibody is administered in an amount of about 2.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 3 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 3.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 4 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 4.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 5.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 6 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 6.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 7 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 7.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 8 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 8.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 9 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 9.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 10 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 10.5 mg / kg.In embodiments, the anti-CD84 antibody is administered in an amount of about 11 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 11.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 12 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 12.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 13 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 13.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 14 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 14.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 15 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 15.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 16 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 16.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 17 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 17.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 18 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 18.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 19 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 19.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of about 20 mg / kg. As provided herein, the term "mg / kg" refers to mg of anti-CD84 antibody or a salt thereof / kg body weight.

[0233] In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 19.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 19 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 18.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 18 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 17.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 17 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 16.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 16 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 15.5 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 15 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 14.5 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 14 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 13.5 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 13 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 12.5 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 12 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 11.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 11 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 10.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 10 mg / kg.

[0234] In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 9.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 9 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 8.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 8 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 7.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 7 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 6.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 6 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of about 0.5 mg / kg to about 5.5 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 5 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 4.5 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 4 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 3.5 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 3 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 2.5 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 2 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 1.5 mg / kg. In embodiments, anti-CD84 antibodies are administered in an amount of about 0.5 mg / kg to about 1 mg / kg.

[0235] In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 19.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 19 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 18.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 18 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 17.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 17 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 16.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 16 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 15.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 15 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 14.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 14 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 13.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 13 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 12.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 12 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 11.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 11 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 10.5 mg / kg. In embodiments, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 10 mg / kg.

[0236] In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 9.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 9 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 8.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 8 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 7.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 7 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 6.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 6 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 5.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 4.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 4 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 3.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 3 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 2.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 2 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 1.5 mg / kg. In an embodiment, the anti-CD84 antibody is administered in an amount of 0.5 mg / kg to 1 mg / kg.

[0237] Example

[0238] Example 1: Production of recombinant human CD84 protein

[0239] To construct recombinant CD84 protein, the extracellular domain (ECD) of human CD84 was fused with mouse IgG2a.Fc or human IgA1.Fc ( Figure 1 The recombinant DNA encoding these two proteins was synthesized by the manufacturer (Twist Bioscience) and cloned into mammalian expression vectors. TMII plasmid Maxiprep kit (Zymo Research). CD84 fusion protein was produced using the ExpiCHO expression system (Thermo Fisher Scientific). The protocol was performed according to the manufacturer's manual. Briefly, 24 hours before DNA transfection, CHO cells were cultured at 3-4×10 6 Cells were inoculated into fresh ExpiCHO expression medium. On the day of transfection, the cells were adjusted to 6×10 6 For a 100 mL transfection, dilute 80 mg of DNA in 4 mL of OptiPRO TM SFM and then diluted in 3.7 mL OptiPRO TM 320mL ExpiFectamine in SFM TM Mix. Slowly add the mixture to the cell culture under gentle vortexing. Then incubate the cells at 37°C. After 16 to 22 hours, add 6 ml of ExpiCHO TM Enhancer and 24mL ExpiCHO TM Feed was added to the cell culture and the cells were maintained in culture at 32°C for 10 days. The culture supernatant was harvested by centrifugation at 4000×g for 30 minutes and passed through a 0.22 mm filter for protein purification. Protein A resin (GE Healthcare) and CaptureSelect TM IgA affinity matrix (Thermo Fisher Scientific) was used to purify CD84-IgG2a and CD84-IgA1, respectively. The purification procedures were performed according to the manufacturer's manual. TGX Stain-Free TM The purified proteins were run on reducing SDS-PAGE precast gels (Bio-Rad) and analyzed using a Superdex 200 Increase 10 / 300 GL column (GE Healthcare) with PBS ( Figure 2 ).

[0240] Example 2: Preparation of anti-CD84 hybridoma

[0241] All animal experiments were conducted with the approval of the Experimental Animal Care and Use Committee of the City of Hope (IUCAC #19070 and #19023). For mouse immunization, recombinant CD84-IgG2a protein was emulsified with complete Freund's adjuvant (Sigma Aldrich) and injected subcutaneously into 10 Balb / c mice (The Jackson Laboratory). Each mouse was injected with 50 micrograms of protein. Three weeks later, mice received two subcutaneous injections of 50 mg CD84-IgG2a emulsified with incomplete Freund's adjuvant (Sigma Aldrich) at intervals of two weeks. Three days before harvesting the spleen, 10 mg CD84-IgG2a was injected into mice via the tail vein. Splenocytes were harvested using PEG 1500 (Roche) for fusion with mouse myeloma cell line FO (ATCC). Cell fusion procedures were performed according to the manufacturer's manual. After fusion, the cells were cultured in a medium containing hypoxanthine / aminopterin / thymidine (Thermo Fisher Scientific) and 10% Cells were selected in complete DMEM medium with hybridoma clone supplement (Santa Cruz Biotechnology Co., Ltd.) for 10 to 12 days. The hybridoma culture supernatant was screened for the reaction of CD84-IgA1 protein using ELISA. In order to screen with ELISA, 50 mL CD84-IgA1 (pH 9.6) diluted in carbonate / bicarbonate buffer at a concentration of 1 mg / mL was added to the microwells and incubated overnight at 4°C. The wells were washed three times with PBS (PBST) containing 0.1% Tween 20 and blocked with 200 mL PBS containing 1% bovine serum albumin. After incubation at room temperature for 1 hour and washing three times with PBST, 50 mL culture supernatant was added to the wells and incubated at room temperature for 1 hour. After washing, 50 mL 1:10,000 diluted goat anti-mouse IgG.Fc-HRP (Jackson ImmunoResearch Co., Ltd.) was added to the wells and incubated at room temperature for 1 hour. After washing six times, 50 mL of TMB substrate (Thermo Fisher Scientific) was added to the wells for color development. The reaction was stopped by adding 50 mL of 1N HCl. The wells were read at an optical density of 450 nm using a Synergy 4 microplate reader (BioTek). In the ELISA screening results, 71 hybridoma clones showed strong binding signals with CD84-IgA1 protein.

[0242] Example 3: Flow cytometry of anti-CD84 hybridoma clones

[0243] Hybridoma clones selected from the ELISA screening were subjected to flow cytometric analysis using NS0 (a mouse myeloma cell line expressing human CD84 via lentiviral transduction (ECACC)). Half a million transduced cells were incubated with 100 mL of hybridoma culture supernatant on ice for 30 minutes and washed with cold PBS containing 1% fetal bovine serum and 0.1% sodium azide. The cells were then incubated with 100 mL of Alexa Fluor 5000 diluted 1:400. 488AffiniPure goat anti-mouse IgG (Jackson ImmunoResearch) was incubated on ice for 30 minutes and washed for analysis using an Accuri C6 flow cytometer (BD). The results showed that 16 clones could bind to human CD84 ( Figure 3 ). The six clones 2H7, 4B11, 5C6, 7D2, 8A5 and 13D11 (Table 1) with strong staining signals were further tested for binding to the human myeloma cell line HL-60, the T cell line Jurkat, the lymphoma cell line K-562 and the breast cell line SKBR3 (ATCC). A commercially available mouse anti-human CD84 monoclonal antibody (aCD84, BioLegend) was used as a positive control. The results showed that, consistent with the positive control, these anti-CD84 antibodies specifically bound to cells expressing CD84 and did not react with CD84-negative cells ( Figure 4 ).

[0244] Table 1 :

[0245] clone Geometric mean 2H7 1546 3E12 208 4B11 1098 5C6 1687 5H9 698 5F8 276 6B8 935 7D2 2255 8A5 1812 8C7 343 9A11 351 11D3 673 12C8 619 13D11 2125 13G3 739 14C2 423 Comparison 88

[0246] Example 4: Isotypes and cloning of VH / VL of anti-CD84 antibodies

[0247] To determine the Ig isotype of mouse antibodies, CD84-IgA1 protein was used as an immobilized antigen for incubation with hybridoma culture supernatants in ELISA. Antibody isotypes were detected using HRP-conjugated goat anti-mouse Igg1, Igg2a, Igg2b, Igg3, Igk, and Igl (SouthernBiotech) and developed using TMB substrate. All six antibodies were of g1 and kappa isotypes ( Figure 5). To clone VH / VL sequences from hybridomas, mRNA was extracted using the Quick-RNA Microprep kit (Zymo Research). The first-strand cDNA was synthesized using the SuperScript III First-Strand Synthesis System (Thermo Fisher Scientific). VH and VL fragments were amplified by PCR using a mouse Ig-primer set (Millipore Sigma) and OneTaq2X Master Mix (NEB). The amplified DNA fragments were purified using a DNA cleanup kit (Zymo Research) and ligated into a pGEM-T vector (Promega) for sequencing.

[0248] Example 5: CD84 is overexpressed in AML and is required for the growth of human AML cells Based on gene expression profiling datasets including large cohorts of AML patients (GSE 9476 and GSE13159), we observed that bone marrow mononuclear cells of AML samples showed significantly increased CD84 mRNA levels compared with those of normal healthy donors ( Fig.11A , left). Notably, myelomonocytic and monocytic leukemia patients with t(11q23) and inv(16) abnormalities showed even higher CD84 expression compared to other AML subtypes ( Fig.11A , highlighted in the right panel). We further confirmed that surface CD84 was overexpressed in AML cell lines (n=6) and primary cells (n=4) compared to that observed in healthy donors ( Fig. 11B In addition, we also found that higher CD84 expression was associated with shorter overall survival (OS) in AML patients (GSE10358) ( Fig. 11C ).

[0249] To investigate the role of CD84 in AML, both gain-of-function and loss-of-function studies were performed. Knockdown of CD84 by short hairpin RNA (shRNA) resulted in substantial inhibition of cell growth and apoptosis in AML cell lines ( Fig. 12A , left and right, respectively). In primary AML cells, we found that CD84 knockout significantly induced apoptosis and inhibited colony number ( Fig. 12B , left and right pictures respectively).

[0250] In addition, we knocked out CD84 in luciferase-expressing THP1 cells and then transplanted selected cells into immunodeficient NSG mice. Relative to control mice, attenuated tumor burden was observed in mice transplanted with CD84 knockout cells ( Fig. 12C , left) and prolonged survival ( Fig. 12C , right).

[0251] Example 6: Loss of CD84 expression impairs AML cell repopulation and AML maintenance

[0252] To understand the functional involvement of CD84 in leukemogenesis, we used a lentiviral vector-based shRNA system to knock down the expression of CD84 (shCD84-1 and shCD84-2) in mouse MLL-AF9-HSPC (hematopoietic stem / progenitor cells) pre-LSC (leukemic stem cells), which were generated by transducing mouse HSPC with lentivirus encoding the MLL-AF9 fusion oncogene. Fig.13A As shown in Figure 2, MLL-AF9-HSPC pre-LSCs transduced with shCD84 (CD84 KD) had slower cell growth ( Fig. 13B As expected, colony formation assay (CFA) indicated that stable CD84 KD with both shRNAs significantly reduced the number of colonies in transduced MLL-AF9-HSPC cells ( Fig. 13C To evaluate the role of CD84 in leukemogenesis in vivo, we performed mouse bone marrow transplantation experiments in sublethally irradiated C57BL / 6 syngeneic recipient mice. We found that mice transplanted with CD84KD MLL-AF9-HSPC cells had reduced bone marrow and spleen enlargement compared with recipient mice transplanted with MLL-AF9 cells transduced with sh-control vector ( FIG. 13D to FIG. 13E ).

[0253] We next performed in vivo transplantation (to evaluate the potential of CD84 in maintaining human AML). We transfected CD84 shRNA (sh-CD84-2) and control shRNA into primary AML patient cells. In this case, we also observed that recipient mice engrafted with primary CD84 knockout AML cells had significantly lower AML burden in the BM, spleen, and peripheral blood (PB) compared to mice engrafted with shcontrol AML cells ( Fig.14A We also found that spleen weight was reduced in CD84 knockout mice ( Fig. 14B ). Taken together, our data indicate that CD84 is required for the maintenance of AML.

[0254] Example 7: CD84 is a survival receptor in AML, and anti-CD84 antibodies strongly induce ADCC

[0255] To effectively target AML cells expressing CD84 in vivo, we generated three mouse anti-human chimeric monoclonal antibodies against CD84 (anti-CD84 antibodies), including monoclonal antibodies 5C6, 7D2, and 8A5. Antibody-dependent cellular cytotoxicity (ADCC) assays showed that anti-CD847D2 antibody induced higher cytotoxicity in THP1 cells with high CD84 expression, but not in MM1S cells with low CD84 expression ( Fig.15A ). We further investigated this using anti-CD84 7D2 antibody. As expected, anti-CD84 7D2 antibody strongly induced ADCC in THP1 cells in the presence of total peripheral blood mononuclear cells (PBMC) or purified NK cells as effector cells (E), which was dependent on the ratio of effector cells to target cells and the antibody dose ( Fig. 15B ). Notably, reduced tumor burden and prolonged survival were observed in luciferase-expressing THP1-xenografted NSG mice following anti-CD84 7D2 antibody treatment ( Fig.16A At the same time, we maintained the same treatment strategy in the U937-Luc+ xenograft mouse model. Compared with mice treated with IgG, mice treated with anti-CD84 antibody showed a significant reduction in disease burden by bioluminescence, accompanied by a significantly prolonged survival time ( Fig. 16B Consistently, in AML patient-derived xenograft (PDX) models, we found reduced engraftment (indicated by human CD45 / CD33) in the bone marrow, spleen, and peripheral blood, as well as reduced splenomegaly ( ) after three weeks of anti-CD84 antibody treatment. Fig.17 ). In conclusion, our results indicate that CD84 is a survival receptor in AML and that anti-CD84 antibodies are promising therapy for AML patients.

[0256] Example 8: CD84 and 8A5 interface

[0257] The buried surface area of ​​the CD84 / 8A5 structure was calculated using the PISA (Protein, Interface, Structure and Assembly) tool. The interface area between the human heavy chain (HC) and CD84 (in , calculated as the difference in the total accessible surface area of ​​the separated structure and the interface structure divided by 2) There are many hydrogen bonds within the interface, including Ser99(HC)-Gln24(CD84), Thr53(HC)-Gln24(CD84), Ser33(HC)–Glu25(CD84); Thr30(HC)-Gln24(CD84) and the salt bridge His25(HC)Glu25(CD84).

[0258] The list of CD84 residues buried at the HC-CD84 interface is Val21, Asn22, ILe23, Gln24, Glu25, Pro26, Arg27, Gln28, Ser48, Glu49 Pro70, Tyr72, Ala95, Asp96, Pro97, Tyr98, Thr100, Lys102

[0259] The list of 8A5 residues of HC is Thr30, Thr31, Tyr32, Ser33, His35, Trp47, Trp50, Asp52, Thr53, Ala54, Thr55, Glu57, Pro58, Thr59, Tyr60, Ala61, Lys65, Ser99, Tyr101, Trp102, Tyr103, Phe104

[0260] The interface between human light chain (LC) and CD84 is There are many hydrogen bonds within this interface, including Asn 33(LC)-Ser48(CD84), Lys55(LC)–Glu49(CD84), Gly96(LC)-Arg27(CD84), and Asn35(LC)–Ser48(CD84), and salt bridges Lys55(LC)-Glu49(CD84) and Glu39(LC)-Arg27(CD84).

[0261] The list of CD84 residues buried at the LC-CD84 interface is Glu25, Arg27, Gln28, Val29, Lys30, Pro45, Gly46, Ssp47, Ser48, Glu49, Gln94 and Ala95

[0262] The list of 8A5 residues of LC is His31, Ile32, Asn33, Gly34, Asn35, Tyr37, Glu39, Lys55, Phe94, Gly96, Ser97, His98, Vla99, Trp101

[0263] Informal Sequence Listing

[0264]

[0265]

[0266]

[0267]

[0268]

[0269]

[0270] Implementation Plan

[0271] P Embodiment 1. An anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:75, CDR H2 as shown in SEQ ID NO:76, and CDR H3 as shown in SEQ ID NO:77; and wherein the light chain variable domain comprises CDR L1 as shown in SEQ ID NO:78, CDR L2 as shown in SEQ ID NO:79, and CDR L3 as shown in SEQ ID NO:80.

[0272] P embodiment 2. An anti-CD84 antibody according to P embodiment 1, wherein the anti-CD84 antibody is a chimeric antibody.

[0273] P Embodiment 3. An anti-CD84 antibody according to P Embodiment 1 or 2, wherein the heavy chain variable domain comprises FR H1 as shown in SEQ ID NO:81, FR H2 as shown in SEQ ID NO:82, FR H3 as shown in SEQ ID NO:83 and FR H4 as shown in SEQ ID NO:84.

[0274] P Embodiment 4. An anti-CD84 antibody according to any one of P Embodiments 1 to 3, wherein the light chain variable domain comprises FR L1 as shown in SEQ ID NO:85, FR L2 as shown in SEQ ID NO:86, FR L3 as shown in SEQ ID NO:87, and FR L4 as shown in SEQ ID NO:88.

[0275] P Embodiment 5. An anti-CD84 antibody according to any one of P Embodiments 1 to 4, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO: 89.

[0276] P Embodiment 6. An anti-CD84 antibody according to any one of P Embodiments 1 to 5, wherein the light chain variable domain comprises the sequence of SEQ ID NO:90.

[0277] P Embodiment 7. An anti-CD84 antibody according to any one of P Embodiments 1 to 6, wherein the anti-CD84 antibody comprises a heavy chain comprising the sequence of SEQ ID NO:91.

[0278] P Embodiment 8. An anti-CD84 antibody according to any one of P Embodiments 1 to 7, wherein the anti-CD84 antibody comprises a light chain comprising the sequence of SEQ ID NO:92.

[0279] P Embodiment 9. An anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:19, CDR H2 as shown in SEQ ID NO:20, and CDR H3 as shown in SEQ ID NO:21; and wherein the light chain variable domain comprises CDR L1 as shown in SEQ ID NO:22, CDR L2 as shown in SEQ ID NO:23, and CDR L3 as shown in SEQ ID NO:24.

[0280] P Embodiment 10. An anti-CD84 antibody according to P Embodiment 9, wherein the anti-CD84 antibody is a humanized antibody.

[0281] P Embodiment 11. An anti-CD84 antibody according to any one of P Embodiments 9 to 10, wherein the antibody comprises a light chain comprising the sequence of SEQ ID NO: 37.

[0282] P Embodiment 12. An anti-CD84 antibody according to any one of P Embodiments 9 to 11, wherein the antibody comprises a heavy chain comprising the sequence of SEQ ID NO: 38

[0283] P Embodiment 13. An anti-CD84 antibody according to P Embodiment 9, wherein the anti-CD84 antibody is a chimeric antibody.

[0284] P Embodiment 14. An anti-CD84 antibody according to P Embodiment 9 or 13, wherein the heavy chain variable domain comprises FR H1 as shown in SEQ ID NO:25, FR H2 as shown in SEQ ID NO:26, FR H3 as shown in SEQ ID NO:27 and FR H4 as shown in SEQ ID NO:28.

[0285] P Embodiment 15. An anti-CD84 antibody according to any one of P Embodiments 9 or 13 to 14, wherein the light chain variable domain comprises FR L1 as shown in SEQ ID NO:29, FR L2 as shown in SEQ ID NO:30, FR L3 as shown in SEQ ID NO:31 and FR L4 as shown in SEQ ID NO:32.

[0286] P Embodiment 16. An anti-CD84 antibody according to any one of P Embodiments 9 or 13 to 15, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO: 33.

[0287] P Embodiment 17. An anti-CD84 antibody according to any one of P Embodiments 9 or 13 to 16, wherein the light chain variable domain comprises the sequence of SEQ ID NO: 34.

[0288] P Embodiment 18. An anti-CD84 antibody according to any one of P Embodiments 9 or 13 to 17, wherein the anti-CD84 antibody comprises a heavy chain comprising the sequence of SEQ ID NO: 35.

[0289] P Embodiment 19. An anti-CD84 antibody according to any one of P Embodiments 9 or 13 to 18, wherein the anti-CD84 antibody comprises a light chain comprising the sequence of SEQ ID NO: 36.

[0290] P Embodiment 20. An anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:39, CDR H2 as shown in SEQ ID NO:40, and CDR H3 as shown in SEQ ID NO:41; and wherein the light chain variable domain comprises CDR L1 as shown in SEQ ID NO:42, CDR L2 as shown in SEQ ID NO:43, and CDR L3 as shown in SEQ ID NO:44.

[0291] P embodiment 21. An anti-CD84 antibody according to P embodiment 20, wherein the anti-CD84 antibody is a chimeric antibody.

[0292] P Embodiment 22. An anti-CD84 antibody according to P Embodiment 20 or 21, wherein the heavy chain variable domain comprises FR H1 as shown in SEQ ID NO:45, FR H2 as shown in SEQ ID NO:46, FR H3 as shown in SEQ ID NO:47 and FR H4 as shown in SEQ ID NO:48.

[0293] P Embodiment 23. An anti-CD84 antibody according to any one of P Embodiments 20 to 22, wherein the light chain variable domain comprises FR L1 as shown in SEQ ID NO:49, FR L2 as shown in SEQ ID NO:50, FR L3 as shown in SEQ ID NO:51, and FR L4 as shown in SEQ ID NO:52.

[0294] P Embodiment 24. An anti-CD84 antibody according to any one of P Embodiments 20 to 23, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO: 53.

[0295] P Embodiment 25. An anti-CD84 antibody according to any one of P Embodiments 20 to 24, wherein the light chain variable domain comprises the sequence of SEQ ID NO: 54.

[0296] P Embodiment 26. An anti-CD84 antibody according to any one of P Embodiments 20 to 25, wherein the anti-CD84 antibody comprises a heavy chain comprising the sequence of SEQ ID NO: 55.

[0297] P Embodiment 27. An anti-CD84 antibody according to any one of P Embodiments 20 to 26, wherein the anti-CD84 antibody comprises a light chain comprising the sequence of SEQ ID NO: 56.

[0298] P Embodiment 28. An anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:57, CDR H2 as shown in SEQ ID NO:58, and CDR H3 as shown in SEQ ID NO:59; and wherein the light chain variable domain comprises CDR L1 as shown in SEQ ID NO:60, CDR L2 as shown in SEQ ID NO:61, and CDR L3 as shown in SEQ ID NO:62.

[0299] P Embodiment 29. An anti-CD84 antibody according to P Embodiment 28, wherein the anti-CD84 antibody is a chimeric antibody.

[0300] P Embodiment 30. An anti-CD84 antibody according to P Embodiment 28 or 29, wherein the heavy chain variable domain comprises FR H1 as shown in SEQ ID NO:63, FR H2 as shown in SEQ ID NO:64, FR H3 as shown in SEQ ID NO:65 and FR H4 as shown in SEQ ID NO:66.

[0301] P Embodiment 31. An anti-CD84 antibody according to any one of P Embodiments 28 to 30, wherein the light chain variable domain comprises FR L1 as shown in SEQ ID NO:67, FR L2 as shown in SEQ ID NO:68, FR L3 as shown in SEQ ID NO:69 and FR L4 as shown in SEQ ID NO:70.

[0302] P Embodiment 32. An anti-CD84 antibody according to any one of P Embodiments 28 to 31, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO: 71.

[0303] P Embodiment 33. An anti-CD84 antibody according to any one of P Embodiments 28 to 32, wherein the light chain variable domain comprises the sequence of SEQ ID NO: 72.

[0304] P Embodiment 34. An anti-CD84 antibody according to any one of P Embodiments 28 to 33, wherein the anti-CD84 antibody comprises a heavy chain comprising the sequence of SEQ ID NO: 73.

[0305] P Embodiment 35. An anti-CD84 antibody according to any one of P Embodiments 28 to 34, wherein the anti-CD84 antibody comprises a light chain comprising the sequence of SEQ ID NO: 74.

[0306] P Embodiment 36. An anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:93, CDR H2 as shown in SEQ ID NO:94, and CDR H3 as shown in SEQ ID NO:95; and wherein the light chain variable domain comprises CDR L1 as shown in SEQ ID NO:96, CDR L2 as shown in SEQ ID NO:97, and CDR L3 as shown in SEQ ID NO:98.

[0307] P Embodiment 37. An anti-CD84 antibody according to P Embodiment 36, wherein the anti-CD84 antibody is a chimeric antibody.

[0308] P Embodiment 38. An anti-CD84 antibody according to P Embodiment 36 or 37, wherein the heavy chain variable domain comprises FR H1 as shown in SEQ ID NO:99, FR H2 as shown in SEQ ID NO:100, FR H3 as shown in SEQ ID NO:101 and FR H4 as shown in SEQ ID NO:102.

[0309] P Embodiment 39. An anti-CD84 antibody according to any one of P Embodiments 36 to 38, wherein the light chain variable domain comprises FR L1 as shown in SEQ ID NO:103, FR L2 as shown in SEQ ID NO:104, FR L3 as shown in SEQ ID NO:105 and FR L4 as shown in SEQ ID NO:106.

[0310] P Embodiment 40. An anti-CD84 antibody according to any one of P Embodiments 36 to 39, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO: 107.

[0311] P Embodiment 41. An anti-CD84 antibody according to any one of P Embodiments 36 to 40, wherein the light chain variable domain comprises the sequence of SEQ ID NO: 108.

[0312] P Embodiment 42. An anti-CD84 antibody according to any one of P Embodiments 36 to 41, wherein the anti-CD84 antibody comprises a heavy chain comprising the sequence of SEQ ID NO: 109.

[0313] P Embodiment 43. An anti-CD84 antibody according to any one of P Embodiments 36 to 42, wherein the anti-CD84 antibody comprises a light chain comprising the sequence of SEQ ID NO:110.

[0314] P Embodiment 44. An anti-CD84 antibody according to any one of P Embodiments 1 to 43, wherein the anti-CD84 antibody is a Fab' fragment.

[0315] P Embodiment 45. An anti-CD84 antibody according to any one of P Embodiments 1 to 44, wherein the anti-CD84 antibody is IgG.

[0316] P Embodiment 46. An anti-CD84 antibody according to any one of P Embodiments 1, 3 to 6, 9, 14 to 17, 20, 22 to 25, 28, 30 to 33 or 36 or 38 to 41, wherein the light chain variable domain and the heavy chain variable domain form part of a scFv.

[0317] P Embodiment 47. An anti-CD84 antibody according to any one of P Embodiments 1 to 46, wherein the anti-CD84 antibody is capable of binding to CD84 protein.

[0318] P Embodiment 48. An anti-CD84 antibody according to any one of P Embodiments 1 to 47, wherein the anti-CD84 antibody binds to the CD84 protein.

[0319] P Embodiment 49. An anti-CD84 antibody according to P Embodiment 47, wherein the CD84 protein is a human CD84 protein.

[0320] P Embodiment 50. An anti-CD84 antibody according to P Embodiment 48 or 49, wherein the CD84 protein forms part of a cell.

[0321] P Embodiment 51. An anti-CD84 antibody according to any one of P Embodiments 47 to 50, wherein the CD84 protein is expressed on the surface of a cell.

[0322] P Embodiment 52. An anti-CD84 antibody according to P Embodiment 50 or 51, wherein the cell is a cancer cell.

[0323] P Embodiment 53. An anti-CD84 antibody according to P Embodiment 52, wherein the cancer cell is a leukemia cancer cell, a myeloid cancer cell, or a lymphoma cancer cell.

[0324] P Embodiment 54. An anti-CD84 antibody, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:19, CDR H2 as shown in SEQ ID NO:20, and CDR H3 as shown in SEQ ID NO:21, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:22, CDR L2 as shown in SEQ ID NO:23, and CDR L3 as shown in SEQ ID NO:24.

[0325] P Embodiment 55. An anti-CD84 antibody, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:39, CDR H2 as shown in SEQ ID NO:40, and CDR H3 as shown in SEQ ID NO:41, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:42, CDR L2 as shown in SEQ ID NO:43, and CDR L3 as shown in SEQ ID NO:44.

[0326] P Embodiment 56. An anti-CD84 antibody, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:57, CDR H2 as shown in SEQ ID NO:58, and CDR H3 as shown in SEQ ID NO:59, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:60, CDR L2 as shown in SEQ ID NO:61, and CDR L3 as shown in SEQ ID NO:62.

[0327] P Embodiment 57. An anti-CD84 antibody, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:75, CDR H2 as shown in SEQ ID NO:76, and CDR H3 as shown in SEQ ID NO:77, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:78, CDR L2 as shown in SEQ ID NO:79, and CDR L3 as shown in SEQ ID NO:80.

[0328] P Embodiment 58. An anti-CD84 antibody, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:93, CDR H2 as shown in SEQ ID NO:94, and CDR H3 as shown in SEQ ID NO:95, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:96, CDR L2 as shown in SEQ ID NO:97, and CDR L3 as shown in SEQ ID NO:98.

[0329] P Embodiment 59. An anti-CD84 antibody according to any one of P Embodiments 1 to 58, wherein the anti-CD84 antibody is linked to a therapeutic or diagnostic moiety.

[0330] P Embodiment 60. An anti-CD84 antibody according to P Embodiment 59, wherein the therapeutic moiety is an anti-cancer moiety.

[0331] P embodiment 61. An isolated nucleic acid encoding an anti-CD84 antibody according to any one of P embodiments 1 to 60.

[0332] 62. A cell comprising the anti-CD84 antibody of any one of claims 1 to 60 or the nucleic acid of embodiment P 61.

[0333] P Embodiment 63. A pharmaceutical composition comprising a therapeutically effective amount of an antibody according to any one of P Embodiments 1 to 60 and a pharmaceutically acceptable excipient.

[0334] P Embodiment 64. A method for forming an antibody capable of binding to CD84, the method comprising immunizing a mammal with a peptide comprising the sequence of SEQ ID NO:111.

[0335] P Embodiment 65. A method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of an anti-CD84 antibody according to any one of P Embodiments 1 to 60, or a pharmaceutical composition according to P Embodiment 63, thereby treating the cancer in the subject.

[0336] P Embodiment 66. A method for treating cancer according to P Embodiment 65, wherein the cancer is lymphoma, myeloma or leukemia.

[0337] P Embodiment 67. A method for treating cancer according to P Embodiment 65 or 66, wherein the cancer is acute myeloid leukemia (AML).

Claims

1. An anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO: 75, CDR H2 as shown in SEQ ID NO: 76, and CDR H3 as shown in SEQ ID NO: 77; and The light chain variable domain comprises CDR L1 as shown in SEQ ID NO:78, CDR L2 as shown in SEQ ID NO:79 and CDR L3 as shown in SEQ ID NO:

80.

2. The anti-CD84 antibody of claim 1, wherein the anti-CD84 antibody is a chimeric antibody.

3. The anti-CD84 antibody of claim 1, wherein the heavy chain variable domain comprises FR H1 as shown in SEQ ID NO: 81, FR H2 as shown in SEQ ID NO: 82, FR H3 as shown in SEQ ID NO: 83, and FR H4 as shown in SEQ ID NO:

84.

4. The anti-CD84 antibody of claim 1, wherein the light chain variable domain comprises FR L1 as shown in SEQ ID NO: 85, FR L2 as shown in SEQ ID NO: 86, FR L3 as shown in SEQ ID NO: 87, and FR L4 as shown in SEQ ID NO:

88.

5. The anti-CD84 antibody of claim 1, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO:

89.

6. The anti-CD84 antibody of claim 1, wherein the light chain variable domain comprises the sequence of SEQ ID NO:

90.

7. The anti-CD84 antibody of claim 1, wherein the anti-CD84 antibody comprises a heavy chain comprising the sequence of SEQ ID NO:

91.

8. The anti-CD84 antibody of claim 1, wherein the anti-CD84 antibody comprises a light chain comprising the sequence of SEQ ID NO:

92.

9. An anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO: 19, CDR H2 as shown in SEQ ID NO: 20, and CDR H3 as shown in SEQ ID NO: 21; and The light chain variable domain comprises CDR L1 as shown in SEQ ID NO:22, CDR L2 as shown in SEQ ID NO:23 and CDR L3 as shown in SEQ ID NO:

24.

10. The anti-CD84 antibody of claim 9, wherein the anti-CD84 antibody is a humanized antibody.

11. The anti-CD84 antibody of claim 9, wherein the antibody comprises a light chain comprising the sequence of SEQ ID NO:

37.

12. The anti-CD84 antibody of claim 9, wherein the antibody comprises a heavy chain comprising the sequence of SEQ ID NO:

38.

13. The anti-CD84 antibody of claim 9, wherein the anti-CD84 antibody is a chimeric antibody.

14. The anti-CD84 antibody of claim 9, wherein the heavy chain variable domain comprises FR H1 as shown in SEQ ID NO: 25, FR H2 as shown in SEQ ID NO: 26, FR H3 as shown in SEQ ID NO: 27, and FR H4 as shown in SEQ ID NO:

28.

15. The anti-CD84 antibody of claim 9, wherein the light chain variable domain comprises FR L1 as shown in SEQ ID NO: 29, FR L2 as shown in SEQ ID NO: 30, FR L3 as shown in SEQ ID NO: 31, and FR L4 as shown in SEQ ID NO:

32.

16. The anti-CD84 antibody of claim 9, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO:

33.

17. The anti-CD84 antibody of claim 9, wherein the light chain variable domain comprises the sequence of SEQ ID NO:

34.

18. The anti-CD84 antibody of claim 9, wherein the anti-CD84 antibody comprises a heavy chain comprising the sequence of SEQ ID NO:

35.

19. The anti-CD84 antibody of claim 9, wherein the anti-CD84 antibody comprises a light chain comprising the sequence of SEQ ID NO:

36.

20. An anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO: 39, CDR H2 as shown in SEQ ID NO: 40, and CDR H3 as shown in SEQ ID NO: 41; and The light chain variable domain comprises CDR L1 as shown in SEQ ID NO:42, CDR L2 as shown in SEQ ID NO:43 and CDR L3 as shown in SEQ ID NO:

44.

21. The anti-CD84 antibody of claim 20, wherein the anti-CD84 antibody is a chimeric antibody.

22. The anti-CD84 antibody of claim 20, wherein the heavy chain variable domain comprises FR H1 as shown in SEQ ID NO:45, FR H2 as shown in SEQ ID NO:46, FR H3 as shown in SEQ ID NO:47, and FR H4 as shown in SEQ ID NO:

48.

23. The anti-CD84 antibody of claim 20, wherein the light chain variable domain comprises FR L1 as shown in SEQ ID NO:49, FR L2 as shown in SEQ ID NO:50, FR L3 as shown in SEQ ID NO:51, and FR L4 as shown in SEQ ID NO:

52.

24. The anti-CD84 antibody of claim 20, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO:

53.

25. The anti-CD84 antibody of claim 20, wherein the light chain variable domain comprises the sequence of SEQ ID NO:

54.

26. The anti-CD84 antibody of claim 20, wherein the anti-CD84 antibody comprises a heavy chain comprising the sequence of SEQ ID NO:

55.

27. The anti-CD84 antibody of claim 20, wherein the anti-CD84 antibody comprises a light chain comprising the sequence of SEQ ID NO:

56.

28. An anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO: 57, CDR H2 as shown in SEQ ID NO: 58, and CDR H3 as shown in SEQ ID NO: 59; and The light chain variable domain comprises CDR L1 as shown in SEQ ID NO:60, CDR L2 as shown in SEQ ID NO:61 and CDR L3 as shown in SEQ ID NO:

62.

29. The anti-CD84 antibody of claim 28, wherein the anti-CD84 antibody is a chimeric antibody.

30. The anti-CD84 antibody of claim 28, wherein the heavy chain variable domain comprises FR H1 as shown in SEQ ID NO: 63, FR H2 as shown in SEQ ID NO: 64, FR H3 as shown in SEQ ID NO: 65, and FR H4 as shown in SEQ ID NO:

66.

31. The anti-CD84 antibody of claim 28, wherein the light chain variable domain comprises FR L1 as shown in SEQ ID NO: 67, FR L2 as shown in SEQ ID NO: 68, FR L3 as shown in SEQ ID NO: 69, and FR L4 as shown in SEQ ID NO:

70.

32. The anti-CD84 antibody of claim 28, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO:

71.

33. The anti-CD84 antibody of claim 28, wherein the light chain variable domain comprises the sequence of SEQ ID NO:

72.

34. The anti-CD84 antibody of claim 28, wherein the anti-CD84 antibody comprises a heavy chain comprising the sequence of SEQ ID NO:

73.

35. The anti-CD84 antibody of claim 28, wherein the anti-CD84 antibody comprises a light chain comprising the sequence of SEQ ID NO:

74.

36. An anti-CD84 antibody comprising a light chain variable domain and a heavy chain variable domain, wherein the heavy chain variable domain comprises CDR H1 as shown in SEQ ID NO:93, CDR H2 as shown in SEQ ID NO:94, and CDR H3 as shown in SEQ ID NO:95; and The light chain variable domain comprises CDR L1 as shown in SEQ ID NO:96, CDR L2 as shown in SEQ ID NO:97 and CDR L3 as shown in SEQ ID NO:

98.

37. The anti-CD84 antibody of claim 36, wherein the anti-CD84 antibody is a chimeric antibody.

38. The anti-CD84 antibody of claim 36, wherein the heavy chain variable domain comprises FR H1 as shown in SEQ ID NO:99, FR H2 as shown in SEQ ID NO:100, FR H3 as shown in SEQ ID NO:101, and FR H4 as shown in SEQ ID NO:

102.

39. The anti-CD84 antibody of claim 36, wherein the light chain variable domain comprises FR L1 as shown in SEQ ID NO: 103, FR L2 as shown in SEQ ID NO: 104, FR L3 as shown in SEQ ID NO: 105, and FR L4 as shown in SEQ ID NO:

106.

40. The anti-CD84 antibody of claim 36, wherein the heavy chain variable domain comprises the sequence of SEQ ID NO:

107.

41. The anti-CD84 antibody of claim 36, wherein the light chain variable domain comprises the sequence of SEQ ID NO:

108.

42. The anti-CD84 antibody of claim 36, wherein the anti-CD84 antibody comprises a heavy chain comprising the sequence of SEQ ID NO:

109.

43. The anti-CD84 antibody of claim 36, wherein the anti-CD84 antibody comprises a light chain comprising the sequence of SEQ ID NO:

110.

44. The anti-CD84 antibody of any one of claims 1, 9, 20, 28 or 36, wherein the anti-CD84 antibody is a Fab' fragment.

45. The anti-CD84 antibody of any one of claims 1, 9, 20, 28 or 36, wherein the anti-CD84 antibody is an IgG.

46. ​​An anti-CD84 antibody according to any one of claims 1, 9, 20, 28 or 36, wherein the light chain variable domain and the heavy chain variable domain form part of a scFv.

47. The anti-CD84 antibody of any one of claims 1, 9, 20, 28 or 36, wherein the anti-CD84 antibody is capable of binding to CD84 protein.

48. The anti-CD84 antibody of any one of claims 1, 9, 20, 28 or 36, wherein the anti-CD84 antibody binds to a CD84 protein.

49. The anti-CD84 antibody of claim 47, wherein the CD84 protein is a human CD84 protein.

50. The anti-CD84 antibody of claim 48, wherein the CD84 protein forms part of a cell.

51. The anti-CD84 antibody of claim 47, wherein the CD84 protein is expressed on the surface of a cell.

52. The anti-CD84 antibody of claim 50, wherein the cell is a cancer cell.

53. The anti-CD84 antibody of claim 52, wherein the cancer cell is a leukemia cancer cell, a myeloid cancer cell, or a lymphoma cancer cell.

54. An anti-CD84 antibody, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO: 19, CDR H2 as shown in SEQ ID NO: 20, and CDR H3 as shown in SEQ ID NO: 21, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO: 22, CDR L2 as shown in SEQ ID NO: 23, and CDR L3 as shown in SEQ ID NO:

24.

55. An anti-CD84 antibody, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:39, CDR H2 as shown in SEQ ID NO:40, and CDR H3 as shown in SEQ ID NO:41, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:42, CDR L2 as shown in SEQ ID NO:43, and CDR L3 as shown in SEQ ID NO:

44.

56. An anti-CD84 antibody, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:57, CDR H2 as shown in SEQ ID NO:58, and CDR H3 as shown in SEQ ID NO:59, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:60, CDR L2 as shown in SEQ ID NO:61, and CDR L3 as shown in SEQ ID NO:

62.

57. An anti-CD84 antibody, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:75, CDR H2 as shown in SEQ ID NO:76, and CDR H3 as shown in SEQ ID NO:77, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:78, CDR L2 as shown in SEQ ID NO:79, and CDR L3 as shown in SEQ ID NO:

80.

58. An anti-CD84 antibody, wherein the anti-CD84 antibody binds to the same epitope as an antibody comprising: a heavy chain variable domain comprising CDR H1 as shown in SEQ ID NO:93, CDR H2 as shown in SEQ ID NO:94, and CDR H3 as shown in SEQ ID NO:95, and a light chain variable domain comprising CDR L1 as shown in SEQ ID NO:96, CDR L2 as shown in SEQ ID NO:97, and CDR L3 as shown in SEQ ID NO:

98.

59. The anti-CD84 antibody of any one of claims 1, 9, 20, 28, 36, 54, 55, 56, 57 or 58, wherein the anti-CD84 antibody is linked to a therapeutic or diagnostic moiety.

60. The anti-CD84 antibody of claim 59, wherein the therapeutic moiety is an anti-cancer moiety.

61. An isolated nucleic acid encoding the anti-CD84 antibody of any one of claims 1, 9, 20, 28, 36, 54, 55, 56, 57 or 58.

62. A cell comprising the anti-CD84 antibody of any one of claims 1, 9, 20, 28, 36, 54, 55, 56, 57 or 58 or the nucleic acid of claim 61.

63. A pharmaceutical composition comprising a therapeutically effective amount of an antibody according to any one of claims 1, 9, 20, 28, 36, 54, 55, 56, 57 or 58 and a pharmaceutically acceptable excipient.

64. A method for generating an antibody capable of binding to CD84, the method comprising immunizing a mammal with a peptide comprising the sequence of SEQ ID NO:

111.

65. A method for treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of an anti-CD84 antibody according to any one of claims 1, 9, 20, 28, 36, 54, 55, 56, 57 or 58, or a pharmaceutical composition according to claim 63, thereby treating the cancer in the subject.

66. The method of treating cancer according to claim 65, wherein the cancer is lymphoma, myeloma or leukemia.

67. The method of treating cancer according to claim 65, wherein the cancer is acute myeloid leukemia (AML).

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