Conjugated chemical inducers of degradation, and methods of use

Abstract

To provide antibody-CIDE conjugates (Ab-CIDEs), pharmaceutical compositions containing them, and their use in treating diseases and conditions where targeted protein degradation is beneficial.SOLUTION: For example, antibody-CIDE conjugates of the following formula are illustrated.SELECTED DRAWING: None

Description

Technical field

[0001] Cross-reference to related applications This application claims priority to and benefit from U.S. Provisional Patent Application No. 62 / 749,812, filed October 24, 2018, the disclosure of which is hereby incorporated by reference in its entirety.

[0002] Sequence listing references submitted as text files via EFS-WEB An official copy of the Sequence Listing will be submitted electronically via EFS-Web as a Sequence Listing in ASCII format with a file named 515668_SEQLIST.TXT having a size of 274 kilobytes, created on October 24, 2018, and submitted concurrently with the specification. The Sequence Listing contained in this ASCII document is part of the specification and is hereby incorporated by reference in its entirety.

[0003] The subject matter described herein generally relates to degradant conjugates comprising antibody-proteolytic targeting chimeric molecules useful for promoting intracellular degradation of target proteins. [Background technology]

[0004] Cell maintenance and normal function require the controlled breakdown of cellular proteins. For example, degradation of regulatory proteins triggers events in the cell cycle such as DNA replication and chromosome segregation. Degradation of such proteins therefore affects cell proliferation, differentiation and death.

[0005] Inhibitors of proteins can block or reduce intracellular protein activity, but intracellular proteolysis can also reduce activity or completely eliminate target proteins. Thus, exploiting the cell's proteolytic pathways can provide a means to reduce or eliminate protein activity. One of the cell's major degradation pathways is known as the ubiquitin-proteasome system. In this system, proteins are marked for degradation by the proteasome by ubiquitylating the proteins. Ubiquitination of proteins is accomplished by E3 ubiquitin ligases that bind to proteins and add ubiquitin molecules to proteins. E3 ubiquitin ligases are part of a pathway that includes E1 and E2 ubiquitin ligases, which make ubiquitin available to E3 ubiquitin ligases to add to proteins.

[0006] To exploit this degradation pathway, molecular constructs combine an E3 ubiquitin ligase with a protein targeted for degradation and a targeting antibody. To promote proteins for degradation by the proteasome, the molecular construct consists of a group that binds the E3 ubiquitin ligase and a group that binds the protein target for degradation. These groups are typically connected by a linker. This molecular construct can bring the E3 ubiquitin ligase into close proximity to the protein so that the E3 ubiquitin ligase is ubiquitinated and labeled for degradation. However, the relatively large size of molecular constructs can be problematic for targeted delivery.

[0007] There is a continuing need in the art for enhanced targeted delivery of such molecular constructs to cells containing protein targets. The subject matter described herein addresses this and other deficiencies in the art. [Outline of the invention]

[0008] In one aspect, the subject matter described herein is a covalently linked Ab-CIDE (PAC), wherein the covalent bond positions connecting the components of the Ab-CIDE: Ab, L1 (linker 1), L2 (linker 2), the protein binding group and the E3 ligase binding group can be tailored as desired to prepare Ab-CIDEs with desired properties such as in vivo pharmacokinetics, stability and solubility.

[0009] In one aspect, the subject matter described herein has the formula: Ab-(L1-D) p During the ceremony, D is a CIDE with structure E3LB-L2-PB; E3LB is covalently attached to L2, said E3LB being a group that binds an E3 ligase, said E3 ligase being von Hippel-Gendau (VHL); L2 is a linker covalently attached to E3LB and PB; PB is a protein binding group covalently attached to L2, said PB is a group that binds BRD4 or ERα, including all variants, mutations, splice variants, indels and fusions thereof; Ab is an antibody covalently attached to L1; L1 is a linker covalently attached to Ab and D; and p has a value of about 1 to about 8; Conjugated chemical degradation inducers (“CIDE”) having

[0010] Another aspect of the subject matter described herein is a pharmaceutical composition comprising Ab-CIDE and one or more pharmaceutically acceptable excipients.

[0011] Another aspect of the subject matter described herein is the use of Ab-CIDE in methods of treating conditions and diseases by administering to a subject a pharmaceutical composition comprising Ab-CIDE.

[0012] Another aspect of the subject matter described herein is a method of making Ab-CIDE.

[0013] Another aspect of the subject matter described herein is an article of manufacture comprising a pharmaceutical composition comprising an Ab-CIDE, a container, and an insert or label indicating that the pharmaceutical composition can be used to treat a disease or condition. [Brief description of the drawing]

[0014]

Figure 1

Figure 2

[0015] Disclosed herein are antibody-chemical degradation inducer (“CIDE”) conjugates (referred to herein as Ab-CIDE or PAC) useful for targeted proteolysis and the treatment of related diseases and disorders. The subject matter described herein utilizes antibody targeting to direct CIDE to target cells or tissues. As described herein, linking an antibody to CIDE to form Ab-CIDE has been shown to deliver CIDE to target cells or tissues. As shown herein, for example, in the Examples, cells expressing antigen can be targeted by antigen-specific Ab-CIDE, whereby the CIDE portion of Ab-CIDE is delivered into target cells. CIDE containing antibodies to antigens not found on cells does not result in significant intracellular delivery of CIDE to cells.

[0016] Accordingly, the subject matter described herein relates to Ab-CIDE compositions that lead to ubiquitination of target proteins and subsequent protein degradation. The composition comprises an antibody covalently attached to a linker (L1), which is covalently attached at any available point of attachment to CIDE, the CIDE comprising an E3 ubiquitin ligase binding (E3LB) moiety that recognizes an E3 ubiquitin ligase protein that is VHL or XIAP and a protein binding moiety (PB) that recognizes a target protein that is ERα or BRD4. The subject matter described herein is useful for modulating protein activity and treating diseases and conditions associated with protein activity.

[0017] The subject matter disclosed herein will now be described more fully below. However, many modifications and other embodiments of the subject matter disclosed herein will come to mind to one skilled in the art to which the subject matter disclosed herein pertains having the benefit of the teachings presented in the foregoing description. Accordingly, the subject matter disclosed herein should not be limited to the particular embodiments disclosed, and modifications and other embodiments are intended to be included within the scope of the appended claims. In other words, the subject matter described herein encompasses all alternatives, modifications and equivalents. In the event that one or more of the incorporated literature, patents, and similar material differs from or conflicts with this application (including but not limited to defined terms, usage of terms, techniques described, etc.), this application will control. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety.

[0018] I. Definition The term "CIDE" generally refers to a proteolytically targeted chimeric molecule having three components, an E3 ubiquitin ligase linking group (E3LB), a linker L2, and a protein linking group (PB).

[0019] The term "residue," "moiety," or "group" refers to a moiety that is covalently bonded or linked to another moiety. As an example, a residue of a compound has an atom(s) of the compound, such as hydrogen or hydroxy, covalently replaced thereby linking the residue to another component of CIDE, L1-CIDE or Ab-CIDE. For example, "a residue of CIDE" refers to CIDE covalently attached to one or more groups such as linker L2, which itself can optionally be further linked to an antibody.

[0020] The terms "covalently bound" or "covalently linked" refer to chemical bonds formed by the sharing of one or more pairs of electrons.

[0021] The term "peptidomimetic" or PM as used herein means a non-peptide chemical moiety. Peptides are short chains of amino acid monomers linked by peptide (amide) bonds, which are covalent chemical bonds formed when the carboxyl group of one amino acid reacts with the amino group of another amino acid. The shortest peptides are dipeptides consisting of two amino acids joined by a single peptide bond, followed by tripeptides, tetrapeptides, and so on. Peptidomimetic chemical moieties include non-amino acid chemical moieties. A peptidomimetic chemical moiety can also comprise one or more amino acids separated by one or more non-amino acid chemical units. A peptidomimetic chemical moiety does not contain two or more contiguous amino acids linked by peptide bonds in any part of its chemical structure.

[0022] The term "antibody" herein is used in the broadest sense and specifically covers monoclonal antibodies, polyclonal antibodies, dimers, multimers, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, so long as they exhibit the desired biological activity (Miller et al (2003) Jour. of Immunology 170:4854-4861). Antibodies may be murine, human, humanized, chimeric, or derived from other species. Antibodies are proteins produced by the immune system that are capable of recognizing and binding to specific antigens (Janeway, C., Travers, P., Walport, M., Shlomchik (2001) Immuno Biology, 5th Ed., Garland Publishing, New York). A target antigen generally has numerous binding sites, also called epitopes, recognized by the CDRs (complementarity determining regions) of multiple antibodies. Each antibody that specifically binds a different epitope has a different structure. Thus, one antigen can have more than one corresponding antibody. Antibodies include full-length immunoglobulin molecules or immunologically active portions of full-length immunoglobulin molecules, i.e., molecules that contain an antigen-binding site that immunospecifically binds an antigen of a target or portion thereof of interest, including, but not limited to, cancer cells or cells that produce autoimmune antibodies associated with autoimmune diseases. The immunoglobulins disclosed herein can be of any type (e.g., IgG, IgE, IgM, IgD, and IgA), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass of immunoglobulin molecule. Immunoglobulins can be derived from any species. However, in one aspect the immunoglobulin is of human, murine or rabbit origin.

[0023] As used herein, the term "antibody fragment(s)" includes a portion of a full-length antibody, generally the antigen-binding or variable region thereof. Exemplary antibody fragments include Fab, Fab', F(ab') 2 , and Fv fragments; diabodies; linear antibodies; minibodies (Olafsen et al. (2004) "Protein Eng. Design & Sel" 17(4):315-323), fragments produced by Fab expression libraries, anti-idiotypic (anti-Id) antibodies, CDRs (complementarity determining regions), and epitope-binding fragments of any of the above, single-chain antibody molecules that immunospecifically bind to cancer cell, viral, or microbial antigens; Multispecific antibodies are included.

[0024] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies that make up the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to polyclonal antibody preparations, which include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, monoclonal antibodies are advantageous in that they can be synthesized without contamination with other antibodies. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies used in accordance with the subject matter described herein may be made by the hybridoma method first described by Kohler et al. (1975) Nature, 256:495, or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567; U.S. Pat. No. 5,807,715). "Monoclonal antibodies" can also be isolated from phage antibody libraries, for example, using the techniques described in Clackson et al. (1991) Nature 352:624-628; Marks et al. (1991) J. Mol. Biol. 222:581-597.

[0025] Monoclonal antibodies herein specifically include "chimeric" antibodies, and fragments of such antibodies, in which a portion of the heavy and / or light chain is identical or homologous to corresponding sequences in an antibody from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical or homologous to corresponding sequences in an antibody from another species or belonging to another antibody class or subclass, which exhibit the desired biological activity. (US Pat. No. 4,816,567; and Morrison et al. (1984) Proc. Natl. Acad. Sci. USA, 81:6851-6855). Chimeric antibodies of interest herein include "primatized" antibodies comprising variable domain antigen-binding sequences derived from a non-human primate (e.g., Old World Monkey, Ape, etc.) and human constant region sequences.

[0026] The term "chimeric" antibody refers to antibodies in which a portion of the heavy and / or light chain is derived from a particular source or species and the remainder of the heavy and / or light chain is derived from a different source or species.

[0027] The "class" of an antibody refers to the type of constant domain or region possessed by its heavy chains. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, some of which are subclasses (isotypes), such as IgG 1 , IgG 2 , IgG 3 , IgG 4 , IgA 1 , and IgA 2 can be further divided into The heavy-chain constant domains that correspond to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[0028] The term "intact antibody" as used herein comprises the VL and VH domains as well as the light chain constant domain (CL) and heavy chain constant domains CH1, CH2 and CH3. The constant domains may be native sequence constant domains (eg, human native sequence constant domains) or amino acid sequence variant thereof. An intact antibody may possess one or more "effector functions," which refer to biological activities attributable to the Fc constant region (a native sequence Fc region or amino acid sequence variant Fc region) of an antibody. Examples of antibody effector functions include C1q binding, complement-dependent cytotoxicity, Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, and downregulation of cell surface receptors such as B cell receptors and BCR.

[0029] As used herein, the term "Fc region" is used to define a C-terminal region of an immunoglobulin heavy chain that contains at least a portion of the constant region. The term includes native sequence Fc regions and variable Fc regions. In one embodiment, the human IgG heavy chain Fc region extends from Cys226 or from Pro230 to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) of the Fc region may or may not be present. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region is according to the EU numbering system (also called EU index), as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991.

[0030] The term "framework" or "FR" as used herein refers to variable domain residues other than hypervariable region (HVR) residues. The FRs of variable domains generally consist of four FR domains: FR1, FR2, FR3 and FR4. Thus, HVR and FR sequences generally appear in the next sequence in VH (or VL). FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

[0031] The terms "full-length antibody," "intact antibody," and "whole antibody" are used interchangeably herein and refer to an antibody having a heavy chain that has a structure substantially similar to that of a native antibody or that contains an Fc region as defined herein.

[0032] A "human antibody" is one that possesses amino acid sequences that correspond to the amino acid sequences of an antibody, or other human antibody coding sequences, produced by humans or human cells, or derived from non-human sources utilizing human antibody repertoires. This definition of human antibody specifically excludes humanized antibodies that contain non-human antigen-binding residues.

[0033] A "humanized" antibody refers to a chimeric antibody comprising amino acid residues from non-human HVRs and amino acid residues from human FRs. In certain embodiments, a humanized antibody comprises substantially all of at least one, typically two, variable domains, in which all or substantially all of the HVRs (e.g., CDRs) correspond to those of a non-human antibody and all or substantially all of the FRs correspond to those of a human antibody. A humanized antibody may optionally comprise at least a portion of an antibody constant region derived from a human antibody. A "humanized form" of an antibody refers to an antibody that has been humanized, eg, a non-human antibody.

[0034] An "isolated antibody" is an antibody that has been separated from a component of its natural environment. In some embodiments, the antibody is purified to greater than 95% or greater than 99% purity, e.g., as determined by electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reverse phase HPLC). For a review of methods for assessment of antibody purity, see, eg, Flatman et al., J. Chromatogr. B, 848:79-87 (2007).

[0035] "Isolated nucleic acid" refers to a nucleic acid molecule that is separated from a component of its natural environment. An isolated nucleic acid includes a nucleic acid molecule in which the nucleic acid molecule is present extrachromosomally or in a chromosomal location different from its natural chromosomal location, although originally contained in cells containing the nucleic acid molecule.

[0036] An "isolated nucleic acid encoding an antibody" refers to one or more nucleic acid molecules encoding antibody heavy and light chains (or fragments thereof), including the nucleic acid molecule(s) in a single vector or separate vectors, such nucleic acid molecule(s) being present in one or more locations within a host cell.

[0037] A "naked antibody" refers to an antibody that is not conjugated to a heterologous moiety (eg, a cytotoxic moiety) or radiolabel. A naked antibody may be present in a pharmaceutical formulation.

[0038] "Native antibody" refers to naturally occurring immunoglobulin molecules with varying structures. For example, native IgG antibodies are heterotetrameric glycoproteins of approximately 150,000 daltons, composed of two identical light chains and two identical heavy chains that are disulfide-bonded. From N-terminus to C-terminus, each heavy chain has a variable region (VH), also called a variable heavy domain or heavy chain variable domain, followed by three constant domains (CH1, CH2, and CH3). Similarly, from N-terminus to C-terminus, each light chain has a variable region (VL), also called a variable light domain or light chain variable domain, followed by a constant light (CL) domain. The light chains of antibodies can be assigned to one of two types, called kappa (κ) and lambda (λ), based on the amino acid sequences of their constant domains.

[0039] A "percent (%) amino acid sequence identity" with respect to a reference polypeptide sequence is a measure of sequence identity after aligning the sequences and introducing gaps, if necessary, to obtain the maximum percent sequence identity. It is defined as the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in the reference polypeptide sequence, not considered a fraction. Alignments for purposes of determining percent amino acid sequence identity may be performed by a variety of methods within the skill in the art, such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. can be accomplished using computer software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve maximal degree of alignment over the entire length of the sequences being compared. However, for purposes herein, % amino acid sequence identity values ​​are generated using the sequence comparison computer program ALIGN-2. The ALIGN-2 sequence comparison computer program was created by Genentech, Inc. and the source code is filed in the User Documentation of the U.S. Copyright Office, Washington D.C., 20559, under U.S. Copyright Registration No. TXU510087. Registered. The ALIGN-2 program is publicly available from Genentech, Inc. (South San Francisco, Calif.) or may be compiled from its source code. ALIGN-2 programs should be compiled for use on UNIX operating systems, including Digital UNIX V4.0D. All sequence comparison parameters were set by the ALIGN-2 program and remained unchanged.

[0040] In situations where ALIGN-2 is employed for amino acid sequence comparison (or has or contains a certain % amino acid sequence identity to, with, or against a given amino acid sequence B) The % amino acid sequence identity of a given amino acid sequence A (which may be purchased as a given amino acid sequence A) to a given amino acid sequence B, with or against sequence B is: Calculated to: 100 times the fraction X / Y where X is the number of amino acid residues scored as identity matches by the sequence alignment program ALIGN-2 in this program's alignment of A and B, and Y is the total number of amino acid residues in B is. It will be understood that the % amino acid sequence identity of A to B is not equal to the % amino acid sequence identity of B to A if the length of amino acid sequence A is not equal to the length of amino acid sequence B. Unless otherwise stated, all % amino acid sequence identity values ​​used herein are obtained as described in the immediately preceding paragraph using the ALIGN-2 computer program.

[0041] Depending on the amino acid sequence of the constant domain of their heavy chains, intact antibodies can be assigned to different "classes." There are five major classes of intact immunoglobulin antibodies, namely IgA, IgD, IgE, IgG, and IgM, some of which have "subclasses" (isotypes) such as IgG1, IgG2, It can be further divided into IgG3, IgG4, IgA1 and IgA2. The heavy-chain constant domains that correspond to the different classes of antibodies are called α, δ, ε, γ, and μ, respectively. The subunit structures and three-dimensional arrangements of different classes of immunoglobulins are well known. Ig forms include hinge-modified or non-hinge forms (Roux et al. (1998) J. Immunol. 161:4083-4090; Lund et al. (2000) Eur. J. Biochem. 267: 7246-7256; US Patent Application Publication No. 2005 / 0048572; US Patent Application Publication No. 2004 / 0229310).

[0042] As used herein, the term "human consensus framework" refers to a framework that represents the most commonly occurring amino acid residues in a selection of human immunoglobulin VL or VH framework sequences. Generally, the selection of human immunoglobulin VL or VH sequences is from a subgroup of variable domain sequences. In general, subgroups of sequences are as in Kabat et al., "Sequences of Proteins of Immunological Interest," 5th ed., NIH Publication, pp. 91-3242, Bethesda, Md. (1991), vols. subgroup. In one embodiment, for VL, the subgroup is subgroup kappa I in Kabat et al. (see above). In one embodiment, for VH, the subgroup is subgroup III in Kabat et al. (see above).

[0043] An "acceptor human framework" for the purposes of this specification is a light chain variable domain (VL) framework or heavy chain variable domain (VL) framework derived from a human immunoglobulin framework or a human consensus framework, defined below. A framework containing the amino acid sequence of a domain (VH) framework. An acceptor human framework "derived from" a human immunoglobulin framework or a human consensus framework may contain that same amino acid sequence or may contain changes in the amino acid sequence. In some embodiments, the number of amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less. In some embodiments, the VL acceptor human framework is identical in sequence to a VL human immunoglobulin framework sequence or a human consensus framework sequence.

[0044] The terms "variable region" or "variable domain" as used herein refer to the domains of antibody heavy or light chains involved in the binding of the antibody to antigen. The heavy and light chain variable domains (VH and VL, respectively) of naturally occurring antibodies have a generally similar structure, with each domain consisting of four conserved framework regions (FR) and three and hypervariable regions (HVR). See, for example, Kindt et al., Kuby Immunology, 6 th , W.H. Freeman and Co., page 91 (2007). A single VH or VL domain may be sufficient to confer antigen binding specificity. Additionally, antibodies that bind a particular antigen may be isolated using the VH or VL domain of the antibody that binds the antigen and screening a library of complementary VL or VH domains, respectively. See, eg, Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[0045] The term "hypervariable region" or "HVR", as used herein, is hypervariable in sequence and / or forms structurally defined loops ("hypervariable loops"), Refers to each of the regions of an antibody variable domain. Generally, a native four-chain antibody contains 6 HVRs, 3 in VH (H1, H2, H3) and 3 in VL (L1, L2, L3). HVRs generally contain amino acid residues from hypervariable loops and / or from "complementarity determining regions" (CDRs), which have the highest sequence variability and / or are involved in antigen recognition. are doing. Exemplary hypervariable loops are amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2) and 96-101 ( H3). (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987).) Exemplary CDRs (CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3) is located at amino acid residues 24-34 of L1, 50-56 of L2, 89-97 of L3, 31-35B of H1, 50-65 of H2 and 95-102 of H3. (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)) Except for CDR1 in VH, CDRs are generally amino acids that form hypervariable loops. Contains residues. CDRs also contain "specificity determining regions," or "SDRs," which are residues that contact antigen. The SDR is contained within a region of the CDR called the abbreviated-CDR or a-CDR. Exemplary a-CDRs (a-CDR-L1, a-CDR-L2, a-CDR-L3, a-CDR-H1, a-CDR-H2 and a-CDR-H3) are 31-34 of L1 , 50-55 of L2, 89-96 of L3, 31-35B of H1, 50-58 of H2 and 95-102 of H3. (See Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008).) Unless otherwise specified, HVR residues, and other residues in the variable domain (e.g., FR residues), are as described in Kabat et al. numbered herein according to (supra).

[0046] "Effector functions" refer to those biological activities attributable to the Fc region of an antibody that vary according to the isotype of the antibody. Examples of antibody effector functions include the following. Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (eg, B-cell receptors); and B-cell activation.

[0047] The term "epitope" refers to the specific site on the antigen molecule to which an antibody binds.

[0048] "Epitope 4D5" or "4D5 epitope" or "4D5" is the region within the extracellular domain of HER2 to which antibody 4D5 (ATCC CRL 10463) and trastuzumab bind. This epitope is close to the transmembrane domain of HER2 and within domain IV of HER2. To screen for antibodies that bind to the 4D5 epitope, routine cross-blocking assays such as those described in "Antibodies, A Laboratory Manual," Cold Spring Harbor Laboratory, Harlow and David Lane, eds. (1988) can be performed. Alternatively, epitope mapping can be performed to assess whether the antibody binds to the 4D5 epitope of HER2 (any one or more residues within a region from about residue 550 to about residue 610 that includes HER2 (SEQ ID NO:39)).

[0049] "Epitope 2C4" or "2C4 epitope" is the region within the extracellular domain of HER2 to which antibody 2C4 binds. To screen for antibodies that bind to the 2C4 epitope, routine cross-blocking assays such as those described in "Antibodies, A Laboratory Manual", Cold Spring Harbor Laboratory, Harlow and David Lane, eds. (1988) can be performed. Alternatively, epitope mapping can be performed to assess whether the antibody binds to the 2C4 epitope of HER2. Epitope 2C4 comprises residues from domain II in the extracellular domain of HER2. The 2C4 antibody and pertuzumab bind to the extracellular domain of HER2 at the junction of domains I, II and III (Franklin et al. Cancer Cell 5:317-328 (2004)).

[0050] "Affinity" refers to the strength of the total non-covalent interactions between a single binding site on a molecule (eg antibody) and its binding partner (eg antigen). Unless otherwise stated, "binding affinity" as used herein refers to specific binding affinity reflecting a 1:1 interaction between members of a binding pair (eg, antibody and antigen). The affinity of molecule X for its partner Y can generally be expressed by the dissociation constant (Kd). Affinity can be measured by methods common in the art, including those described herein. Specific illustrative descriptions and exemplary embodiments for measuring binding affinity are described below. In certain embodiments, the antibodies described herein are <1 μM, <100 nM, <10 nM, <5 nm, <4 nM, <3 nM, <2 nM, <1 nM, <0.1 nM, <0.01 nM, or <0.001 nM (e.g., 10 -8 M or less, e.g. 10 -8 M~10 -13 M, for example 10 -9 M~10 -13 M) dissociation constant (Kd).

[0051] An "affinity matured" antibody refers to an antibody that has one or more alterations in one or more hypervariable regions (HVRs) relative to a parent antibody that does not have such alterations, such alterations improving the affinity of the antibody for its antigen.

[0052] The term "vector," as used herein, refers to a nucleic acid molecule capable of propagating another nucleic acid to which it has been linked. The term includes vectors as self-replicating nucleic acid structures and vectors that have integrated into the genome of a host cell into which they are introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operably linked. Such vectors are referred to herein as "expression vectors".

[0053] As used herein, the term "free cysteine ​​amino acid" refers to a cysteine ​​amino acid residue that has been engineered into the parent antibody, has a thiol functionality (-SH), and is unpaired as an intramolecular or intermolecular disulfide bridge. The term "amino acid" as used herein means glycine, alanine, valine, leucine, isoleucine, phenylalanine, proline, serine, threonine, tyrosine, cysteine, methionine, lysine, arginine, histidine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine or citrulline.

[0054] The terms "linker," "linker unit," or "linkage," as used herein, refer to a chemical moiety comprising a chain of atoms that covalently bonds a CIDE moiety to an antibody or a chain of atoms that covalently bonds a component of CIDE to another component of CIDE. In various embodiments, the linker is a divalent group identified as L1 or L2.

[0055] A "patient" or "individual" or "subject" is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cattle, sheep, cats, dogs, horses, etc.), primates (e.g., non-human primates such as humans and monkeys), rabbits, rodents (e.g., mice, rats, etc.). In certain embodiments, the patient, individual, or subject is human. In some embodiments, the patient may be a "cancer patient," ie, one having or at risk of having one or more symptoms of cancer.

[0056] A "patient population" refers to a group of cancer patients. Populations such as these can be used to demonstrate statistically significant efficacy and / or safety of drugs.

[0057] A "relapsed" patient is one who has signs or symptoms of cancer after remission. Patients relapsed after adjuvant or neoadjuvant therapy, as appropriate.

[0058] A cancer or biological sample that "exhibits HER expression, amplification, or activation" is one that expresses (including overexpression) the HER receptor, contains an amplified HER gene, and / or otherwise demonstrates HER receptor activation or phosphorylation in a diagnostic test.

[0059] As used herein, "neoadjuvant therapy" or "preoperative therapy" refers to therapy given prior to surgery. The goal of neoadjuvant therapy is to provide immediate systemic treatment, potentially eradicating growing micrometastases when a standard course of surgery followed by systemic therapy is followed. Neoadjuvant therapy can also help reduce tumor size, thereby allowing complete resection of initially unresectable tumors or preservation of parts of organs and their functions. Furthermore, neoadjuvant therapy allows in vivo assessment of drug efficacy that can guide subsequent treatment selection.

[0060] As used herein, "adjuvant therapy" refers to therapy given after definitive surgery in which no evidence of residual disease can be detected in order to reduce the risk of disease recurrence. The purpose of adjuvant therapy is to prevent cancer recurrence and thus reduce the likelihood of cancer-related death. Adjuvant therapy herein specifically excludes neoadjuvant therapy.

[0061] "Definitive surgery" is used as the term is used within the medical community. Definitive surgery includes, for example, surgery, surgical or other procedures that result in removal or resection of a tumor, including those that result in removal or resection of all grossly visible tumors. Radical surgery includes, for example, complete or curative resection or total gross resection of the tumor. Definitive surgery includes procedures that occur in one or more stages, including multi-stage surgical procedures in which one or more surgical or other procedures are performed prior to resection of the tumor. Definitive surgery includes procedures to remove or resect the tumor, including the organs, parts of organs and tissues involved, and surrounding organs such as lymph nodes, parts of organs or tissues. Removal may be incomplete such that tumor cells may remain undetected.

[0062] "Survival" refers to patients remaining alive and includes disease-free survival (DFS), progression-free survival (PFS) and overall survival (OS). Survival can be estimated by the Kaplan-Meier method and survival differences are calculated using the stratified log-rank test.

[0063] "Progression-free survival" (PFS) is the time from day 1 of treatment to documented disease progression (including isolated CNS progression) or death from any cause under study, whichever occurs first.

[0064] "Disease-free survival (DFS)" refers to patients who have survived without cancer recurrence for a defined period of time, such as about 1 year, about 2 years, about 3 years, about 4 years, about 5 years, about 10 years, from the start of treatment or initial diagnosis. In one aspect of the subject matter described herein, DFS is analyzed according to intent-to-treat principles, ie, patients are evaluated based on assigned therapy. Events used in the analysis of DFS can include local, regional and distant recurrence of cancer, development of second cancers, and death from any cause in patients without prior events (e.g., breast cancer recurrence or second primary cancer).

[0065] "Overall survival" refers to patients living for a defined period of time, such as about 1, about 2, about 3, about 4, about 5, about 10 years from initiation of treatment or initial diagnosis.

[0066] By "prolonging survival" is meant increasing DFS and / or OS in treated patients compared to untreated patients or compared to a control treatment protocol. Survival is monitored after initiation of treatment or after initial diagnosis, such as for at least about 6 months, or at least about 1 year, or at least about 2 years, or at least about 3 years, or at least about 4 years, or at least about 5 years, or at least about 10 years.

[0067] "Monotherapy" means a therapeutic regimen that includes only a single therapeutic agent for the treatment of cancer or tumor during a course of treatment.

[0068] "Maintenance therapy" means a therapeutic regimen given to reduce the likelihood of disease recurrence or progression. Maintenance therapy can be provided for any length of time, including extended periods for the life of the subject. Maintenance therapy may be provided after initial therapy or in combination with initial therapy or additional therapy. Dosages used for maintenance therapy may vary, and may include reduced dosages compared to dosages used for other types of therapy.

[0069] The terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to a cell into which exogenous nucleic acid has been introduced, including progeny of such cells. A host cell includes "transformants" and "transformed cells," which include the primary transformed cell and progeny derived therefrom regardless of passage number. Progeny may not be completely identical in nucleic acid content to the parent cell, but may contain mutations. Mutant progeny that have the same function or biological activity as screened for or selected for the originally transformed cell are included in the present invention.

[0070] The terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth / proliferation. A "tumor" includes one or more cancer cells. Examples of cancer are provided elsewhere herein.

[0071] A "HER2-positive" cancer includes cancer cells that have higher than normal levels of HER2. Examples of HER2-positive cancers include HER2-positive breast cancer and HER2-positive gastric cancer. Optionally, the HER2-positive cancer has an immunohistochemistry (IHC) score of 2+ or 3+ and / or an in situ hybridization (ISH) amplification rate of ≧2.0. The term "HER2-positive cell" refers to a cell that expresses HER2 on its surface.

[0072] The terms "early stage breast cancer (EBC)" or "early stage breast cancer" are used herein to refer to breast cancer that has not spread beyond the breast or axillary lymph nodes. This includes ductal carcinoma in situ and stages I, IIA, IIB and IIIA breast cancer.

[0073] References to tumors or cancers as "Stage 0," "Stage I," "Stage II," "Stage III," or "Stage IV," and to various substages within this classification, refer to the classification of tumors or cancers using full stage classification or Roman numeral staging systems known in the art. The actual stage of cancer depends on the type of cancer, but in general, stage 0 cancer is an in situ lesion, stage I cancer is a small localized tumor, stage II and III cancer is locally advanced tumor showing local lymph node involvement, and stage IV cancer represents metastatic cancer. The specific stages of each type of tumor are known to those skilled in the art.

[0074] The term "metastatic breast cancer" refers to the condition of breast cancer in which cancer cells travel from their original site to one or more sites elsewhere in the body by blood or lymphatic vessels to form one or more secondary tumors in one or more organs other than the breast.

[0075] "Advanced" cancer is cancer that has spread outside the site or organ of origin, either by local invasion or by metastasis. Thus, the term "advanced" cancer includes both locally advanced and metastatic disease. A "recurrent" cancer is one that has regrowth, either at the initial site or at a distant site, after response to initial therapy such as surgery. A "locally recurrent" cancer is a cancer that recurs after treatment in the same place as a previously treated cancer. A "operable" or "resectable" cancer is a cancer confined to the primary organ and amenable to surgery (resection). A "non-resectable" or "unresectable" cancer cannot be removed (excised) by surgery.

[0076] As used herein, the term "cytotoxic agent" refers to a substance that inhibits or prevents cell function and / or causes cell death or destruction. Cytotoxic agents include radioactive isotopes (e.g., At 211 , I 131 , I 125 , Y 90 , Re 186 , Re 188 , Sm 153 , Bi 212 , P 32 , Pb 212 chemotherapeutic agents or drugs (e.g. methotrexate, adriamycin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents); growth inhibitors; enzymes and fragments thereof, e.g. fragments and / or variants thereof); and the various anti-tumor or anti-cancer agents disclosed below.

[0077] A "chemotherapeutic agent" refers to a chemical substance useful in the treatment of cancer. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (CYTOXAN®); alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodo-pa, carboquone, methledopa ( meturedopa), and uredo-pa; ethyleneimines and methylamelamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylomelamines; acetogenins (especially bratacin and bratacinone); delta-9-tetrahydrocannabinol (dronabinol, MARINOL®); beta-lapachone; lapachol; colchicine; betulinic acid; , CAMPTOSAR®), acetylcamptothecin, scopolectin, and 9-aminocamptothecin); bryostatin; callistatin; CC-1065 (including its adzelesin, carzelesin and vizelesin synthetic analogues); podophyllotoxin; teniposide; cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycins (including synthetic analogues, KW-2189 and CB1-TM1); erythrobin; mustards such as chlorambucil, chlornafadine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novenvitine, phenesterin, prednimustine, trophosphamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine and ranimnustine; antibiotics such as engine antibiotics (e.g. calicheamicins, especially calicheamicin γ1I and calicheamicin omega I1 (e.g. Nicolaou et al., Angew.Chem Intl.Ed.Engl., 33) : 183-186 (1994)); CDP323, an oral alpha-4 integrin inhibitor; dynemicins, including dynemicin A; esperamycin; chromophore), aclacinomycin, actinomycin, anthramycin, azaserine, bleomycin, cactinomycin, carabicin, caminomycin, cardinophylline, chromomycin, dactinomycin, daunorubicin, detrubicin, 6-diazo-5-oxo-L - norleucine, doxorubicin (ADRIAMYCIN®, morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, doxorubicin HCl liposome injection (DOXIL®), liposomal doxorubicin TLC D-99 (MYOCET®) )), pegylated liposomal doxorubicin (CAELYX®), and deoxydoxorubicin), epirubicin, ethorubicin, idarubicin, marceromycin, mitomycins such as mitomycin C, mycophenolic acid, nogaramycin, olibomycin, peplomycin, porphyromycin , puromycin, keramycin, rhodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, dinostatin, zorubicin; antimetabolites such as methotrexate, gemcitabine (GEMZAR®), tegafur (UFTORAL®), capecitabine ( XELODA®), epothilones, and 5-fluorouracil (5-FU); folate analogs such as denopterin, methotrexate, pteropterin, trimetrexate; pridine analogs such as fludarabine, 6-mercaptopurine, thiamipurine, thioguanine; pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxfluridine, enocitabine, floxuridine; androgens such as carsterone, drostanolone propionate, epithiostanol, mepitiostane, testolactone; anti-adrenal agents such as aminoglutethimide, mitotane, trilostane; folic acid supplements such as folinic acid; acegratone; aldophosphamide glycosides; aminolevulinic acid; diaziquone; elfornithine; elliptinium acetate; epothilone; etogluside; gallium nitrate; hydroxyurea; lentinan; Pirarubicin; Losoxantrone; 2-Ethylhydrazide; Procarbazine; PSK® Polysaccharide Complex (JHS Natural Products, Eugene, Oreg.); Lazoxane; ',2'-trichlorotriethylamine; trichothecenes (particularly T-2 toxin, veracrine A, roridin A and anguidine); urethanes; vindesines (ELDISINE®, FILDESIN®); dacarbazine; thiotepa; taxoids such as paclitaxel (TAXOL®), albumin-modified nanoparticle formulations of paclitaxel (ABRAXANETM), and docetaxel (TAXOTERE®); chlorambucil 6-thioguanine; mercaptopurine; methotrexate; platinum agents such as cisplatin, oxaliplatin (e.g., ELOXATIN®), and carboplatin; vinblastine (VELBAN®), vincristine (ONCOVIN®), vindesine Vincas that prevent tubulin polymerization and microtubule formation, including (ELDISINE®, FILDESIN®), and vinorelbine (NAVELBINE®); etoposide (VP-16); ifosfamide mitoxantrone; leucovorin; novantrone; edatrexate; daunomycin; aminopterin; ibandronate; retinoic acid; bisphosphonates such as clodronate (e.g. BONEFOS® or OSTAC®), etidronate (DIDROCAL®), NE-58095, zoledronic acid / zoledronate (ZOMETA®), alendronate (FOSAMAX®), pamidronate (AREDIA®), tiludronate (SKELID®), or risedronate (ACTONEL®); troxacitabine (1,3-dioxolane nucleoside cytosine analog); sense oligonucleotides, particularly genes in signaling pathways involved in abnormal cell proliferation, such as PKC-alpha, Raf, H-Ras, and epidermal growth factor receptor (EGF-R); vaccines, such as THERATOPE® Vaccines and gene therapy vaccines, such as ALLOVECTIN®, LEUVECTIN®, and VAXID® vaccines; topoisomerase 1 inhibitors (e.g., LURTOTECAN®); rmRH (e.g., ABARELIX ( BAY439006 (sorafenib, Bayer); SU-11248 (sunitinib, SUTENT®, Pfizer); perifosine, COX-2 inhibitors (e.g., celecoxib or etoricoxib), proteosome inhibitors (e.g., PS341); Bortezomib (VELCADE®); CCI-779; Tipifarnibi (R11577); Olafenib, ABT510; EGFR inhibitors (see definitions below); tyrosine kinase inhibitors; serine-threonine kinase inhibitors such as rapamycin (sirolimus, RAPAMUNE®); farnesyl transferase inhibitors such as lonafarnib (SCH 6636, SARASARTM) and pharmaceutically acceptable salts, acids or derivatives of any of the above; and combinations of two or more of the above, such as CHOP ((abbreviation for combination therapy of cyclophosphamide, doxorubicin, vincristine, prednisolone), FOLFOX (Oxaliplatin (ELOXATINTM) and 5-FU, an abbreviation for combination therapy with leucovorin).

[0078] Chemotherapeutic agents, as defined herein, include "anti-hormonal agents" or "endocrine therapeutic agents" that act to modulate, reduce, block, or inhibit the effects of hormones that may promote cancer growth. They may themselves be hormones and antiestrogens with mixed agonist / antagonist profiles, including selective estrogen receptor modulators (SERMs) such as tamoxifen (NOLVADEX®), 4-hydroxy tamoxifen, toremifene (FARESTON®), idoxifene, droloxifene, raloxifene (EVISTA®), trioxyphene, keoxifene, and SERM3; fulvestrant; (FASLODEX®), and EM800 (such agents may block estrogen receptor (ER) dimerization, inhibit DNA binding, increase ER turnover, and / or suppress ER levels); steroidal aromatase inhibitors, such as formestane and exemestane (AROMASIN®), and anastrazole (ARIMIDEX®), letrozole ( FEMARA®), and nonsteroidal aromatase inhibitors such as aminoglutethimide, and other aromatase inhibitors, including vorozole (RIVISOR®), megestrol acetate (MEGASE®), fadrozole, and 4(5)-imidazole; leuprolide (LUPRON® and ELIGARD®), goserelin, buserelin, and tripterelin. progestins such as megestrol acetate and medroxyprogesterone acetate; estrogens such as diethylstilbestrol and premarin; sex steroids, including androgens / retinoids such as fluoxymesterone, all transretionic acids, and fenretinide; onapristone; D); antiandrogens such as flutamide, nilutamide, and bicalutamide; and pharmaceutically acceptable salts, acids, or derivatives of any of the above; and combinations of two or more of the above.

[0079] The term "immunosuppressant," as used herein with respect to adjuvant therapy, refers to substances that act to suppress or mask the immune system of the mammal being treated herein. This would include substances that suppress cytokine production, down-regulate or suppress self-antigen expression, or mask MHC antigens. Examples of such agents include 2-amino-6-aryl-5-substituted pyrimidines (see U.S. Pat. No. 4,665,077); nonsteroidal anti-inflammatory drugs (NSAIDs); glucocorticoids such as ganciclovir, tacrolimus, cortisol or aldosterone; purine antagonists such as cophenolate mofetil (MMF); alkylating agents such as cyclophosphamide; bromocriptine; danazol; dapsone; corticoid analogs, such as prednisone, methylprednisolone, such as SOLU-MEDROL® methylprednisolone sodium succinate and dexamethasone; dihydrofolate reductase inhibitors, such as methotrexate (oral or subcutaneous); antimalarial agents such as chloroquine and hydroxy; sulfasalazine; Cytokine or cytokine receptor antibodies, including infliximab (REMICADE®) or adalimumab, anti-TNF-alpha immune adesin (etanercept), anti-TNF-β antibodies, anti-interleukin-2 (IL-2) antibodies and anti-IL-2 receptor antibodies, and anti-interleukin-6 (IL-6) receptor antibodies and antagonists (such as ACTEMRA™ tocilizumab); anti-LF, including anti-CD11a and anti-CD18 antibodies; A-1 antibody; anti-L3T4 antibody; heterologous anti-lymphocyte globulin; pan-T antibody, preferably anti-CD3 or anti-CD4 / CD4a antibody; deoxyspergualin; rapamycin; T-cell receptor (Cohen et al., U.S. Patent No. 5,114,721); T-cell receptor fragment (Offner et al., Science 251:430-432 (1991); WO90 / 11294; Ianeway, Nature, 341:482 (1989); ); BAFF antagonists, such as BAFF and BR3 antibodies and zTNF4 antagonists (for review, see Mackay and Mackay, Trends Immunol, 23:113-5 (2002); see also definitions below); , Durie et al. Science 261:1328-30 (1993); Mohan et al. J. Immunol 154:1470-80 (1995)) and CTLA4-Ig (Finck et al. Science 265:1225-7 (1994)); 10B9 can be mentioned. Some preferred immunosuppressants herein include cyclophosphamide, chlorambucil, azathioprine, leflunomide, MMF, or methotrexate.

[0080] As used herein, “treatment” (and grammatical variants thereof such as “treating” or “treating”) refers to clinical intervention in an attempt to alter the course of treatment in the individual being treated, and can be for prophylaxis or during the course of clinical pathology. Desirable effects of treatment include preventing the onset or recurrence of disease, alleviating symptoms, attenuating any direct or indirect pathological consequences of disease, preventing metastasis, reducing the rate of disease progression, remission or alleviation of disease, and remission or improved prognosis. In some embodiments, the subject antibodies even described herein are used to delay the onset of disease or slow progression of disease.

[0081] A drug that is administered "concurrently" with one or more other drugs is administered during the same treatment cycle, on the same treatment days as the one or more other drugs, and optionally at the same time as the one or more other drugs. For example, for cancer therapy given every 3 weeks, each co-administered drug is administered on day 1 of the 3-week cycle.

[0082] An "effective amount" of an agent, eg, pharmaceutical formulation, refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result. For example, an effective amount of a drug for treating cancer may reduce the number of cancer cells, reduce tumor size, inhibit (i.e., slow, preferably stop) cancer cell invasion into peripheral organs, inhibit (i.e., slow, preferably stop) tumor metastasis, inhibit (i.e., slow, preferably stop), inhibit tumor growth, and / or alleviate one or more of the symptoms associated with cancer. To the extent the drug may prevent the growth of and / or kill existing cancer cells, the drug may be cytostatic and / or cytotoxic. Effective amounts increase progression-free survival (e.g., as measured by Response Evaluation Criteria in Solid Tumors (RECIST) or CA-125 changes), produce objective responses (partial response (PR) or complete response (CR)), prolong overall survival, and / or ameliorate one or more symptoms of cancer (e.g., as assessed by FOSI).

[0083] As used herein, the term "therapeutically effective amount" means any amount that results in the treatment of a disease, disorder, or side effect, or slows the rate of progression of the disease or disorder, as compared to a corresponding subject not receiving such amount. The term also includes within its scope amounts effective to enhance normal physiological function. For therapeutic use, therapeutically effective amounts of Ab-CIDE and salts thereof can be administered as the raw chemical. Additionally, the active ingredient may be presented as a pharmaceutical composition.

[0084] As used herein, unless otherwise defined in the claims, the term "optionally" means that the subsequently described event(s) may or may not occur, and includes both the event(s) that do and the event(s) that do not occur.

[0085] As used herein, unless otherwise defined, the phrases "optionally substituted," "substituted," or variations thereof refer to optional substitution with one or more substituents, including multiple degrees of substitution, such as 1, 2 or 3. This phrase should not be construed as redundant with the substitutions described and illustrated herein.

[0086] The term "pharmaceutical formulation" refers to a preparation that is in a form such that the biological activity of the active ingredients contained in the preparation is effective and that does not contain additional components that are unacceptably toxic to the subject to whom the formulation is administered.

[0087] "Pharmaceutically acceptable excipient" refers to an ingredient in a pharmaceutical formulation other than the active ingredient that is non-toxic to the subject. Pharmaceutically acceptable excipients include, but are not limited to, buffers, carriers, stabilizers, or preservatives.

[0088] As used herein, the phrase "pharmaceutically acceptable salt" means pharmaceutically acceptable organic or inorganic salts of a molecule. Representative salts include sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, saccharate, formate. acid salts, benzoates, glutamates, methanesulfonates, ethanesulfonates, benzenesulfonates, p-toluenesulfonates, and pamoates (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)). A pharmaceutically acceptable salt may include another molecule such as an acetate, succinate or other counterion. A counterion can be any organic or inorganic moiety that stabilizes the charge of the parent compound. Additionally, a pharmaceutically acceptable salt may have more than one charged atom in its structure. Instances where multiple charged atoms are part of the pharmaceutically acceptable salt can have multiple counter ions. Accordingly, a pharmaceutically acceptable salt can have one or more charged atoms and / or one or more counterions.

[0089] Other non-pharmaceutically acceptable salts may be useful in the preparation of the compounds described herein and these should be considered forming a further aspect of the subject matter. These salts, such as oxalates or trifluoroacetates, are not themselves pharmaceutically acceptable, but can be useful in the preparation of salts useful as intermediates in obtaining the compounds described herein and their pharmaceutically acceptable salts.

[0090] As used herein, the term "plurality" refers to two or more conjugates. Each conjugate may be the same or different from any other conjugate in the plurality.

[0091] "Small molecule" or "small molecule compound" generally refers to organic molecules less than about 5 kilodaltons (Kd) in size. In some embodiments, small molecules are less than about 4Kd, 3Kd, about 2Kd, or about 1Kd. In some embodiments, small molecules are less than about 800 Daltons (D), about 600D, about 500D, about 400D, about 300D, about 200D, or about 100D. In some embodiments, small molecules are less than about 2000 g / mol, less than about 1500 g / mol, less than about 1000 g / mol, less than about 800 g / mol, or less than about 500 g / mol. In some embodiments, small molecules are non-polymeric. Small molecules are not proteins, polypeptides, oligopeptides, peptides, polynucleotides, oligonucleotides, polysaccharides, glycoproteins, proteoglycans, and the like. Small molecule derivatives refer to molecules that share the same structural core as the original small molecule, but can be prepared by a series of chemical reactions from the original small molecule.

[0092] As used herein, the term "alkyl" is anywhere from 1 to 12 carbon atoms in length (C 1 ~C 12 ), wherein the alkyl radical is optionally substituted independently with one or more substituents described below. In another embodiment, the alkyl radical has 1-8 carbon atoms (C 1 ~C 8 ), or 1 to 6 carbon atoms (C 1 ~C 6 ). Examples of alkyl groups include, but are not limited to, methyl l(Me, -CH 3 ), ethyl (Et-CH 2 CH 3 ), 1-propyl (n-PR, n-propyl, -CH 2 CH 2 CH 3 ), 2-propyl (i-PR, i-propyl, -CH(CH 3 ) 2 ), 1-butyl (n-Bu, n-butyl, -CH 2 CH 2 CH 2 CH 3 ), 2-methyl-1-propyl (i-Bu, i-butyl, -CH 2 CH(CH 3 ) 2 ), 2-butyl (s-Bu, s-butyl, -CH(CH 3 )CH 2 CH 3 ), 2-methyl-2-propyl (t-Bu, t-butyl-C(CH 3 ) 3 ), 1-pentyl (n-pentyl, -CH 2 CH 2 CH 2 CH 2 CH 3 ), 2-pentyl (-CH(CH 3 )CH 2 CH 2 CH 3 ), 3-pentyl (-CH(CH 2 CH 3 ) 2 ), 2-methyl-2-butyl (-C(CH 3 ) 2 CH 2 CH 3 ), 3-methyl-2-butyl (-CH(CH 3 )CH(CH 3 ) 2 ), 3-methyl-1-butyl (-CH 2 CH 2 CH(CH 3 ) 2 ), 2-methyl-1-butyl (-CH 2 CH(CH 3 )CH 2 CH 3 ), 1-hexyl (-CH 2 CH 2 CH 2 CH 2 CH 2 CH 3 ), 2-hexyl (-CH(CH 3 )CH 2 CH 2 CH 2 CH 3 ), 3-hexyl(-CH(CH 2 CH 3 )(CH 2 CH 2 CH 3 )), 2-methyl-2-pentyl (-C(CH 3 ) 2 CH 2 CH 2 CH 3 ), 3-methyl-2-pentyl (-CH(CH 3 )CH(CH 3 )CH 2 CH 3 ), 4-methyl-2-pentyl (-CH(CH 3 )CH 2 CH(CH 3 ) 2 ), 3-methyl-3-pentyl (-C(CH 3 )(CH 2 CH 3 ) 2 ), 2-methyl-3-pentyl (-CH(CH 2 CH 3 )CH(CH 3 ) 2 ), 2,3-dimethyl-2-butyl(-C(CH 3 ) 2 CH(CH 3 ) 2 ), 3,3-dimethyl-2-butyl(-CH(CH 3 )C(CH 3 ) 3 , 1-heptyl, 1-octyl and the like.

[0093] As used herein, the term "alkylene" refers to any length of 1 to 12 carbon atoms (C 1 ~C 12 ), wherein the alkylene radical is optionally substituted independently with one or more substituents described below. In another embodiment, the alkylene radical has 1-8 carbon atoms (C 1 ~C 8 ), or 1 to 6 carbon atoms (C 1 ~C 6 ). As an alkylene group, methylene (-CH 2 -), ethylene (-CH 2 CH 2 -), propylene (-CH 2 CH 2 CH 2 -) and the like, but are not limited to these.

[0094] The term "alkenyl" means that at least one site of unsaturation, i.e. carbon-carbon sp 2 2 to 8 carbon atoms with double bonds (C 2 ~C 8), alkenyl radicals are optionally substituted independently with one or more substituents described herein, including radicals having “cis” and “trans” orientations, or alternatively “E” and “Z” orientations. Examples include ethynylenyl or vinyl (-CH=CH 2 ), allyl (-CH 2 CH=CH 2 ) and the like, but are not limited to these.

[0095] The term "alkenylene" means at least one site of unsaturation, i.e. carbon-carbon sp 2 2 to 8 carbon atoms with double bonds (C 2 ~C 8 ), wherein alkenylene radicals are optionally substituted independently with one or more substituents described herein, including radicals having “cis” and “trans” orientations, or alternatively “E” and “Z” orientations. Examples are ethynylene or vinylene (-CH=CH-), allyl (-CH 2 CH=CH-) and the like, but are not limited to these.

[0096] The term "alkynyl" refers to any length of 2 to 8 carbon atoms (C 2 ~C 8 ), wherein the alkynyl radical is optionally substituted independently with one or more substituents described herein. Examples include ethynyl (-C≡CH), propynyl (propargyl, -CH 2 C≡CH) and the like, but are not limited to

[0097] The term "alkynylene" refers to any length of 2 to 8 carbon atoms (C 2 ~C 8 ), wherein the alkynylene radical is optionally substituted independently with one or more substituents described herein. Examples include ethylene (-C≡C-), propynylene (propargylene, -CH 2 C≡C-) and the like, but are not limited to these.

[0098] The terms "carbocycle", "carbocyclyl", "carbocyclic ring" and "cycloalkyl" refer to 3 to 12 carbon atoms (C 3 ~C 12 ) or a monovalent non-aromatic saturated or partially unsaturated ring having 7 to 12 carbon atoms as a bicyclic ring. Bicyclic carbocycles having 7 to 12 atoms can be arranged, for example, as bicyclo[4,5], [5,5], [5,6], or [6,6] systems; It can be arranged as a crosslinked system such as naan. Spiro moieties are also included within the scope of this definition. Examples of monocyclic carbocycles include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclo Examples include, but are not limited to, dodecyl and the like. Carbocyclyl groups are optionally substituted independently with one or more substituents described herein.

[0099] "Aryl" means 6 to 20 carbon atoms (C 6 ~C 20 ) means a monovalent aromatic hydrocarbon radical. Some aryl groups are represented as "Ar" in exemplary structures. Aryl includes bicyclic radicals comprising an aromatic ring fused to a saturated, partially unsaturated ring, or aromatic carbocyclic ring. Typical aryl groups include, but are not limited to, radicals derived from benzene (phenyl), substituted benzene, naphthalene, anthracene, biphenyl, indenyl, indanyl, 1,2-dihydronaphthalene, 1,2,3,4-tetrahydronaphthyl, and the like. Aryl groups are optionally substituted independently with one or more substituents described herein.

[0100] "Arylene" refers to 6 to 20 carbon atoms (C 6 ~C 20 ) means a divalent aromatic hydrocarbon radical. Some arylene groups are represented as "Ar" in exemplary structures. Arylene includes bicyclic radicals comprising an aromatic ring fused to a saturated, partially unsaturated ring, or aromatic carbocyclic ring. Typical arylene groups include, but are not limited to, radicals derived from benzene (phenylene), substituted benzene, naphthalene, anthracene, biphenylene, indenylene, indanylene, 1,2-dihydronaphthalene, 1,2,3,4-tetrahydronaphthyl, and the like. Arylene groups are optionally substituted with one or more substituents described herein.

[0101] The terms "heterocycle," "heterocycle," and "heterocyclic ring" are used interchangeably herein and refer to a saturated or partially unsaturated (i.e., having one or more double and / or triple bonds in the ring) carbocyclic radical of 3 to about 20 ring atoms, at least one ring atom being a heteroatom selected from nitrogen, oxygen, phosphorus, and sulfur, the remaining ring atoms being C, and one or more ring atoms optionally being independently substituted with one or more substituents described below. The heterocycle may be a 3-7 membered (2-6 carbon atoms and 1-4 heteroatoms selected from N, O, P, and S) monocyclic or a 7-10 membered (4-9 carbon atoms and 1-6 heteroatoms selected from N, O, P, and S) bicyclic ring, for example, a bicyclo[4,5], [5,5], [5,6], or [6,6] system. Heterocycles are described in Paquette, Leo A; "Principles of Modern Heterocyclic Chemistry" (W.A. Benjamin, New York, 1968), especially Chapters 1, 3, 4, 6, 7, 9; 28; and J. Am. Chem. Soc. (1960) 82:5566. "Heterocycle" also includes radicals where heterocycle radicals are fused with saturated, partially unsaturated rings, or aromatic carbocyclic or heterocyclic rings. Heterocyclic rings include, for example, morpholin-4-yl, piperidin-1-yl, piperazinyl, piperazin-4-yl-2-one, piperazin-4-yl-3-one, pyrrolidin-1-yl, thiomorpholin-4-yl, S-dioxothiomorpholin-4-yl, azocan-1-yl, azetidin-1-yl, octahydropyrido[1,2-a]pyrazine- 2-yl, [1,4]diazepan-1-yl, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, homopiperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, Oxazepinyl, diazepinyl, thiazepinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolinylimidazolinyl, imidazolinyl, 3-azabicyclo[3.1.0]hexanyl , 3-azabicyclo[4.1.0]heptanyl, azabicyclo[2.2.2]hexanyl, 3H-indolylquinolidinyl, and N-pyridyl urea. Spiro moieties are also included within the scope of this definition. Examples of heterocyclic groups in which 2 ring atoms are substituted with oxo (=O) moieties are pyrimidinonyl and 1,1-dioxo-thiomorpholinyl. The heterocyclic groups herein are optionally substituted independently with one or more substituents described herein.

[0102] The term "heteroaryl" refers to a 5-, 6-, or 7-membered monovalent aromatic radical, including fused ring systems of 5-20 atoms, at least one of which is aromatic, containing one or more heteroatoms independently selected from nitrogen, oxygen, and sulfur. Examples of heteroaryl groups include pyridinyl (including for example 2-hydroxypyridinyl), imidazolyl, imidazopyridinyl, 1-methyl-1H-benzo[d]imidazole, [1,2,4]triazolo[1,5-a]pyridine, pyrimidinyl (including for example 4-hydroxypyrimidinyl), pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, iso xazolyl, thiazolyl, oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, thiadiazolyl, furazanyl, benzo furazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl. Heteroaryl groups are optionally substituted independently with one or more substituents described herein.

[0103] A heterocycle or heteroaryl group may be carbon (carbon-bonded) or nitrogen (nitrogen-bonded) bonded, where possible. By way of example, without limitation, a carbon-bonded heterocycle or heteroaryl may be at the 2, 3, 4, 5, or 6 positions of pyridine, the 3, 4, 5, or 6 positions of pyridazine, the 2, 4, 5, or 6 positions of pyrazine, the 2, 3, 5, or 6 positions of pyrazine, the 2, 3, 4, or 5 positions of furan, tetrahydrofuran, thiofuran, thiophene, pyrrole, or tetrahydropyrrole, oxazole, imidazole, or thiazole. 2, 4, or 5; isoxazole, pyrazole, or isothiazole; 3, 4, or 5; aziridine, 2, 3, or 4; azetidine, 2, 3, 4, 5, 6, 7, or 8;

[0104] By way of example, but not limitation, nitrogen-bonded heterocycles or heteroaryls include aziridine, azetidine, pyrrole, pyrrolidine, 2-pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline, 3-imidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline, piperidine, piperazine, indole, indoline, 1H-indazole at position 1, isoindazole. It is attached at the 2-position of dol or isoindoline, the 4-position of morpholine, and the 9-position of carbazole or β-carboline.

[0105] The term "chiral" refers to molecules which have the property of non-superimposability of their mirror image partner, while the term "achiral" refers to molecules which are superimposable on their mirror image partner.

[0106] The term "stereoisomers" refers to compounds that have identical chemical constitution but differ with respect to the arrangement of the atoms or groups in space.

[0107] "Diastereomer" refers to a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical properties such as melting points, boiling points, spectral properties, and reactivities. Mixtures of diastereomers may separate under high resolution analytical procedures such as electrophoresis and chromatography.

[0108] "Enantiomers" refer to two stereoisomers of a compound that are non-superimposable mirror images of each other.

[0109] Stereochemical definitions and conventions used herein generally follow S.P. Parker, Ed., "McGraw-Hill Dictionary of Chemical Terms" (1984), McGraw-Hill Book Company (New York) and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds" (1994), John Wiley & Sons, Inc., New York. Many organic compounds exist in optically active forms, ie, they have the ability to rotate the plane of plane-polarized light. In describing optically active compounds, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule about its chiral center. The prefixes d and l or (+) and (-) are used to designate the indication of rotation of plane-polarized light by the compound, with (-) or 1 meaning the compound is levorotatory. Compounds prefixed with (+) or d are dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. A particular stereoisomer may also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is termed a racemic mixture or racemate, which can occur when there is no stereoselectivity or stereospecificity in a chemical reaction or process. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomeric species devoid of optical activity.

[0110] Other terms, definitions and abbreviations herein include the following. 6-maleimidocaproyl (“MC”); maleimidopropanoyl (“MP”); valine-citrulline (“val-cit” or “vc”), alanine-phenylalanine (“ala-phe”), p-aminobenzyl (“PAB”) and p-aminobenzyloxycarbonyl (“PABC”); A1 18C (EU numbering) = A121C (sequential numbering) = A114C (Kabat numbering) of heavy chain K149C (Kabat numbering) of light chain. Additional definitions and abbreviations are provided herein.

[0111] II. Chemical degradation inducers A chemical degradation inducer (CIDE) molecule can be conjugated to the antibody to form an "Ab-CIDE" conjugate. The antibody is conjugated to CIDE (“D”) via a linker (L1), which includes a ubiquitin E3 ligase groug (“E3LB”), a linker (“L2”) and a protein binding group (“PB”). The general formula for Ab-CIDE molecules is: Ab-(L1-D) p L2 is the linker covalently attached to E3LB and PB; PB is the protein attachment group covalently attached to L2; Ab is the antibody covalently attached to L1; L1 is the linker covalently attached to Ab and D; The variable p reflects that an antibody can be linked to one or more L1-D groups. In one embodiment, p is 1-8. In another embodiment, p is about two.

[0112] The following section describes the components that make up Ab-CIDE. To obtain an Ab-CIDE with potent potency and desirable therapeutic index, the following ingredients are provided.

[0113] 1. Antibody (Ab) As described herein, antibodies, such as monoclonal antibodies (mAbs), are used to deliver CIDE to target cells, such as cells expressing the specific protein targeted by the antibody. The antibody portion of Ab-CIDE can target antigen-expressing cells, whereby antigen-specific Ab-CIDE is delivered intracellularly, typically by endocytosis, to target cells. Ab-CIDE containing antibodies to antigens not found on the cell surface may result in less specific intracellular delivery of the CIDE portion into the cell, although Ab-CIDE may still be pinocytosed. The Ab-CIDEs and their methods of use described herein advantageously take advantage of cell surface antibody recognition and / or endocytosis of Ab-CIDEs to deliver CIDE moieties intracellularly.

[0114] a human antibody In certain embodiments, the antibodies provided herein are human antibodies. Human antibodies can be produced using various techniques known in the art. Human antibodies are generally described in van Dijk and van de Winkel, "Curr. Opin. Pharmacol." 5:368-74 (2001) and Lonberg, "Curr. Opin. Immunol." 20:450-459 (2008).

[0115] Human antibodies may be prepared by administering an immunogen to an intact human antibody or a transgenic animal that has been modified to produce an intact antibody having human variable regions in response to antigenic challenge. Such animals typically contain all or part of the human immunoglobulin loci either replacing the endogenous immunoglobulin loci or existing extrachromosomally or randomly integrated into the animal's chromosomes. In such transgenic mice, the exogenous immunoglobulin loci are generally inactivated. For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech. 23:1117-1125 (2005). Also, for example, U.S. Patent Nos. 6,075,181 and 6,150,584 describing XENOMOUSE(TM) technology; U.S. Patent No. 5,770,429 describing HuMab(R) technology; U.S. Patent No. 7,041,870 describing K-M MOUSE(R) technology; See also 61900. Human variable regions from intact antibodies generated by such animals can be further modified, for example, by combining with a different human constant region.

[0116] Human antibodies can also be produced by hybridoma-based methods. Human myeloma and mouse-human xenomyeloma cell lines have been described for the production of human monoclonal antibodies. (See, e.g., Kozbor, J. Immunol. 133:3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., J. Immunol. 147:86 (1991)). Human antibodies generated via bridoma technology are also described in Li et al., Proc. Natl. Acad. Sci. USA 103:3557-3562 (2006). Additional methods include, for example, U.S. Pat. No. 7,189,826 (describing the production of monoclonal human IgM antibodies from hybridoma cell lines), and Ni, “Xiandai Mianyixue” 26:4:265-268 (2006) (describing human-human hybridomas). Human hybridoma technology (trioma technology) is also described in Vollmers and Brandlein, "Histology and Histopathology" 20(3):927-937 (2005) and Vollmers and Brandlein, "Methods and Findings in Experimental and Clinical Pharmacology" 27(3):185-91 (2005).

[0117] Human antibodies can also be generated by isolating Fv clone variable domain sequences selected from human-derived phage display libraries. Such variable domain sequences can then be combined with the desired human constant domain. Techniques for selecting human antibodies from antibody libraries are described below.

[0118] b. library-derived antibody Antibodies for use in Ab-CIDE can be isolated by screening combinatorial libraries for antibodies with the desired activity(s). For example, various methods are known in the art for generating phage display libraries and screening such libraries for antibodies possessing desired binding characteristics. Such methods are reviewed, for example, in Hoogenboom et al., Methods in Molecular Biology 178:1-37 (O'Brien et al., eds., Human Press, Totowa, NJ, 2001) and are described further below: e.g., McCafferty et al., Nature 348:552-554; Clackson et al., Nature 352:624-628. 222:581-597 (1992); Marks and Bradbury, Methods in Molecular Biology 248:161-175 (Lo et al., Human Press, Totowa, NJ, 2003); Sidhu et al., J. Mol. Biol. 338:2:29. 9-310 (2004); Lee et al., J. Mol. Biol. 340:5:1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. Volume 1-2, pp. 119-132 (2004).

[0119] In a particular phage display method, the VH and VL gene repertoires can be separately cloned by polymerase chain reaction (PCR), randomly recombined in a phage library, and then screened for antigen-binding phage as described in Winter et al., Ann. Rev. Immunol. 12:433-455 (1994). Phage typically display antibody fragments, either as single-chain Fv (scFv) fragments, or as Fab fragments. Libraries from immunization sources give high affinity antibodies to the immunogen without the need to construct hybridomas. Alternatively, the naive repertoire can be cloned (e.g., from humans) to provide a single antibody source against a wide range of non-self and self-antigens without any immunization, as described by Griffiths et al., EMBO J, 12:725-734 (1993). Finally, naive libraries can also be generated synthetically by cloning unrearranged V gene segments from stem cells, using PCR primers containing random sequences to encode the highly variable CDR3 region, and achieving rearrangement in vitro as described in Hoogenboom and Winter, J. Mol. Biol. 227:381-388 (1992). Patent publications describing human antibody phage libraries include, for example: U.S. Patent No. 5,750,373, and U.S. Patent Application Publications Nos. 2005 / 0079574, 2005 / 0119455, 2005 / 0266000, 2007 / 0117126, 2007 / 0160598; 2007 / 0237764, 2007 / 0292936, and 2009 / 0002360.

[0120] Antibodies or antibody fragments isolated from human antibody libraries are considered human antibodies or human antibody fragments herein.

[0121] c. Chimeric and humanized antibodies In certain embodiments, the antibodies provided herein are chimeric antibodies. Certain chimeric antibodies are described, for example, in US Pat. No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984). In one example, a chimeric antibody comprises a non-human variable region (eg, a variable region derived from a non-human primate such as mouse, rat, hamster, rabbit, or monkey) and a human constant region. In a further example, chimeric antibodies are "class-switched" antibodies in which the class or subclass has been altered from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0122] In certain embodiments, a chimeric antibody is a humanized antibody. Typically, non-human antibodies are humanized to reduce their immunogenicity to humans while retaining the specificity and affinity of the parent non-human antibody. Typically, a humanized antibody comprises one or more variable domains in which the HVRs, eg CDRs (or portions thereof) are derived from non-human antibodies and the FRs (or portions thereof) are derived from human antibody sequences. A humanized antibody optionally also will comprise at least a portion of a human constant region. In some embodiments, some FR residues in the humanized antibody are replaced with corresponding residues from a non-human antibody (e.g., the antibody from which the HVR residues are derived), e.g., to restore or improve antibody specificity or affinity.

[0123] Humanized antibodies and methods for making them are reviewed, for example, by Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008) and further described in: for example, Riechmann et al. : 10029-10033 (1989); U.S. Patent Nos. 5,821,337, 7,527,791, 6,982,321 and 7,087,409; Kashmiri et al., Methods 36:25-34 (2005) (describing specificity determining region (CDR) grafting); Padlan, Mol. Immunol.", 28:489-498 (1991) (describing "Resurfacing"); Dall'Acqua et al., "Methods", 36:43-60 (2005) (describing "FR Shuffle"); 83:252-260 (2000) (describes FR Shuffle's "guided selection approach").

[0124] Framework regions that can be used for humanization include, but are not limited to, those selected using the "best fit" method (see, e.g., Sims et al., J. Immunol., 151:2296 (1993)); :4285 (1992); and Presta et al., J. Immunol. 151:2623 (1993)); human mature (somatically mutated) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); Framework regions derived from screening (see, for example, Baca et al., J. Biol. Chem. 272:10678-10684 (1997) and Rosok et al., J. Biol. Chem. 271:22611-22618 (1996)).

[0125] d. Multispecific antibodies In certain embodiments, antibodies provided herein are multispecific antibodies, eg, bispecific antibodies. As used herein, the term "multispecific antibody" encompasses antibodies comprising antigen-binding domains that have multiple epitopic specificities (i.e., can bind two or more different epitopes on one biological molecule, or can bind epitopes on two or more different molecules).

[0126] In some embodiments, multispecific antibodies are monoclonal antibodies that have binding specificities for at least two different antigen binding sites (such as bispecific antibodies). In some embodiments, the first antigen binding domain and the second antigen binding domain of a multispecific antibody may bind two epitopes within one and the same molecule (intramolecular binding). For example, a first antigen binding domain and a second antigen binding domain of a multispecific antibody may bind to two different epitopes on the same protein molecule. In certain embodiments, the two different epitopes bound by a multispecific antibody are usually epitopes that are not bound simultaneously by a single monospecific antibody (such as a conventional antibody) or a single variable domain of an immunoglobulin. In some embodiments, the first antigen-binding domain and the second antigen-binding domain of a multispecific antibody may bind epitopes located within two separate molecules (intramolecular binding). For example, a first antigen binding domain of a multispecific antibody may bind to one epitope on one protein molecule, while a second antigen binding domain of a multispecific antibody may bind to another epitope on a different protein molecule, thereby bridging the two molecules.

[0127] In some embodiments, an antigen binding domain of a multispecific antibody (such as a bispecific antibody) comprises two VH / VL units, a first VH / VL unit binding a first epitope and a second VH / VL unit binding a second epitope, each VH / VL unit comprising a heavy chain variable domain (VH) and a light chain variable domain (VL). Such multispecific antibodies include, but are not limited to, full-length antibodies, antibodies with two or more VL and VH domains, and antibody fragments (such as covalently or non-covalently linked Fabs, Fvs, dsFvs, scFvs, diabodies, bispecific diabodies, and triabodies). A VH / VL unit that further comprises at least a portion of a heavy chain variable region and / or at least a portion of a light chain variable region may also be referred to as an "arm" or "hemimer" or "half-antibody." In some embodiments, the hemimer comprises a portion of the heavy chain variable region sufficient to allow an intramolecular disulfide bond to form with the second hemimer. In some embodiments, the hemimer contains knob or hole mutations that allow heterodimerization with, for example, a second hemimer or half-antibody that contains complementary hole or knob mutations. Knob and hole mutations are discussed further below.

[0128] In certain embodiments, multispecific antibodies provided herein can be bispecific antibodies. As used herein, the term "bispecific antibody" refers to a multispecific antibody that is capable of binding two different epitopes on one molecule or that contains an antigen binding domain capable of binding epitopes on two different molecules. Bispecific antibodies are also referred to herein as having "bispecificity" or being "bispecific." Exemplary bispecific antibodies can bind both the protein and any other antigen. In certain embodiments, one of the binding specificities is for a protein and the other is for CD3. See, for example, US Pat. No. 5,821,337. In certain embodiments, bispecific antibodies may bind to two different epitopes on the same protein molecule. In certain embodiments, bispecific antibodies can bind to two different epitopes on two different protein molecules. Bispecific antibodies can also be used to localize cytotoxic agents to cells which express the protein. Bispecific antibodies can be prepared as full length antibodies or antibody fragments.

[0129] Techniques for making multispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy-light chain pairs with different specificities (see Milstein and Cuello, Nature 305:537 (1983), WO 93 / 08829, and Traunecker et al. EMBO J. 10:3655 (1991)), and knob-in-hole. (see, e.g., U.S. Patent No. 5,731,168), WO 2009 / 089004, U.S. Patent Application Publication Nos. 2009 / 0182127, 2011 / 0287009, Marvin and Zhu, Acta Pharmacol. Sin. (2005) 26(6):649-658, and Kontermann (2005). 26:1-9), Acta Pharmacol. Sin. As used herein, the term "knob-in-to-hole" or "KnH" technology refers to a technique that directs the pairing of two polypeptides in vitro or in vivo by introducing a bulge (knob) in one polypeptide and a cavity (hole) in the other at the interface where they interact. For example, KnH has been introduced at the Fc:Fc binding interface, the CL:CH1 interface, or the VH / VL interface of an antibody (e.g., U.S. Patent Application Publication No. 2011 / 0287009, U.S. Patent Application Publication No. 2007 / 0178552, WO 96 / 027011, WO 98 / 050431, Zhu et al., 1997, Protein Science 6:78). 1-788, and WO 2012 / 106587). In some embodiments, KnH drives pairing of two different heavy chains during the production of multispecific antibodies. For example, multispecific antibodies that have KnH within their Fc regions may further comprise a single variable domain linked to each Fc region, or may further comprise different heavy chain variable domains paired with similar or different light chain variable domains. KnH technology can also be used to pair two different receptor extracellular domains together, or any other polypeptide sequences containing different target recognition sequences (e.g., including affibodies, peptibodies, and other Fc fusions).

[0130] As used herein, the term "knob mutation" refers to a mutation that introduces a bulge (knob) into a polypeptide at the interface where the polypeptide interacts with another polypeptide. In some embodiments, the other polypeptide has a hole mutation.

[0131] As used herein, the term "hole mutation" refers to a mutation that introduces a cavity (hole) into a polypeptide at the interface where the polypeptide interacts with another polypeptide. In some embodiments, the other polypeptide has a knob mutation.

[0132] A "protuberance" refers to at least one amino acid side chain that protrudes from the interface of the first polypeptide and thus can be positioned within the compensating cavity of the adjacent interface (i.e., the interface of the second polypeptide), e.g., to stabilize heteromultimers, thereby favoring heteromultimerization over homomultimerization. Protuberances can exist in the original interface or can be introduced synthetically (eg, by modifying the nucleic acid encoding the interface). In some embodiments, the nucleic acid encoding the interface of the first polypeptide is altered to encode a protrusion. To accomplish this, the nucleic acid encoding at least one "original" amino acid residue at the interface of the first polypeptide is replaced with nucleic acid encoding at least one "import" amino acid residue having a larger side chain volume than the original amino acid residue. It is understood that there may be more than one original residue and corresponding imported residue. Side chain volumes for various amino residues are shown, for example, in Table 1 of US Patent Application Publication No. 2011 / 0287009. A mutation that introduces a "protrusion" may be referred to as a "knob mutation."

[0133] In some embodiments, the import residue for ridge formation is a naturally occurring amino acid residue selected from arginine (R), phenylalanine (F), tyrosine (Y), and tryptophan (W). In some embodiments, the import residue is tryptophan or tyrosine. In some embodiments, the original residues for bulge formation such as alanine, asparagine, aspartic acid, glycine, serine, threonine, or valine have small side chain volumes.

[0134] A "cavity" refers to at least one amino acid side chain that is recessed from the interface of the second polypeptide and thus accommodates a corresponding ridge on the interface of the adjacent first polypeptide. Cavities can exist in the original interface or can be introduced synthetically (eg, by modifying the nucleic acid encoding the interface). In some embodiments, the nucleic acid encoding the interface of the second polypeptide is altered to encode a cavity. To accomplish this, the nucleic acid encoding at least one "original" amino acid residue at the interface of the second polypeptide is replaced with DNA encoding at least one "import" amino acid residue having a smaller side chain volume than the original amino acid residue. It is understood that there may be more than one original residue and corresponding imported residue. In some embodiments, import residues for cavity formation are naturally occurring amino acid residues selected from alanine (A), serine (S), threonine (T), and valine (V). In some embodiments, the import residue is serine, alanine, or threonine. In some embodiments, original residues for cavity formation, such as tyrosine, arginine, phenylalanine, or tryptophan, have large side chain volumes. A mutation that introduces a "cavity" may be referred to as a "hole mutation."

[0135] A protuberance is "positionable" within a cavity, which means that the spatial location of the protuberance and cavity at the interface of the first and second polypeptides, respectively, and the size of the protuberance and cavity are such that the protuberance can be positioned within the cavity without significantly disturbing the normal association of the first and second polypeptides at the interface. Protrusions such as Tyr, Phe, and Trp typically do not extend perpendicularly from the interfacial axis and do not have a preferred conformation, and alignment of protuberances with corresponding cavities can in some cases rely on modeling the protuberance / cavity pair based on three-dimensional structures such as those obtained by X-ray crystallography or nuclear magnetic resonance (NMR). This can be accomplished using techniques widely accepted in the art.

[0136] In some embodiments, the knob mutation within the IgG1 constant region is T366W (EU numbering). In some embodiments, the Hall mutation within the IgG1 constant region comprises one or more mutations selected from T366S, L368A, and Y407V (EU numbering). In some embodiments, Hall mutations within the IgG1 constant region comprise T366S, L368A, and Y407V (EU numbering).

[0137] In some embodiments, the knob mutation within the IgG4 constant region is T366W (EU numbering). In some embodiments, the Hall mutation within the IgG4 constant region comprises one or more mutations selected from T366S, L368A, and Y407V (EU numbering). In some embodiments, Hall mutations within the IgG4 constant region comprise T366S, L368A, and Y407V (EU numbering).

[0138] Multispecific antibodies are engineered by electrostatic steering to create antibody Fc-heterodimeric molecules (WO 2009 / 089004 A1); cross-linking two or more antibodies or fragments (see, e.g., U.S. Pat. No. 4,676,980, and Brennan et al., Science, 229:81 (1985)); use of leucine zippers to generate bispecific antibodies. (See, e.g., Kostelny et al., J. Immunol., 148(5):1547-1553 (1992)); the use of "diabody" technology to generate bispecific antibody fragments (e.g., Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993)); Dimers can be used (see, eg, Gruber et al., J. Immunol., 152:5368 (1994)); and can be made, eg, as described in Tutt et al., J. Immunol. 147:60 (1991).

[0139] Also included herein are engineered antibodies with three or more functional antigen-binding sites, including "octopus antibodies" or "dual variable domain immunoglobulins" (DVDs) (e.g., U.S. Patent Application Publication No. 2006 / 0025576 A1, and Wu et al., Nature Biotechnology (2007)). Antibodies or fragments herein also include "Dual Acting FAbs" or "DAFs" that contain an antigen binding site that binds a target protein as well as another, different antigen (see, eg, US2008 / 0069820).

[0140] e.Antibody fragment In certain embodiments, the antibodies provided herein are antibody fragments. Antibody fragments include Fab, Fab', Fab'-SH, F(ab') 2 , Fv, and scFv fragments, as well as other fragments described below. For a review of specific antibody fragments, see Hudson et al., Nat. Med. 9:129-134 (2003). For a review of scFv fragments, see, eg, Pluckthun, in The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore, eds. (Springer-Verlag, New York), pp. 269-315 (1994); see also WO 93 / 16185, US Pat. Please refer to Fabs and F(ab') containing salvage receptor binding epitope residues and increased in vivo half-lives 2 See US Pat. No. 5,869,046 for a description of fragments.

[0141] Diabodies are antibody fragments with two antigen-binding sites that can be bivalent or bispecific. See, for example, EP 404,097, International Publication No. WO1993 / 01161, Hudson et al., Nat. Med. 9:129-134 (2003) and Hollinger et al., Proc. Natl. Acad. Trispecific and tetraspecific antibodies are also described in Hudson et al., Nat. Med. 9:129-134 (2003).

[0142] Single domain antibodies are antibody fragments that contain all or part of the heavy chain variable domain or all or part of the light chain variable domain of an antibody. In certain embodiments, a single domain antibody is a human single domain antibody (Domantis, Inc., Waltham, Mass.; see, eg, US Pat. No. 6,248,516 B1).

[0143] Antibody fragments can be produced by a variety of techniques, including, but not limited to, proteolytic digestion of intact antibodies and production by recombinant host cells (e.g., E. coli or phage), as described herein.

[0144] f.Antibody variants In certain embodiments, amino acid sequence variants of the antibodies provided herein are contemplated. For example, it may be desirable to improve the binding affinity and / or other biological properties of an antibody. Amino acid sequence variants of the antibody may be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody, or by peptide synthesis. Such modifications include, for example, deletions from, and / or insertions into, and / or substitutions of residues within the amino acid sequences of the antibody. Any combination of deletion, insertion, and substitution can be made to arrive at the final construct, provided that the final construct possesses the desired properties, eg, antigen binding.

[0145] g. Recombinant Methods and Compositions Antibodies may be produced, for example, using recombinant methods and compositions described in US Pat. No. 4,816,567. In one embodiment, an isolated nucleic acid encoding an antibody described herein is provided. Such nucleic acids can encode amino acid sequences comprising the VL and / or the VH of an antibody (eg, the light and / or heavy chains of an antibody). In further embodiments, one or more vectors (eg, expression vectors) containing such nucleic acids are provided. In further embodiments, host cells containing such nucleic acids are provided. In one such embodiment, the host cell comprises (e.g., has been transformed with): (1) a vector comprising a nucleic acid encoding an amino acid sequence comprising the VL of the antibody and an amino acid sequence comprising the VH of the antibody, or (2) a first vector comprising a nucleic acid encoding an amino acid sequence comprising the VL of the antibody, and a second vector comprising (e.g., transformed with) a nucleic acid encoding an amino acid sequence comprising the VH of the antibody. In one embodiment, the host cell is a eukaryotic cell such as a Chinese Hamster Ovary (CHO) cell or a lymphocytic cell (eg Y0, NS0, Sp20 cells). In one embodiment, a method of producing an antibody is provided, the method comprising culturing a host cell containing a nucleic acid encoding the antibody described above under conditions suitable for expression of the antibody, and optionally recovering the antibody from the host cell (or host cell culture medium).

[0146] For recombinant production of antibodies, for example, nucleic acids encoding the antibodies described above are isolated and inserted into one or more vectors for further cloning and / or expression in a host cell. Such nucleic acids can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes capable of specifically binding to the genes encoding the heavy and light chains of the antibody).

[0147] Suitable host cells for cloning or expression of antibody-encoding vectors include prokaryotic or eukaryotic cells described herein. For example, antibodies may be produced in bacteria, particularly when glycosylation and effector functions are not required. See, eg, US Pat. Nos. 5,648,237, 5,789,199, and 5,840,523 for expression of antibody fragments and polypeptides in bacteria. (Charlton, "Methods in Molecular Biology", 248 (see also B.K.C.Lo, ed., Humana Press, Totowa, NJ, 2003), describing the expression of antibody fragments in E. coli, pp. 245-254) After expression, the antibody can be isolated from the bacterial cell paste in the soluble fraction and further purified.

[0148] In addition to prokaryotes, eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for antibody-encoding vectors, including fungal and yeast strains in which the glycosylation pathway has been "humanized" to produce antibodies with a partially or fully human glycosylation pattern. 22:1409-1414 (2004) and Li et al., Nat. Biotech. 24:210-215 (2006).

[0149] Suitable host cells for the expression of glycosylated antibodies are also derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant cells and insect cells. A number of baculovirus strains have been identified and may be used in combination with insect cells, particularly for transfection of Spodoptera frugiperda cells.

[0150] Plant cell cultures can also be used as hosts. See, for example, U.S. Patent Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (describing PLANTIBODIES™ technology for producing antibodies in transgenic plants).

[0151] Vertebrate cells may also be used as hosts. For example, mammalian cell lines adapted to grow in suspension may be useful. Non-limiting examples of useful mammalian host cell lines include the monkey kidney CV1 strain transformed with SV40 (COS-7); human embryonic kidney lines (e.g., 293 or 293 cells described in Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK); mouse Sertoli cells (e.g., Mather, "Biol. Reprod." 23:243-251 (1980); monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical carcinoma cells (HELA); Sci 383:44-68 (1982); MRC 5 cells; and FS4 cells.Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR-CHO cells (Urlaub et al., Proc. Natl. Acad. Sci. 2 / 0, etc. For a review of specific mammalian host cell lines suitable for antibody production, see, eg, Yazaki and Wu, Methods in Molecular Biology, 248 (B.K.C.Lo, ed., Human APress, Totowa, NJ) 255-268 (2003).

[0152] Referring now to antibody affinity, in embodiments the antibody binds to one or more tumor-associated antigens or cell surface receptors selected from (1)-(53):

[0153] (1) BMPR1B (bone morphogenetic protein receptor type IB, Genbank accession number NM_001203) ten Dijke, P. et al., Science 264(5155):101-104 (1994), Oncogene 14(11):1377-1382 (1997), WO2004063362 (claim 2), WO2003042661 (claim 12), U.S. Patent Application Publication No. 20031. 34790 A1 (pages 38-39), International Publication No. 2002102235 (claim 13, page 296), International Publication No. 2003055443 (91-92 pages), International Publication No. 200299122 (Example 2, pages 528-530), International Publication No. 2003029421 (claim 6), International Publication No. 2003024 392 (claim 2, Figure 112), WO200298358 (claim 1, page 183), WO200254940 (pages 100-101), WO200259377 (pages 349-350), WO200230268 (claim 27, page 376), WO200148204 (implementation e.g. Fig. 4) NP_001194 bone morphogenetic protein receptor, type IB / pid=NP_001194.1- Cross Reference: MIM:603248;NP_001194.1;AY065994

[0154] (2) E16 (LAT1, SLC7A5, Genbank accession number NM_003486) ``Biochem. Biophys. Res. Commun.'' 255(2), pp. 283-288 (1999); WO2004048938 (Example 2); WO2004032842 (Example IV); WO2003042661 (Claim 12); WO2003016475 (Claim 1); WO200278524 (Example 2); WO 200286443 (claim 27; pp. 222, 393); WO 2003003906 (claim 10; pp. 293); WO 200264798 (claim 33; pp. 93-95); WO 200014228 (claim 5; pp. 133-136); 224454 (Fig. 3); WO2003025138 (claim 12; page 150); NP_003477 solute carrier family 7 (cationic amino acid transporter, y+ system), member 5 / pid=NP_003477.3-homo sapiens Cross Reference: MIM:600182;NP_003477.3;NM_015923;NM_003486_1

[0155] (3) STEAP1 (Prostate 6 transmembrane epithelial antigen, Genbank accession number NM_012449) "Cancer Res." 61(15), pp. 5857-5860 (2001); Hubert, R.S. et al. (1999) "Proc. Natl. Acad. Sci. U.S.A." 96(25):14523-14528); (FIG. 1L); EP 1394274 (Example 11); WO2004016225 (Claim 2); WO2003042661 (Claim 12); No. (Figure 2); WO200289747 (Example 5; pages 618-619); WO2003022995 (Example 9; Figure 13A, Example 53; Page 173, Example 2; Figure 2A); NP_036581 Six transmembrane epithelial antigen of the prostate Cross Reference: MIM:604415;NP_036581.1;NM_012449_1

[0156] (4) 0772P (CA125, MUC16, Genbank accession number AF361486) "J.Biol.Chem." 276(29):27371-27375 (2001)); WO2004045553 (claim 14); WO200292836 (claim 6; Figure 12); 24140 (Example 16); U.S. Patent Application Publication No. 798959. Cross Reference: GI:34501467; AAK74120.3; AF361486_1

[0157] (5) MPF (MPF, MSLN, SMR, megakaryocyte enhancing factor, mesothelin, Genbank Accession No. NM_005823) Yamaguchi, N. et al. "Biol. Chem." 269(2), pp. 805-808 (1994), "Proc. Natl. Acad. Sci. Sci.U.S.A. 93(1):136-140 (1996), J.Biol.Chem. 270(37):21984-21990 (1995)); WO2003101283 (claim 14); 13; pp. 287-288); WO 2002101075 (claim 4; pp. 308-309); WO 200271928 (pp. 320-321); WO 9410312 (pp. 52-57);

[0158] (6) Napi2b (Napi3b, NAPI-3B, NPTIIb, SLC34A2, Solute Transporter Family 34 (Sodium Phosphate), Member 2, Type II Sodium-Dependent Phosphate Transporter 3b, Genbank Accession No. NM_006424) J. Biol. Chem. 277(22): 19665-19672 (2002), "Genomics" 62(2): 281-284 (1999), Feild, J.A. et al. (1999) "Biochem. Biophys. Res. Commun." 258(3): 578-582); 778 (Claim 2); EP 1394274 (Example 11); WO 2002102235 (Claim 13; 326 pages); Publication No. 200175177 (claim 24; pages 139-140); Cross Reference: MIM:604217;NP_006415.1;NM_006424_1

[0159] (7) Sema 5b (FLJ10372, KIAA1445, Mm.42015, SEMA5B, SEMAG, Semaphorin 5b Hlog, Sema domain, seven thrombospondin repeats (type 1 and type 1-like), transmembrane domain (TM), and short cytoplasmic domain, (Semaphorin) 5B, Genbank accession number AB040878) Nagase T. et al. (2000) "DNA Res." 7(2): pp. 143-150), International Publication No. 2004000997 (claim 1), International Publication No. 2003003984 (claim 1), International Publication No. 200206339 (claim 1, page 50), International Publication No. 200188133 (claims 1, 41-43, 48- 58 pages), International Publication No. 2003054152 (claim 20), International Publication No. 2003101400 (claim 11), Contract: Q9P283;EMBL;AB040878;BAA95969.1.Genew;HGNC:10737;

[0160] (8) PSCA hlg (2700050C12Rik, C530008O16Rik, RIKEN cDNA 2700050C12, RIKEN cDNA 2700050C12 gene, Genbank accession number AY358628); Ross et al. (2002) Cancer Res. 62:2546-2553; 9192 (claim 2); U.S. Patent Application Publication No. 2004044180 (claim 12); U.S. Patent Application Publication No. 2004044179 (claim 11); U.S. Patent Application Publication No. 2003096961 (claim 11); Patent Application Publication No. 2003206918 (Example 5); EP1347046 (Claim 1); WO2003025148 (Claim 20); Cross Reference: GI:37182378;AAQ88991.1;AY358628_1

[0161] (9) ETBR (endothelin type B receptor, Genbank accession number AY275463); Nakamuta M et al. "Biochem.Biophys.Res.Commun." 177, pp.34-39, 1991; Ogawa Y. et al. Chem.” 268, 3463-3470, 1993; Sakamoto A., Yanagisawa M. et al., “Biochem. Biophys. Res. Commun.” 178, pp. 656-663, 1991; "J. Cardiovasc. Pharmacol." 20, s1-S4, 1992; Tsutsumi M. et al. "Gene" 228, pp. 43-49, 1999; Strausberg R. L. et al. Metab." 82, pp. 3116-3123, 1997; Okamoto Y. et al. "Biol. Chem." 272, pp. 21589-21596, 1997; Verheij J.B. et al. 5, 180-185, 1997; Puffenberger E. G. et al., Cell 79, 1257-1266, 1994; Attie T. et al., Hum. Mol. Genet. 4, 2407-2409, 1995; 6; Amiel J. et al., ``Hum. Mol. Genet.'' 5, pp. 355-357, 1996; Hofstra R.M.W. et al., ``Nat. 7, pp. 115-124, 2001; Pingault V. et al. (2002) "Hum. Genet." 111, pp. 198-206; WO2004045516 (claim 1); WO2004048938 (example 2); WO 2003016475 (claim 1); WO 2003016475 (claim 1); WO 200261087 (Fig. 1); WO 2003016494 (Fig. 6); WO 2003025138 (claim 12; page 144); 351 (Claim 1; pp. 124-125); EP 522868 (Claim 8; Figure 2); WO 200177172 (Claim 1; pp. 297-299); ; International Publication No. 2004001004;

[0162] (10) MSG783 (RNF124, hypothetical protein FLJ20315, Genbank accession number M_017763); International Publication No. 2003104275 (claim 1); International Publication No. 2004046342 (Example 2); International Publication No. 2003042661 (claim 12); International Publication No. 2003083074 (claim 14, page 61); Section 2, Figure 93); WO200166689 (Example 6); Cross Reference: LocusID:54894;NP_060233.2;NM_017763_1

[0163] (11) STEAP2 (HGNC_8639, IPCA-1, PCANAP1, STAMP1, STEAP2, STMP, prostate cancer-associated gene 1, prostate cancer-associated protein 1, prostate 6-transmembrane epithelial antigen 2, 6-transmembrane prostate protein, Genbank accession number AF455138) 82(11): 1573-1582 (2002)); WO 2003087306, U.S. Patent Application Publication No. 2003064397 (claim 1, Figure 1); WO 200272596 (claim 13, pages 54-55); WO 200172962 (claim 1, Figure 4B); WO2003104270 (claim 11); WO2003104270 (claim 16); U.S. Publication No. 2004005598 (claim 22); WO2003042661 (claim 12); No. 22 (Claim 23, Figure 2); WO200216429 (Claim 12, Figure 10); Cross Reference: GI:22655488;AAN04080.1;AF455138_1

[0164] (12) TrpM4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptor potential cation channel, subfamily M, member 4, Genbank accession number NM_017636) U.S.A. 98(19): 10692-10697 (2001); Cell 109(3): 397-407 (2002); J. Biol. Chem. 278(33): 30813-30820 (2003)); WO2003143557 (claim 4); WO200040614 (claim 14, pages 100-103); WO200210382 (claim 1, Figure 9A); WO2003042661 (claim 12); WO200230268 (claim 27, page 391); 2003219806 (claim 4); WO200162794 (claim 14, Figures 1A-D); Cross Reference: MIM:606936;NP_060106.2;NM_017636_1

[0165] (13) CRIPTO (CR, CR1, CRGF, CRIPTO, TDGF1, teratocarcinoma-derived growth factor, Genbank accession number NP_003203 or NM_003212) Ciccodicola, A. et al. "EMBO J." 8(7): 1987-1991 (1989), "Am. J. Hum. Genet." 49(3): 555-565 (1991)); WO 200288170 (claim 2; pages 52-53); WO 2003024392 (claim 2; Figure 58); WO 200216413 (claim 1; pages 94-95, 105); WO 200222808 (claim 2; Figure 1); 4399 (Example 2; columns 17-18); US Pat. No. 5,792,616 (FIG. 2); Cross Reference: MIM:187395;NP_003203.1;NM_003212_1

[0166] (14) CD21 (CR2 (complement receptor 2) or C3DR (C3d / Epstein-Barr virus receptor) or Hs.73792, Genbank accession number M26004) Fujisaku et al. (1989) J. Biol. Chem. 264(4):2118-2125); Weis J.J. et al. Barel M. et al. "Mol. Immunol." 35, pp. 1025-1031, 1998; Weis J.J. et al. "Proc. Natl. Acad. Sci. WO2004045520 (Example 4); WO2004045520 (Example 4); WO9102536 (Figures 9.1 to 9.9); WO2004020595 (Claim 1); Acceptance: P20023;Q13866;Q14212;EMBL;M26004;AAA35786.1.

[0167] (15) CD79b (CD79B, CD79β, IGb (immunoglobulin-associated β), B29, Genbank accession number NM_000626 or 11038674) Sci.U.S.A. (2003) 100(7):4126-4131; "Blood" (2002) 100(9):3068-3076; Muller et al. (1992) Eur.J.Immunol. 22(6):1621-1625); WO2003087768, US2004101874 (claim 1, page 102); WO2003062401 (claim 9); WO200278524 (Example 2); US2002150573 (claim 5, page 15) WO 5644033; WO 2003048202 (claim 1, pages 306 and 309); WO 99 / 558658, US 6534482 (claim 13, Figure 17A / B); Cross Reference: MIM:147245;NP_000617.1;NM_000626_1

[0168] (16) FcRH2 (IFGP4, IRTA4, SPAP1A (SH2 domain containing phosphatase anchor protein 1a), SPAP1B, SPAP1C, Genbank accession number NM_030764, AY358130) "Genome Res." 13(10):2265-2270 (2003), "Immunogenetics" 54(2): 87-95 (2002), "Blood" 99(8): 2662-2669 (2002), "Proc.Natl.Acad.Sci.U.S.A."98(17):9772-977 7 (2001), Xu, M.J. et al. (2001) "Biochem. Biophys. Res. Commun." 280(3):768-775; WO2004016225 (claim 2); WO2003077836; WO2003097803 (claim 12); WO2003089624 (claim 25); Cross Reference: MIM:606509;NP_110391.2;NM_030764_1

[0169] (17) HER2 (ErbB2, Genbank accession number M11730) Coussens L. et al. "Science" (1985) 230(4730):1132-1139); Yamamoto T. et al. "Nature" 319, 230-234, 1986; Semba K. et al. U.S.A." 82, pp. 6497-6501, 1985; Swiercz J.M. et al. "J. Cell Biol." 165, pp. 869-880, 2004; Kuhns J. J. et al. Cho H.-S. et al. "Nature" 421, 756-760, 2003; Ehsani A. et al. (1993) "Genomics" 15, pp. 426-429; WO2004048938 (Example 2); WO2004027049 (Figure 1I); WO2004009622; WO2003081210; WO2003089904 (claim 9); WO2003016475 (claim 1); International Publication No. 2003008537 (claim 1); International Publication No. 2003055439 (claim 29; Figures 1A-B); International Publication No. 2003025228 (claim 37; Figure 5C); International Publication No. 200222636 (implementation Example 13; pages 95-107); International Publication No. 200212341 (claim 68; Figure 7); International Publication No. 200213847 (pages 71-74); 200153463 (claim 2; pages 41 to 46); International Publication No. 200141787 (page 15); International Publication No. 200044899 (claim 52; Figure 7); International Publication No. 200020579 (claim 3; Figure 2) US Patent No. 5869445 (claim 3; columns 31-38); International Publication No. 9630514 (claim 2; pages 56-61); European Patent No. 1439393 (claim 7); Claim 7); WO2004022709; WO200100244 (Example 3; Figure 4); Acceptance: P04626;EMBL;M11767;AAA35808.1.EMBL;M11761;AAA35808.1.

[0170] (18) NCA (CEACAM6, Genbank accession number M18728); Barnett T. et al. "Genomics" 3, pp. 59-66, 1988; Tawaragi Y. et al. "Biochem. Biophys. Res. Commun." 150, pp. 89-96, 1988; Strausberg R. L. et al. U.S.A." 99:16899-16903, 2002; WO2004063709; EP 1439393 (claim 7); WO2004044178 (Example 4); No. 2003042661 (claim 12); International Publication No. 200278524 (Example 2); International Publication No. 200286443 (claim 27; page 427); International Publication No. 200260317 (claim 2); Acceptance: P40199;Q14920;EMBL;M29541;AAA59915.1.EMBL;M18728;

[0171] (19) MDP (DPEP1, Genbank accession number BC017023) "Proc. Natl. Acad. Sci. U.S.A." 99(26): pp. 16899-16903 (2002)); page); Japanese Patent Application Publication No. 05003790 (Figs. 6 to 8); International Publication No. 9946284 (Fig. 9); Cross Reference: MIM:179780;AAH17023.1;BC017023_1

[0172] (20) IL20Ra (IL20Ra, ZCYTOR7, Genbank accession number AF184971); Clark H.F. et al., Genome Res. 13, 2265-2270, 2003; Mungall A.J. et al., Nature 425, 805-811, 2003; Blumberg H. et al., Cell 104, 9-19, 2001. 167, 3545-3549, 2001; Parrish-Novak J. et al., J. Biol. Chem. 277, 47517-47523, 2002; Pletnev S. et al. 2003) Biochemistry 42:12617-12624; Sheikh F. et al. (2004) J. Immunol. 172, 2006-2010; EP 1394274 (Example 11); 2004005320 (Example 5); International Publication No. 2003029262 (pages 74-75); International Publication No. 2003002717 (claim 2; page 63); International Publication No. 200222153 (pages 45-47); No. 2002042366 (pages 20-21); International Publication No. 200146261 (pages 57-59); International Publication No. 200146232 (pages 63-65); International Publication No. 9837193 (claim 1; pages 55-59); Contract: Q9UHF4;Q6UWA9;Q96SH8;EMBL;AF184971;AAF01320.1.

[0173] (21) Brevican (BCAN, BEHAB, Genbank accession number AF229053) Gary S.C. et al. "Gene" 256, pp. 139-147, 2000; Clark H.F. et al. "Genome Res." 13, pp. 2265-2270, 2003; Strausberg R.L. 16899-16903, 2002; U.S. Patent Application Publication No. 2003186372 (claim 11); U.S. Patent Application Publication No. 2003186373 (claim 11); U.S. Patent Application Publication No. 2003119131 (claim 1; Figure 52); U.S. Patent Application Publication No. 2003119122 (Claim 1; Figure 52); U.S. Patent Application Publication No. 2003119126 (Claim 1); U.S. Patent Application Publication No. 2003119121 (Claim 1; Figure 52); US Patent Application Publication No. 2003119129 (Claim 1); U.S. Patent Application Publication No. 2003119130 (Claim 1); U.S. Patent Application Publication No. 2003119128 (Claim 1; Figure 52); Publication No. 2003016475 (claim 1); WO200202634 (claim 1);

[0174] (22) EphB2R (DRT, ERK, Hek5, EPHT3, Tyro5, Genbank accession number NM_004442) Chan, J. and Watt, V.M. Oncogene 6(6):1057-1061 (1991) Oncogene 10(5):897-905 (1995) Annu. Rev. Neurosci. : 309-345 (1998), "Int. Rev. Cytol. 196: 177-244 (2000)); International Publication No. 2003042661 (claim 12); 41 pages); International Publication No. 2004065576 (claim 1); International Publication No. 2004020583 (claim 9); International Publication No. 2003004529 (pages 128 to 132); ); Cross Reference: MIM:600997;NP_004433.2;NM_004442_1

[0175] (23) ASLG659 (B7h, Genbank accession number AX092328) US Patent Application Publication No. 20040101899 (claim 2); WO2003104399 (claim 11); WO2004000221 (Figure 3); US Patent Application Publication No. 2003165504 (claim 1); Publication No. 2003124140 (Example 2); US Patent Application Publication No. 2003065143 (FIG. 60); WO 2002102235 (Claim 13; 299); No. 200210187 (claim 6; Figure 10); International Publication No. 200194641 (claim 12; Figure 7b); International Publication No. 200202624 (claim 13; Figures 1A-1B); Claim 54; pages 45-46); International Publication No. 200206317 (Example 2; pages 320-321, Claim 34; pages 321-322); International Publication No. 200271928 (pages 468-469); 200202587 (Example 1; Figure 1); International Publication No. 200140269 (Example 3; pp. 190-192); International Publication No. 200036107 (Example 2; pp. 205-207); Claim 12); International Publication No. 2003004989 (claim 1); International Publication No. 200271928 (pages 233-234, 452-453); International Publication No. 0116318;

[0176] (24) PSCA (prostate stem cell antigen precursor, Genbank accession number AJ297436) Natl. EP 1394274 (Example 11); US Patent Application Publication No. 2004018553 (claim 17); WO 2003008537 (claim 1); WO 200281646 (claim 1; page 164); 200140309 (Example 1; Figure 17); U.S. Patent Application Publication No. 2001055751 (Example 1; Figure 1b); WO 200032752 (Claim 18; Figure 1); No. (Claim 2; Figure 1B); Contract: O43653;EMBL;AF043498;AAC39607.1.

[0177] (25) GEDA (Genbank accession number AY260763); AAP14954 lipoma HMGIC fusion-partner-like protein / pid=AAP 14954.1-homo sapiens Species Homo sapiens (human) WO2003054152 (claim 20); WO2003000842 (claim 1); WO2003023013 (example 3, claim 20); US2003194704 (claim 45); Cross Reference: GI:30102449;AAP14954.1;AY260763_1

[0178] (26) BAFF-R (B cell activator receptor, BLyS receptor 3, BR3, Genbank accession number AF116456); BAFF receptor / pid=NP_443177.1-homo sapiens Thompson, J.S. et al. "Science" 293(5537), pp. 2108-2111 (2001); WO2004058309; WO2004011611; ); WO2003035846 (claim 70; pages 615-616); WO200294852 (columns 136-137); WO200238766 (claim 3; page 133); WO200224909 (example 3; Figure 3); Cross Reference: MIM:606269;NP_443177.1;NM_052945_1;AF132600

[0179] (27) CD22 (B cell receptor CD22-B isoform, BL-CAM, Lyb-8, Lyb8, SIGLEC-2, FLJ22814, Genbank accession number AK026467); Wilson et al. (1991) J.Exp.Med. 173:137-146; WO2003072036 (claim 1; Figure 1); Cross Reference: MIM:107266;NP_001762.1;NM_001771_1

[0180] (28) CD79a (B-cell specific protein that covalently interacts with CD79A, CD79α, immunoglobulin-associated alpha, Igbeta (CD79B) and forms surface complexes with IgM molecules, transducing signals involved in B-cell differentiation), pI: 4.84, MW: 25028 TM: 2[P] gene Chromosome: 19q13.2, Genbank Accession No. NP_001774.10) WO2003088808, U.S. Publication No. 20030228319; WO2003062401 (Claim 9); U.S. Publication No. 2002150573 (Claim 4, pages 13-14); WO9958658 (Claim 13, Figure 16); WO9207574 (Figure 1); U.S. Pat. No. 5,644,033; Ha et al. (1992) J. Immunol. 148(5):1526-1531; Mueller et al. (1992) Eur. J. Biochem. 22:1621-1625; Hashimoto et al. (1992) Clin. Exp. Immunol. 90(1): 141-146; Yu et al. (1992) J. Immunol. 148(2) pp. 633-637; Sakaguchi et al.

[0181] (29) CXCR5 (Burkitt's Lymphoma Receptor 1, a G protein-coupled receptor activated by the CXCL13 chemokine, functions in lymphocyte migration and humoral defense and plays a role in HIV-2 infection and possibly the development of AIDS, lymphoma, myeloma and leukemia); NP_001707.1) WO2004040000; WO2004015426; U.S. Publication No. 2003105292 (Example 2); U.S. Patent Application Publication No. 6555339 (Example 2); WO200261087 (Figure 1); 172830 (pages 12-13); International Publication No. 200022129 (Example 1, pages 152-153; Example 2, pages 254-256); International Publication No. 9928468 (claim 1, page 38); WO9217497 (claim 7, Figure 5); Dobner et al. (1992) Eur. J. Immunol. 22:2795-2799; Barella et al. (1995) Biochem. J. 309:773-779;

[0182] (30) HLA-DOB (beta subunit of MHC class II molecule (Ia antigen) that binds peptides and presents them to CD4+ T lymphocytes); 273 aa, pI: 6.56 MW: 30820 TM: 1 [P]Gene Chromosome: 6p21.3, Genbank Accession No. NP_002111.1) Tonnelle et al. (1985) EMBO J. 4(11):2839-2847; Jonsson et al. (1989) Immunogenetics 29(6):411-413; Beck et al. (1992) J. Mol. Biol. Sci USA 99:16899-16903; Servenius et al. (1987) J. Biol. Chem. 262:8759-8766; Beck et al. (1996) J. Mol. Biol. 658 (claim 13, Figure 15); U.S. Patent No. 6,153,408 (columns 35-38); U.S. Patent No. 5,976,551 (columns 168-170); U.S. Patent No. 6,011,146 (columns 145-146); "J. Biol. Chem." 260(26): 14111-14119;

[0183] (31) P2X5 (Pudding -based receptor P2X Ligandgate Control Ion Channel 5, an ion channel controlled by outside cell ATP, has been involved in synaptic transmission and neurological occurrence, and deficiency can contribute to the pathological physiology of unstable urinary muscles); 422 AA), pi: 7.63, MW:, MW:. 47206 TM: 1 [P] Gene chromosome: 17p13.3, GenBank deposit number np_002552.2) Le et al. (1997) FEBS Lett. 418(1-2):195-199; WO2004047749; WO2003072035 (claim 10); Touchman et al. (2000) Genome Res. 003093444 (claim 1), WO 2003087768 (claim 1); WO 2003029277 (page 82);

[0184] (32) CD72 (B cell differentiation antigen CD72, Lyb-2) PROTEIN SEQUENCE Full maeaity... tafrfpd (1..359; 359aa), pI: 8.66, MW: 40225 TM: 1 [P]Gene Chromosome: 9p13.3, Genbank Accession No. NP_001773.1) WO2004042346 (claim 65); WO2003026493 (51-52, 57-58); WO200075655 (105-106); Von Hoegen et al. (1990) J. Immunol. 144(12):4870-4877; 002) "Proc. Natl. Acad. Sci USA" 99: 16899-16903;

[0185] (33) LY64 (lymphocyte antigen 64 (RP105), a type I membrane protein of the leucine-rich repeat (LRR) family, regulates B-cell activation and apoptosis; loss of function is associated with increased disease activity in patients with systemic lupus erythematosus); .1) U.S. Patent Application Publication No. 2002193567; WO9707198 (claim 11 pp. 39-42); Miura et al. (1996) Genomics 38(3):299-304; Miura et al. (1998) Blood 92:2815-2822; 744452 (claim 8, pages 57-61); International Publication No. 200012130 (pages 24-26);

[0186] (34) FcRH1 (Fc receptor-like protein 1, a putative receptor for immunoglobulin Fc domains containing a C2-type Ig-like domain and an ITAM domain, may have a role in B lymphocyte differentiation); 429 aa, pI: 5.28, MW: 46925 TM: 1 [P] gene chromosome: 1q21-1q22, Genbank Accession No. NP_443170.1) WO 2003077836; WO 200138490 (claim 6, Figures 18E-1 to 18-E-2); Davis et al. (2001) Proc. Natl. Acad. Sci USA 98(17): 9772-9777; Claim 1); WO2003089624 (Claim 7);

[0187] (35) FCRH5 (IRTA2, immunoglobulin superfamily receptor translocation-associated 2, putative immunoreceptor with possible role in B-cell development and lymphopoiesis; translocation-induced gene deregulation occurs in several B-cell malignancies); 977aa, pI: 6.88 MW: 106468 TM: 1 [P] gene chromosome: 1q21, Genbank accession numbers human: AF343662, AF343663, AF3 43664, AF343665, AF369794, AF397453, AK090423, AK090475, AL834187, AY358085; Mouse: AK089756, AY158090, AY506558; NP_112571.1 WO2003024392 (claim 2, Figure 97); Nakayama et al. (2000) "Biochem. Biophys. Res. Commun." 277(1): 124-127; WO2003077836;

[0188] (36) TENB2 (related to TMEFF2, tomoregulin, TPEF, HPP1, TR, putative transmembrane proteoglycan, EGF / hellegren family of growth factors and follistatin); 374 aa, NCBI Accession: AAD55776, AAF91397, AAG49451, NCBI Reference Sequence: NP_057276; NCBI Gene: 23671; OMIM: 6057 34; SwissProt Q9UIK5; Genbank accession number AF179274; AY358907, CAF85723, CQ782436 International Publication No. 2004074320 (SEQ ID NO: 810); Japanese Patent Application Publication No. 2004113151 (SEQ ID NO: 2, 4, 8); International Publication No. 2003042661 (SEQ ID NO: 580); International Publication No. 2003009814 (SEQ ID NO: 411); 268 (page 329); WO200190304 (SEQ ID NO:2706); US2004249130; US2004022727; WO2004063355; US2004197325; U.S. Patent Application Publication No. 2003124579; Horie et al. (2000) Genomics 67:146-152; Uchida et al. (1999) Biochem. Biophys. Res. Commun. 266:593-602; Liang et al. lynne-Jones et al. (2001) "Int J Cancer." Oct 15;94(2):178-84;

[0189] (37) PMEL17 (silver homolog; SILV; D12S53E; PMEL17; SI; SIL); ME20; gp100) BC001414; BT007202; M32295; M77348; NM_006928; McGlinchey, R.P. et al. 106(33), pp. 13731-13736; Kummer, M.P. et al. (2009) J. Biol. Chem. 284(4), pp. 2296-2306;

[0190] (38) TMEFF1 (transmembrane protein 1 with EGF-like domain and two follistatin-like domains; tomoregulin-1); H7365; C9orf2; C9ORF2; U19878; X83961; NM_080655; NM_003692; 629; Gery, S. et al. (2003) Oncogene 22(18):2723-2727;

[0191] (39) GDNF-Ra1 (GDNF family receptor alpha 1; GFRA1; GDNFR; GDNFRA; RETL1; TRNR1; RET1L; GDNFR-alpha1; GFR-ALPHA-1); U95847; BC014962; NM_145793NM_005264; 2264-2277; Treanor, J.J. et al. (1996) Nature 382(6586):80-83;

[0192] (40) Ly6E (lymphocyte antigen-6 complex, locus E; Ly67, RIG-E, SCA-2, TSA-1); NP_002337.1; NM_002346.2; de Nooij-van Dalen, A.G. et al. 2 years) "Mol. Cell. Biol." 22(3): 946-952; International Publication No. 2013 / 17705;

[0193] (41) TMEM46 (shisa homolog 2 (Xenopus laevis); SHISA2); NP_001007539.1; NM_001007538.1; Furushima, K. et al. (2007) Dev. Biol. 306(2), 480-492; Clark, H.F. et al. 3(10): pp. 2265-2270;

[0194] (42) Ly6G6D (lymphocyte antigen-6 complex, locus G6D; Ly6-D, MEGT1); NP_067079.2; NM_021246.2; Mallya, M. et al. (2002) Genomics 80(1):113-123; Ribas, G. et al. (1999) J. Immunol. 163(1):2 pp. 78-287;

[0195] (43) LGR5 (leucine-rich repeat-containing G protein-coupled receptor 5; GPR49, GPR67); NP_003658.1; NM_003667.2; Salanti, G. et al. (2009) Am. J. Epidemiol. 170(5):537-545; Yamamoto, Y. et al. pp. 28-533;

[0196] (44) RET (ret proto-oncogene; MEN2A; HSCR1; MEN2B; MTC1; PTC; CDHF12; Hs.168114; RET51; RET-ELE1); NP_066124.1; NM_020975.4; Tsukamoto, H. et al. (2009) Cancer Sci. arita, N. et al. (2009) Oncogene 28(34): 3058-3068;

[0197] (45) LY6K (lymphocyte antigen-6 complex, locus K; LY6K; HSJ001348; FLJ35226); NP_059997.3; NM_017527.3; Ishikawa, N. et al. (2007) "Cancer Res." 67(24): 11601-11611; 103(6): pp.768-774;

[0198] (46) GPR19 (G protein-coupled receptor 19; Mm.4787); NP_006134.1; NM_006143.2; Montpetit, A. and Sinnett, D. (1999) Hum. Genet. 105(1-2):162-164; O'Dowd, B.F. et al. 94(3): 325-329;

[0199] (47) GPR54 (KISS1 receptor; KISS1R; GPR54; HOT7T175; AXOR12); NP_115940.2; NM_032551.4; Navenot, JM. et al. (2009) Mol. Pharmacol. 29(2):617-623;

[0200] (48) ASPHD1 (Aspartate beta-hydroxylase domain containing 1; LOC253982); NP_859069.2; NM_181718.3; Gerhard, D.S. et al. (2004) Genome Res. 14(10B):2121-2127;

[0201] (49) Tyrosinase (TYR; OCAIA; OCA1A; Tyrosinase; SHEP3); NP_000363.1; NM_000372.4; Bishop, D.T. et al. (2009) Nat. Genet. 41(8):920-925; Nan, H. et al. ~917 pages;

[0202] (50) TMEM118 (ring finger protein, transmembrane 2, RNFT2, FLJ14627); NP_001103373.1; NM_001109903.1; Clark, H.F. et al. (2003) Genome Res. 13(10):2265-2270; Scherer, S.E. et al. 40(7082):pp.346-351

[0203] (51) GPR172A (G protein-coupled receptor 172A; GPCR41; FLJ11856; D15Ertd747e); NP_078807.1; NM_024531.3; Ericsson, T.A. et al. (2003) Proc. Natl. Acad. 4; Takeda, S. et al. (2002) FEBS Lett. 520(1-3):97-101.

[0204] (52) CD33, a member of the immunoglobulin-like lectin family that binds sialic acid, is a 67 kDa glycosylated transmembrane protein. CD33 is expressed on most myeloid and monocytic leukemia cells, in addition to committed myelomonocytic and erythroid progenitors. It is not found in early pluripotent stem cells, mature granulocytes, lymphocytes, or non-hematopoietic cells (Sabbath et al. (1985) J. Clin. Invest. 75:756-56; Andrews et al. (1986) Blood 68:1030-5). CD33 contains two tyrosine residues in its cytoplasmic tail, each of which is followed by hydrophobic residues similar to immunoreceptor tyrosine-based inhibition motifs (ITIMs) found in many inhibitory receptors.

[0205] (53) CLL-1 (CLEC12A, MICL, and DCAL2) encodes a member of the C-type lectin / C-type lectin-like domain (CTL / CTLD) superfamily. Members of this family share a common protein fold and have diverse functions, including cell adhesion, intercellular signaling, glycoprotein turnover, and roles in inflammation and immune responses. The protein encoded by this gene is a negative regulator of granulocyte and monocyte function. Several alternatively spliced ​​transcript variants of this gene have been described, but the full-length nature of some of these variants has not been determined. This gene is closely linked to other CTL / CTLD superfamily members within the natural killer gene complex region on chromosome 12p13 (Drickamer K (1999) Curr. Opin. Struct. Biol. 9(5):585-90; van Rhenen A et al. (2007) Blood 110(7):2659-66; Chen CH et al. 107(4):1459-67; Marshall AS et al. (2006) Eur. J. Immunol. 36(8):2159-69; Bakker AB et al. (2005) Cancer Res. 64(22):8443-50; Marshall AS et al. 92-802). CLL-1 has been shown to be a type II transmembrane receptor containing a single C-type lectin-like domain (not predicted to bind either calcium or sugars), a stalk region, a transmembrane domain, and a short cytoplasmic tail containing the ITIM motif.

[0206] In one aspect, Ab-CIDE antibodies can be against proteins found on many cell or tissue types. Examples of such antibodies include gD and EpCAM. Epithelial cell adhesion molecule (EpCAM) is a transmembrane glycoprotein that mediates Ca2+-independent homotypic cell-cell adhesion in epithelia (Litvinov, S. et al. (1994) Journal of Cell Biology 125(2):437-46). Also known as DIAR5, EGP-2, EGP314, EGP40, ESA, HNPCC8, KS1 / 4, KSA, M4S1, MIC18, MK-1, TACSTD1, TROP1, EpCAM, cell signaling (Maetzel, D. et al. (2009) Nature Cell Biology 11(2):162-71), migration (Osta, WA; et al. (200) 4), "Cancer Res.", 64(16):5818-24), proliferation and differentiation (Litvinov, S. et al. (1996) "Am J Pathol." 148(3):865-75). In addition, EpCAM has oncogenic potential through its ability to upregulate c-myc, e-fabp and cyclins A and E (Munz, M. et al. (2004) Oncogene 23(34):5748-58). Since EpCAM is exclusively expressed in epithelia and epithelial-derived neoplasms, EpCAM can be used as a diagnostic marker for various cancers. In other words, Ab-CIDE can be used to deliver CIDE to many cells or tissues rather than specific cell or tissue types as is the case with targeting antibodies.

[0207] As described herein, Ab-CIDE may comprise an antibody, such as an antibody selected from:

[0208] Anti-Ly6E antibody In certain embodiments, Ab-CIDE can comprise an anti-Ly6E antibody. Lymphocyte antigen-6 complex, locus E (Ly6E), also known as retinoic acid-inducible gene E (RIG-E) and stem cell antigen 2 (SCA-2). Unknown function of a GPI-linked 131 amino acid long, approximately 8.4 kDa protein with an unknown binding partner. It was first identified as a transcript expressed in mouse immature thymocytes, thymic medullary epithelial cells (Mao et al. (1996) Proc. Natl. Acad. Sci. U.S.A. 93:5910-5914). In some embodiments, the subject matter described herein provides Ab-CIDE comprising an anti-Ly6E antibody as described in PCT Publication No. WO2013 / 177055.

[0209] In some embodiments, the subject matter described herein is (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:12, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:13, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:14, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:9, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:10, and (f) HVR-L comprising the amino acid sequence of SEQ ID NO:11. Ab-CIDEs comprising anti-Ly6E antibodies comprising at least 1, 2, 3, 4, 5, or 6 HVRs selected from 3 are provided.

[0210] In one aspect, the subject matter described herein provides an Ab-CIDE comprising an antibody comprising at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:12, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:13, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:14. In further embodiments, the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:12, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:13, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:14.

[0211] In another aspect, the subject matter described herein provides an Ab-CIDE comprising an antibody comprising at least one, at least two, or all three VL HVR sequences selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:9, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:10, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:11. In one embodiment, the antibody comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:9, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:10, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:11.

[0212] In another embodiment, the Ab-CIDE is a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:12, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:13, and (iii) HVR-H3 comprising an amino acid sequence selected from SEQ ID NO:14, and (b) (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO:9, (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO:10, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:11.

[0213] In another aspect, the subject matter described herein includes (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:12, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:13, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:14, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:9, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:10, and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:11. Ab-CIDE is provided, including antibodies comprising:

[0214] In any of the above embodiments, the Ab-CIDE anti-Ly6E antibody is humanized. In one embodiment, the anti-Ly6E antibody comprises an HVR as in any of the above embodiments and further comprises a human acceptor framework, eg a human immunoglobulin framework or a human consensus framework.

[0215] In another embodiment, the Ab-CIDE anti-Ly6E antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:8. In certain embodiments, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:8 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-Ly6E antibodies comprising that sequence retain the ability to bind Ly6E. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:8. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:8. In certain embodiments, substitutions, insertions, or deletions occur within regions outside the HVR (ie, within FRs). Optionally, the anti-Ly6E antibody comprises the VH sequence of SEQ ID NO:8, including post-translational modifications of that sequence. In certain embodiments, the VH comprises two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:12, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:13, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:14.

[0216] In another aspect, an Ab-CIDE anti-Ly6E antibody is provided, the antibody comprising a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:7. In certain embodiments, a VKL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:7 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-Ly6E antibodies containing that sequence retain the ability to bind Ly6E. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:7. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:7. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, within the FRs). Optionally, the anti-Ly6E antibody comprises the VL sequence of SEQ ID NO:7, including post-translational modifications of that sequence. In certain embodiments, the VL comprises one, two, or three HVRs selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:9, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:10, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:11.

[0217] In another aspect there is provided an Ab-CIDE comprising an anti-Ly6E antibody, which antibody comprises a VH as in any of the above embodiments and a VL as in any of the above embodiments.

[0218] In one embodiment, an Ab-CIDE is provided, the antibody comprising the VH and VL sequences of SEQ ID NO:8 and SEQ ID NO:7, respectively, including post-translational modifications of those sequences.

[0219] In a further aspect, provided herein is an Ab-CIDE comprising an antibody that binds to the same epitope as an anti-Ly6E antibody provided herein. For example, in certain embodiments, Ab-CIDE is provided that includes an antibody that binds to the same epitope as an anti-Ly6E antibody that includes the VH sequence of SEQ ID NO:8 and the VL sequence of SEQ ID NO:7, respectively.

[0220] In a further aspect, the Ab-CIDE anti-Ly6E antibody according to any of the above embodiments is a monoclonal antibody, including a human antibody. In one embodiment, the Ab-CIDE anti-Ly6E antibody is an antibody fragment such as Fv, Fab, Fab', scFv, diabodies, or F(ab') 2 Fragments. In another embodiment, the antibody is a substantially full-length antibody, eg, an IgG1 antibody, IgG2a antibody, or other antibody class or isotype as defined herein. In some embodiments, the Ab-CIDE comprises an anti-Ly6E antibody comprising heavy and light chains comprising the amino acid sequences of SEQ ID NOs: 16 and 15, respectively

[0221] Anti-HER2 antibody In certain embodiments, Ab-CIDE comprises an anti-HER2 antibody. In one embodiment, the Ab-CIDE anti-HER2 antibodies comprise humanized anti-HER2 antibodies, such as huMAb4D5-1, huMAb4D5-2, huMAb4D5-3, huMAb4D5-4, huMAb4D5-5, huMAb4D5-6, huMAb4D5-7 and huMAb4D5-8, as described in Table 3 of U.S. Patent No. 5,821,337. These antibodies contain human framework regions with the complementarity determining regions of the murine antibody (4D5) that binds HER2. The humanized antibody huMAb4D5-8 is also called trastuzumab and is commercially available under the trade name HERCEPTIN®. In another embodiment, the Ab-CIDE anti-HER2 antibody comprises a humanized anti-HER2 antibody, such as humanized 2C4, as described in US Pat. No. 7,862,817. An exemplary humanized 2C4 antibody is Pertuzumab, marketed under the trade name PERJETA®.

[0222] In another embodiment, the Ab-CIDE anti-HER2 antibody comprises a humanized 7C2 anti-HER2 antibody. A humanized 7C2 antibody is an anti-HER2 antibody.

[0223] (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:23, 27, or 28; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:24 or 29; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:19; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:20; Ab-CIDEs comprising anti-HER2 antibodies comprising at least 1, 2, 3, 4, 5, or 6 HVRs selected from HVR-L3 comprising sequences are described. (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:23; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:24; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:19; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:20; A PAC comprising an anti-HER2 antibody comprising at least 1, 2, 3, 4, 5, or 6 HVRs is described.

[0224] In one aspect, described herein is an Ab-CIDE comprising an antibody comprising at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:22, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:23, 27, or 28, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:24 or 29. In one aspect, described herein is an Ab-CIDE comprising an antibody comprising at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:22, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:23, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:24. In further embodiments, the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:68, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:23, 27, or 28, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:24 or 29. In further embodiments, the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:22, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:23, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:24.

[0225] In another aspect, described herein is an Ab-CIDE comprising an antibody comprising at least one, at least two, or all three VL HVR sequences selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:19, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:20, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:21. In one embodiment, the antibody comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:19, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:20, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:21.

[0226] In another embodiment, the Ab-CIDE is a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:22, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:23, 27, or 28, and (iii) HVR-H3 comprising the amino acid sequence selected from SEQ ID NO:24 or 29, and (b) (i) the amino acid sequence of SEQ ID NO:19 (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO:20; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:21. In another embodiment, the Ab-CIDE is a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:22, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:23, and (iii) HVR-H3 comprising the amino acid sequence selected from SEQ ID NO:24, and (b) (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO:19, (ii) a) HVR-L2 comprising the amino acid sequence of SEQ ID NO:20; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:21.

[0227] (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:23, 27, or 28; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:24 or 29; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:19; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:20; Ab-CIDEs are described, including antibodies that contain HVR-L3. In another aspect, provided herein is an antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:22, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:23, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:24, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:19, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:20, and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:21. Ab-CIDE is described, including

[0228] In any of the above embodiments, the Ab-CIDE anti-HER2 antibody is humanized. In one embodiment, the Ab-CIDE anti-HER2 antibody comprises an HVR as in any of the above embodiments and further comprises a human acceptor framework, eg, a human immunoglobulin framework or a human consensus framework.

[0229] In another embodiment, the Ab-CIDE anti-HER2 antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:18. In certain embodiments, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 18 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-HER2 antibodies comprising that sequence retain the ability to bind HER2. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:18. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:18. In certain embodiments, substitutions, insertions, or deletions occur within regions outside the HVR (ie, within FRs). Optionally, the anti-HER2 antibody comprises the VH sequence of SEQ ID NO: 18, including post-translational modifications of that sequence. In certain embodiments, the VH comprises two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:22, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:23, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:24.

[0230] In another aspect, an Ab-CIDE anti-HER2 antibody is provided, the antibody comprising a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:17. In certain embodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 17 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-HER2 antibodies comprising that sequence retain the ability to bind HER2. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:17. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:17. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, within the FRs). Optionally, the anti-HER2 antibody comprises the VL sequence of SEQ ID NO: 17, including post-translational modifications of that sequence. In certain embodiments, the VL comprises one, two, or three HVRs selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:19, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:20, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:21.

[0231] In another aspect there is provided an Ab-CIDE comprising an anti-HER2 antibody, the antibody comprising a VH as in any of the above embodiments and a VL as in any of the above embodiments.

[0232] In one embodiment, an Ab-CIDE comprising an antibody is provided, the antibody comprising the VH and VL sequences of SEQ ID NO: 18 and SEQ ID NO: 17, respectively, including post-translational modifications of those sequences.

[0233] In one embodiment, an Ab-CIDE comprising an antibody comprising the humanized 7C2.v2.2.LA(hu7C2)K149C kappa light chain sequence of SEQ ID NO:30 is provided.

[0234] In one embodiment, an Ab-CIDE comprising an antibody comprising the Hu7C2 A118C IgG1 heavy chain sequence of SEQ ID NO:31 is provided.

[0235] In a further aspect, provided herein is a PAC comprising an antibody that binds to the same epitope as an anti-HER2 antibody provided herein. For example, in certain embodiments, Ab-CIDE is provided comprising an antibody that binds to the same epitope as an anti-HER2 antibody comprising the VH sequence of SEQ ID NO:18 and the VL sequence of SEQ ID NO:17, respectively.

[0236] In a further aspect, the Ab-CIDE anti-HER2 antibody according to any of the above embodiments is a monoclonal antibody, including a human antibody. In one embodiment, the Ab-CIDE anti-HER2 antibody is an antibody fragment, e.g., Fv, Fab, Fab', scFv, diabodies, or F(ab') 2 Fragments. In another embodiment, Ab-CIDE comprises a substantially full-length antibody, eg, an IgG1 antibody, an IgG2a antibody, or an antibody of another antibody class or isotype as defined herein.

[0237] Anti-B7-H4 antibody In certain embodiments, Ab-CIDE can comprise an anti-B7-H4 antibody. B7-H4 is a type I transmembrane protein and a member of the B7 superfamily of proteins that provide co-signals in conjunction with T-cell receptor antigenic signals. B7-H4 is a negative regulator of T cell function and ligation of T cells inhibits their growth, cytokine secretion and cytotoxicity. Elimination of B7-H4 in mice does not affect immune cell homeostasis and there are no signs of autoimmunity. Zhu et al. Blood 113(8):1759-1767 (2009); Suh et al. Molecular and Cellular Biology 26(17):6403-6411 (2006).The receptor for B7-H4 is unknown and has not been identified.

[0238] Human B7-H4 is a 282 amino acid protein (including the amino terminal signal sequence), of which approximately 227 amino acids are predicted to be in the extracellular space after cleavage of the amino terminal signal sequence. B7-H4 contains an Ig-like V domain, an Ig-like C domain, a transmembrane domain and a short cytoplasmic tail. B7-H4 is a member of the B7 family that has the potential to downregulate the immune system through its co-inhibitory signals in conjunction with antigen-dependent signaling by T cell receptors. B7-H4 is nominally expressed in normal human tissues, but is highly overexpressed in a myriad of human cancers, including cancers of the female reproductive system (breast, ovary and endometrium). The prevalence of B7-H4 has been reported to be high in invasive ductal and lobular carcinomas, including both primary (-95%) and metastatic (-97%) breast cancer. Increased B7-H4 staining was associated with negative PR and HER2 status, but expression was independent of tumor grade or stage. In addition to the high proportion of B7H4-stained cells in these types of breast cancer, the number of infiltrating lymphocytes was also reduced at the same time. Recently, in a B7-H4 knockout model of lung metastatic breast cancer, the authors reported that B7-H4− / − mice had fewer lung tumor nodules and exhibited improved survival and memory responses to tumor challenge compared with wild-type mice. This was attributed to immunosuppressive effects on CD4 and CD8 cells by tumor-associated neutrophils bound to the B7-H4-Ig fusion protein. This may also explain why transplanted SKOV3 cells overexpressing B7-H4 proliferated more aggressively than wild-type SKOV3 cells in SCID mice. Furthermore, it was shown that B7-H4 mRNA and protein knockdown in SKBR3 cells resulted in increased caspase activity and apoptosis. In some embodiments, the subject matter described herein provides Ab-CIDE comprising an anti-B7-H4 antibody as described in PCT Publication No. WO2016 / 040724.

[0239] In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises: (a) (i) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 128, (ii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 129, and (iii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 200; or (b) (i) HVR-H3 comprising the amino acid sequence of SEQ ID NO:201, (ii) HVR-L3 comprising the amino acid sequence of SEQ ID NO:129, and (iii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:200.

[0240] In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises: (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 199, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 200, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 128; or (b) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:199, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:200, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO:201.

[0241] In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises the heavy chain framework FR3 sequence of SEQ ID NO:213.

[0242] In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises: (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:202, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:129. In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises the light chain framework FR3 sequence of SEQ ID NO:207.

[0243] In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises: (a) a VH sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 198; (b) a VL sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 126; or (c) a VH sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 127; or (d) a VH sequence as in (a) and a VL sequence as in (b); or (e) VH sequences as in (c) and VL sequences as in (b).

[0244] In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises the VH sequence of SEQ ID NO:198 or 127. In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises the VL sequence of SEQ ID NO:126.

[0245] In some embodiments, an Ab-CIDE anti-B7-H4 antibody comprising (a) the VH sequence of SEQ ID NO: 198 and the VL sequence of SEQ ID NO: 126, or (b) the VH sequence of SEQ ID NO: 127 and the VL sequence of SEQ ID NO: 126.

[0246] In some embodiments, Ab-CIDE anti-B7-H4 antibodies are provided, the antibodies comprising: (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:199, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:200, (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO:128, (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO:202, (v) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203, and (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO:129; (b) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:199, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:200, (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO:201, (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO:202, (v) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203, and (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO:129.

[0247] In any of the embodiments described herein, the Ab-CIDE anti-B7-H4 antibody can be a monoclonal antibody. In any of the embodiments described herein, the Ab-CIDE anti-B7-H4 antibody can be a human, humanized, or chimeric antibody. In any of the embodiments described herein, the Ab-CIDE anti-B7-H4 antibody can be an antibody fragment that binds B7-H4. Antibody fragments include Fab, Fab', Fab'-SH, F(ab') 2 , Fv, and scFv fragments, as well as other fragments described below.

[0248] In any of the embodiments described herein, the Ab-CIDE anti-B7-H4 antibody can be an IgG1, IgG2a or IgG2b antibody. In any of the embodiments described herein, the Ab-CIDE anti-B7-H4 antibody may comprise one or more engineered cysteine ​​amino acid residues. In any of the embodiments described herein, one or more engineered cysteine ​​amino acid residues may be located in the heavy chain. In any of the embodiments described herein, one or more engineered cysteine ​​amino acid residues may be located in the light chain. In any of the embodiments described herein, the Ab-CIDE anti-B7-H4 antibody may comprise at least one mutation in the heavy chain constant region selected from A118C and S400C. In any of the embodiments described herein, the Ab-CIDE anti-B7-H4 antibody may comprise at least one mutation in the light chain constant region selected from K149C and V205C.

[0249] In some embodiments, an Ab-CIDE anti-B7-H4 is provided, wherein the antibody has (a) the heavy chain sequence of SEQ ID NO: 132 and the light chain sequence of SEQ ID NO: 134; or (b) the heavy chain sequence of SEQ ID NO: 133 and the light chain sequence of SEQ ID NO: 134; or (c) the heavy chain sequence of SEQ ID NO: 130 and the light chain sequence of SEQ ID NO: 140; (e) the heavy chain sequence of SEQ ID NO: 131 and the light chain sequence of SEQ ID NO: 140; or (f) the heavy chain sequence of SEQ ID NO: 131 and the light chain sequence of SEQ ID NO: 141; or (g) the heavy chain sequence of SEQ ID NO: 144 and the light chain sequence of SEQ ID NO: 142; or (d) the heavy chain sequence of SEQ ID NO: 130 and the light chain sequence of SEQ ID NO: 146; or (e) the heavy chain sequence of SEQ ID NO: 131 and the light chain sequence of SEQ ID NO: 145; or (f) the heavy chain sequence of SEQ ID NO: 131 and the light chain sequence of SEQ ID NO: 146; and the light chain sequence of SEQ ID NO:148.

[0250] In some embodiments, the Ab-CIDE anti-B7-H4 antibody is a bi-epitopic antibody comprising a first half-antibody and a second half-antibody provided that the first half-antibody comprises a first VH / VL unit that binds a first epitope of B7-H4 and the second half-antibody comprises a second VH / VL unit that binds a second epitope of B7-H4. In some embodiments, the first epitope or the second epitope is an epitope within all or part of the B7-H4Ig-V containing domain. In some embodiments, the first epitope or the second epitope is not within the B7-H4Ig-V domain or entirely not within the B7-H4Ig-V containing domain. In some embodiments, the first epitope is not within all or part of the B7-H4Ig-V containing domain and the second epitope is not within the B7-H4Ig-V domain or not completely within the B7-H4Ig-V containing domain; within all or part of the B7-H4Ig-V containing domain. In some embodiments, the first epitope and the second epitope are each independently selected from: a) an epitope within all or part of the B7-H4Ig-V containing domain; b) an epitope within all or part of the B7-H4Ig-C-containing domain; and c) Epitopes within all or part of the B7-H4Ig-V and Ig-C containing domains.

[0251] In some embodiments, the B7-H4Ig-V containing domain has the sequence of amino acids 29-157 of SEQ ID NO:233. In some embodiments, the B7-H4Ig-C-containing domain has the sequence of amino acids 158-250 of SEQ ID NO:233.

[0252] In some embodiments, a) a first half-antibody binds an epitope within all or part of the B7-H4Ig-V-containing domain and a second half-antibody binds an epitope within all or part of the B7-H4Ig-C-containing domain; or b) the first half-antibody binds an epitope within all or part of the B7-H4Ig-V containing domain and the second half-antibody binds an epitope within all or part of the B7-H4Ig-V and Ig-C containing domain; or c) the first half-antibody binds an epitope within all or part of the B7-H4Ig-C-containing domain and the second half-antibody binds an epitope within all or part of the B7-H4Ig-V and Ig-C-containing domain; or d) the first half-antibody binds an epitope within all or part of the B7-H4Ig-C-containing domain and the second half-antibody binds an epitope within all or part of the B7-H4Ig-V-containing domain; or e) the first half-antibody binds an epitope within all or part of the B7-H4Ig-V and Ig-C-containing domain and the second half-antibody binds an epitope within all or part of the B7-H4Ig-V-containing domain; or f) the first half-antibody binds an epitope within all or part of the B7-H4Ig-V and Ig-C-containing domain and the second half-antibody binds an epitope within all or part of the B7-H4Ig-C-containing domain.

[0253] In some embodiments, the first half-antibody binds an epitope within all or part of the B7-H4Ig-V-containing domain and the second half-antibody binds an epitope within all or part of the B7-H4Ig-V and Ig-C-containing domain; Binds an epitope within all or part of the H4Ig-V containing domain.

[0254] In some embodiments, the first half-antibody comprises: (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 199, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 200, (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 128, (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 202, (v) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 203, and (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 129; (b) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:199, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:200, (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO:201, (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO:202, (v) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203, and (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO:129; (c) the VH sequence of SEQ ID NO: 198 and the VL sequence of SEQ ID NO: 126; or (d) VH sequence of SEQ ID NO:127 and VL sequence of SEQ ID NO:126.

[0255] In some embodiments, the second half-antibody comprises: (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 199, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 200, (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 128, (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 202, (v) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 203, and (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 129; (b) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:199, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:200, (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO:201, (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO:202, (v) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203, and (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO:129; (c) the VH sequence of SEQ ID NO: 198 and the VL sequence of SEQ ID NO: 126; or (d) VH sequence of SEQ ID NO:127 and VL sequence of SEQ ID NO:126.

[0256] In some embodiments, the first half-antibody comprises: (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:218, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:219, (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO:220, (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO:221, (v) HVR-L2 comprising the amino acid sequence of SEQ ID NO:222, and (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO:223; or (b) VH sequence of SEQ ID NO:216 and VL sequence of SEQ ID NO:215.

[0257] In some embodiments, the second half-antibody comprises: (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:218, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:219, (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO:220, (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO:221, (v) HVR-L2 comprising the amino acid sequence of SEQ ID NO:222, and (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO:223; or (b) VH sequence of SEQ ID NO:216 and VL sequence of SEQ ID NO:215.

[0258] In some embodiments, the Ab-CIDE anti-B7-H4 antibody is a bi-epitopic antibody that is an IgG1 or IgG4 antibody. In some embodiments, the first half-antibody comprises a first heavy chain constant region comprising a knob mutation and the second heavy chain comprises a second heavy chain constant region comprising a hole mutation; or the first half-antibody comprises a first heavy chain constant region comprising a hole mutation and the second heavy chain comprises a second heavy chain constant region comprising a knob mutation. In some embodiments, the bi-epitopic antibody is an IgG1 antibody and the knob mutation comprises the T366W mutation. In some embodiments, the bi-epitopic antibody is an IgG1 antibody and the Hall mutation comprises at least one, at least two or three mutations selected from T366S, L368A and Y407V. In some embodiments, the bi-epitopic antibody is an IgG4 antibody and the knob mutation comprises the T366W mutation. In some embodiments, the bi-epitopic antibody is an IgG4 antibody and the Hall mutation comprises at least one, at least two, or three mutations selected from T366S, L368A and Y407V mutations.

[0259] In some embodiments, an Ab-CIDE anti-B7-H4 antibody that is a bi-epitopic antibody is provided: a) the first half-antibody comprises the heavy chain sequence of SEQ ID NO: 159 or 163 and the light chain sequence of SEQ ID NO: 145 or 146; b) the first half-antibody comprises the heavy chain sequence of SEQ ID NO: 160 or 164 and the light chain sequence of SEQ ID NO: 145 or 146; c) the first half-antibody comprises the heavy chain sequence of SEQ ID NO: 161 or 165 and the light chain sequence of SEQ ID NO: 147 or 148; d) the first half-antibody comprises the heavy chain sequence of SEQ ID NO: 162 or 166 and the light chain sequence of SEQ ID NO: 147 or 148; e) the second half-antibody comprises the heavy chain sequence of SEQ ID NO: 159 or 163 and the light chain sequence of SEQ ID NO: 145 or 146; f) the second half-antibody comprises the heavy chain sequence of SEQ ID NO: 160 or 164 and the light chain sequence of SEQ ID NO: 145 or 146; g) the second half-antibody comprises the heavy chain sequence of SEQ ID NO: 161 or 165 and the light chain sequence of SEQ ID NO: 147 or 148; or h) the second half-antibody comprises the heavy chain sequence of SEQ ID NO:162 or 166 and the light chain sequence of SEQ ID NO:147 or 148;

[0260] In some embodiments, an Ab-CIDE anti-B7-H4 antibody that is a bi-epitopic antibody is provided: or b) the first half antibody comprises the heavy chain sequence of SEQ ID NO: 161 or 165 and the light chain sequence of SEQ ID NO: 147 or 148 and the second half antibody comprises the heavy chain sequence of SEQ ID NO: 160 or 164 and the light chain sequence of SEQ ID NO: 145 or 146.

[0261] In some embodiments, an Ab-CIDE anti-B7-H4 antibody that is a bi-epitopic antibody, comprising a first half-antibody and a second half-antibody, the first half-antibody comprising a first VH / VL unit that binds a first epitope of B7-H4, the second half-antibody comprising a second VH / VL unit that binds a second epitope of B7-H4, the first half-antibody comprising SEQ ID NO: 159 Or provided is an anti-B7-H4 antibody comprising the heavy chain sequence of 163 and the light chain sequence of SEQ ID NO:145, wherein the second half antibody comprises the heavy chain sequence of SEQ ID NO:162 or 166 and the light chain sequence of SEQ ID NO:147.

[0262] In any of the embodiments described herein, B7-H4 can be human B7-H4 of SEQ ID NO:233.

[0263] An exemplary native human B7-H4 precursor protein sequence with signal sequence (amino acids 1-28) is provided in SEQ ID NO:233 and the corresponding mature B7-H4 protein sequence is shown in SEQ ID NO:234 ​​(corresponding to amino acids 29-282 of SEQ ID NO:233).

[0264] In certain embodiments, an anti-B7-H4 antibody has one or more of the following characteristics, in any combination: (a) binds to an epitope within all or part of the B7-H4Ig-V-containing domain (amino acids 29-157 of SEQ ID NO:233); or binds to an epitope within all or part of the B7-H4Ig-C-containing domain (amino acids 158-250 of SEQ ID NO:233); 250); or binds to an epitope within all or part of SEQ ID NO:234 ​​(mature human B7-H4); or binds to an epitope within all or part of SEQ ID NO:233 (human B7-H4 precursor); (b) binds B7-H4 with an affinity of ≦100 nM, ≦50 nM, ≦10 nM, or ≦9 nM, or ≦8 nM, or ≦7 nM, or ≦6 nM, or ≦5 nM, or ≦4 nM, or ≦3 nM, or ≦2 nM, or ≦1 nM, optionally ≧0.0001 nM, or ≧0.001 nM, or ≧0.01 nM.

[0265] Non-limiting exemplary anti-B7-H4 antibodies of Ab-CIDE include hu1D11.v1.9 varC2 and hu1D11.v1.9 varD described herein. In some embodiments, B7-H4 is human B7-H4. In some embodiments, B7-H4 is selected from human, cynomolgus monkey, mouse, and rat B7-H4.

[0266] In some embodiments, the Ab-CIDE anti-B7-H4 antibody binds to an epitope within all or part of the B7-H4Ig-V-containing domain (amino acids 29-157 of SEQ ID NO:233). In some embodiments, the Ab-CIDE anti-B7-H4 antibody binds to an epitope within all or part of the B7-H4Ig-C-containing domain (amino acids 158-250 of SEQ ID NO:233). In some embodiments, the Ab-CIDE anti-B7-H4 antibody binds to an epitope within all or part of the B7-H4Ig-V and Ig-C containing domains (amino acids 29-250 of SEQ ID NO:233). In some embodiments, the Ab-CIDE anti-B7-H4 antibody binds to an epitope within all or part of SEQ ID NO: 234 (mature human B7-H4). In some embodiments, the Ab-CIDE anti-B7-H4 antibody binds to an epitope within all or part of SEQ ID NO: 233 (human B7-H4 precursor). In some such embodiments, the Ab-CIDE anti-B7-H4 antibody is ≦100 nM, ≦50 nM, ≦10 nM, or ≦9 nM, or ≦8 nM, or ≦7 nM, or ≦6 nM, or ≦5 nM, or ≦4 nM, or ≦3 nM, or ≦2 nM, or ≦1 nM, optionally ≧0.0001 nM, or ≧0.001 nM, or Binds B7-H4 with an affinity ≧0.01 nM.

[0267] Antibody 1D11v1.9 variants and other embodiments In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:199, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:200, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:128, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:202, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203, and (f) the sequence comprising at least 1, 2, 3, 4, 5, or 6 HVRs selected from HVR-L3 comprising the amino acid sequence numbered 129;

[0268] In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:199, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:200, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:201, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:202, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203, and (f) the sequence comprising at least 1, 2, 3, 4, 5, or 6 HVRs selected from HVR-L3 comprising the amino acid sequence numbered 129;

[0269] In one aspect, the Ab-CIDE anti-B7-H4 antibody comprises at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 199, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 200, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 128. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:201. In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:128 and HVR-L3 comprising the amino acid sequence of SEQ ID NO:129. In further embodiments, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:128, HVR-L3 comprising the amino acid sequence of SEQ ID NO:129, and HVR-H2 comprising the amino acid sequence of SEQ ID NO:200. In a further embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:199, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:200, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:128.

[0270] In one aspect, the Ab-CIDE anti-B7-H4 antibody comprises at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 199, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 200, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 201. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:201. In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:201 and HVR-L3 comprising the amino acid sequence of SEQ ID NO:129. In further embodiments, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:201, HVR-L3 comprising the amino acid sequence of SEQ ID NO:129, and HVR-H2 comprising the amino acid sequence of SEQ ID NO:200. In a further embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 199, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 200, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 201.

[0271] In another aspect, Ab-CIDE anti-B7-H4 antibodies are provided that comprise at least one, at least two, or all three VL HVR sequences selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:202, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:129. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:202, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:129.

[0272] In another embodiment, the antibody B7-H4 antibody of Ab-CIDE is a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 199, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 200, and (iii) HVR-H3 comprising the amino acid sequence selected from SEQ ID NO: 128, and (b) (i) SEQ ID NO: 202 (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:129.

[0273] In another embodiment, the antibody B7-H4 antibody of Ab-CIDE is a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 199, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 200, and (iii) HVR-H3 comprising the amino acid sequence selected from SEQ ID NO: 201, and (b) (i) SEQ ID NO: 202 (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:129.

[0274] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:199; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:200; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:128; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:202; Includes HVR-L3 containing 29 amino acid sequences.

[0275] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:199; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:200; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:201; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:202; Includes HVR-L3 containing 29 amino acid sequences.

[0276] In any of the above embodiments, the Ab-CIDE anti-B7-H4 antibody is humanized. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises an HVR as in any of the above embodiments and further comprises a human acceptor framework, eg, a human immunoglobulin framework or a human consensus framework. In certain embodiments, the human acceptor framework is the human VL kappa I consensus (VL KI ) framework and / or VH framework VH 1 is. In certain embodiments, the human acceptor framework is a human VL kappa I consensus (VL KI ) framework and / or VH framework VH 1 are: Y49H, V58I, T69R and / or F71Y mutations in the light chain framework region FR3; V67A, I69L, R71A, T73K and / or T75S mutations in the heavy chain framework region FR3.

[0277] In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises the HVR of any of the above embodiments and further comprises the heavy chain framework FR3 sequence of SEQ ID NO:213. In some such embodiments, the heavy chain variable domain framework is a modified human VH having the FR3 sequence of SEQ ID NO:213. 1 Framework.

[0278] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 198 or 127. In certain embodiments, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 198 or 127 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-B7-H4 antibodies comprising that sequence retain the ability to bind B7-H4. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted and / or deleted in SEQ ID NO:198 or 127. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:198 or 127. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, in the FRs).

[0279] Optionally, the Ab-CIDE anti-B7-H4 antibody comprises the VH sequence of SEQ ID NO: 198, including post-translational modifications of that sequence. In certain embodiments, the VH comprises two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:199, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:200, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:201. Optionally, the Ab-CIDE anti-B7-H4 antibody comprises the VH sequence of SEQ ID NO: 127, including post-translational modifications of that sequence. In certain embodiments, the VH comprises two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:199, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:200, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:128. In another aspect, an anti-B7-H4 antibody is provided, the antibody comprising a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:126. In certain embodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 126 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-B7-H4 antibodies comprising that sequence retain the ability to bind B7-H4. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:126. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:126. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, in the FRs). Optionally, the Ab-CIDE anti-B7-H4 antibody comprises the VL sequence of SEQ ID NO: 126, including post-translational modifications of that sequence. In certain embodiments, the VL comprises one, two, or three HVRs selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:202, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:129.

[0280] In another aspect, Ab-CIDE anti-B7-H4 antibodies are provided, wherein the antibody comprises a VH as in any of the embodiments provided above and a VL as in any of the embodiments provided above.

[0281] In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 198 and SEQ ID NO: 126, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 127 and SEQ ID NO: 126, respectively, including post-translational modifications of those sequences.

[0282] In certain embodiments, provided is an Ab-CIDE anti-B7-H4 antibody according to any of the above embodiments that binds to B7-H4 and has at least one of the following characteristics: (a) binds to an epitope within all or part of the B7-H4Ig-V-containing domain (amino acids 29-157 of SEQ ID NO:233); or an epitope within all or part of the B7-H4 Ig-V and Ig-C domains (amino acids 29-250 of SEQ ID NO:233); or an epitope within all or part of SEQ ID NO:234 ​​(mature human B7-H4); or an epitope within all or part of SEQ ID NO:233 (human B7-H4 precursor). In some embodiments, the anti-B7-H4 antibody has one or more of the following characteristics, in any combination: (a) binds to an epitope within all or part of the B7-H4Ig-V-containing domain (amino acids 29-157 of SEQ ID NO:233); or binds to an epitope within all or part of the B7-H4Ig-C-containing domain (amino acids 158-250 of SEQ ID NO:233); Binds epitopes within all or part of the g-V and Ig-C domains (amino acids 29-250 of SEQ ID NO:233); or binds epitopes within all or part of SEQ ID NO:234 ​​(mature human B7-H4); or binds epitopes within all or part of SEQ ID NO:233 (human B7-H4 precursor).

[0283] In a further aspect, the Ab-CIDE anti-B7-H4 antibody according to any of the above embodiments is a monoclonal antibody, including chimeric, humanized, or human antibodies. In one embodiment, the Ab-CIDE anti-B7-H4 antibody is an antibody fragment, e.g., Fv, Fab, Fab', scFv, diabodies, or F(ab') 2 Fragments. In another embodiment, the Ab-CIDE anti-B7-H4 antibody is a substantially full-length antibody, eg, an IgG1 antibody as defined herein, or other antibody class or isotype.

[0284] Antibody 1D11 and other embodiments In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:5, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:6, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:167, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:168, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:169, and (f) SEQ ID NO:10. comprising at least 1, 2, 3, 4, 5, or 6 HVRs selected from HVR-L3 comprising amino acid sequences. In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 199, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 200, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 201, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 202, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 203, and (f) SEQ ID NO: comprising at least 1, 2, 3, 4, 5, or 6 HVRs selected from HVR-L3, which comprises a 204 amino acid sequence.

[0285] In one aspect, the Ab-CIDE anti-B7-H4 antibody comprises at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:5, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:6, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:167. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:167. In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:167 and HVR-L3 comprising the amino acid sequence of SEQ ID NO:170. In a further embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:167, HVR-L3 comprising the amino acid sequence of SEQ ID NO:170, and HVR-H2 comprising the amino acid sequence of SEQ ID NO:6. In a further embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:5, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:6, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:167.

[0286] In one aspect, the Ab-CIDE anti-B7-H4 antibody comprises at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 199, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 200, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 201. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:201. In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:201 and HVR-L3 comprising the amino acid sequence of SEQ ID NO:204. In further embodiments, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:201, HVR-L3 comprising the amino acid sequence of SEQ ID NO:204, and HVR-H2 comprising the amino acid sequence of SEQ ID NO:200. In a further embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 199, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 200, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 201.

[0287] In another aspect, Ab-CIDE anti-B7-H4 antibodies are provided that comprise at least one, at least two, or all three VL HVR sequences selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 168, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 169, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 170. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 168, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 169, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 170.

[0288] In another aspect, Ab-CIDE anti-B7-H4 antibodies are provided that comprise at least one, at least two, or all three VL HVR sequences selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:202, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:204. In one embodiment, the antibody comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:202, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:204.

[0289] In another embodiment, the antibody B7-H4 antibody of Ab-CIDE comprises a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:5, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:6, and (iii) HVR-H3 comprising the amino acid sequence selected from SEQ ID NO:167, and (b) (i) the amino acid sequence of SEQ ID NO:168 (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 169; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 170.

[0290] In another embodiment, the antibody B7-H4 antibody of Ab-CIDE is a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 199, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 200, and (iii) HVR-H3 comprising the amino acid sequence selected from SEQ ID NO: 201, and (b) (i) SEQ ID NO: 202 (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:204.

[0291] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:5; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:6; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:167; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:168; Includes HVR-L3 containing sequences.

[0292] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 199, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 200, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 201, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 202, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 203, and (f) SEQ ID NO: Includes HVR-L3 containing 204 amino acid sequences.

[0293] In any of the above embodiments, the Ab-CIDE anti-B7-H4 antibody is humanized. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises an HVR as in any of the above embodiments and further comprises a human acceptor framework, eg, a human immunoglobulin framework or a human consensus framework. In certain embodiments, the human acceptor framework is the human VL kappa I consensus (VL KI ) framework and / or VH framework VH 1 is. In certain embodiments, the human acceptor framework is a human VL kappa I consensus (VL KI ) framework and / or VH framework VH 1 are: Y49H, V58I, T69R and / or F71Y mutations in the light chain framework region FR3; V67A, I69L, R71A, T73K and / or T75S mutations in the heavy chain framework region FR3.

[0294] In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises the HVR of any of the above embodiments and further comprises the heavy chain framework FR3 sequence of SEQ ID NOs:211, 212, or 213. In some such embodiments, the heavy chain variable domain framework is a modified human VH having the FR3 sequence of SEQ ID NO:211, 212, or 213. 1 Framework.

[0295] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:4. In certain embodiments, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:4 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-B7-H4 antibodies comprising that sequence retain the ability to bind B7-H4. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:4. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:4. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, in the FRs). Optionally, the Ab-CIDE anti-B7-H4 antibody comprises the VH sequence of SEQ ID NO: 4, including post-translational modifications of that sequence. In certain embodiments, the VH comprises two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:5, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:6, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:167.

[0296] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 196, 197, 198, 99, 100, 101, 102, or 103. In certain embodiments, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 196, 197, 198, 99, 100, 101, 102, or 103 contains substitutions (e.g., conservative substitutions), insertions, or deletions compared to a reference sequence, but the sequence is Anti-B7-H4 antibodies comprising retain the ability to bind to B7-H4. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 196, 197, 198, 99, 100, 101, 102, or 103. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO: 196, 197, 198, 99, 100, 101, 102, or 103. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, in the FRs). Optionally, the Ab-CIDE anti-B7-H4 antibody comprises the VH sequence of SEQ ID NO: 196, 197, 198, 99, 100, 101, 102, or 103, including post-translational modifications of that sequence. In certain embodiments, the VH comprises two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:199, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:200, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:201.

[0297] In another aspect, an Ab-CIDE anti-B7-H4 antibody is provided, wherein the antibody comprises a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:3. In certain embodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:3 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-B7-H4 antibodies comprising that sequence retain the ability to bind B7-H4. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:3. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:3. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, in the FRs). Optionally, the Ab-CIDE anti-B7-H4 antibody comprises the VL sequence of SEQ ID NO:3, including post-translational modifications of that sequence. In certain embodiments, the VL comprises 1, 2, or 3 HVRs selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 168, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 169, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 170.

[0298] In another aspect, anti-B7-H4 antibodies are provided, said antibodies comprising a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NOs: 195, 253, 254, 255, 256, 257, or 258. In certain embodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 195, 253, 254, 255, 256, 257, or 258 contains a substitution (e.g., conservative substitution), insertion, or deletion compared to the reference sequence, but the anti-B7 -H4 antibodies retain the ability to bind to B7-H4. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NOs:195, 253, 254, 255, 256, 257, or 258. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NOs:195, 253, 254, 255, 256, 257, or 258. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, in the FRs). Optionally, the Ab-CIDE anti-B7-H4 antibody comprises the VL sequence of SEQ ID NO: 195, 253, 254, 255, 256, 257, or 258, including post-translational modifications of that sequence. In certain embodiments, the VL comprises one, two, or three HVRs selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:202, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:203, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:204.

[0299] In another aspect, Ab-CIDE anti-B7-H4 antibodies are provided, wherein the antibody comprises a VH as in any of the embodiments provided above and a VL as in any of the embodiments provided above.

[0300] In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO:4 and SEQ ID NO:3, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 101 and SEQ ID NO: 253, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 101 and SEQ ID NO: 257, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 102 and SEQ ID NO: 258, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 103 and SEQ ID NO: 258, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 101 and SEQ ID NO: 256, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 101 and SEQ ID NO: 255, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 101 and SEQ ID NO: 254, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 100 and SEQ ID NO: 253, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO:99 and SEQ ID NO:253, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 196 and SEQ ID NO: 253, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 196 and SEQ ID NO: 195, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 197 and SEQ ID NO: 195, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 198 and SEQ ID NO: 195, respectively, including post-translational modifications of those sequences.

[0301] In a further aspect, provided herein is an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope as the anti-B7-H4 antibody. For example, in certain embodiments, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:4 and the VL sequence of SEQ ID NO:3 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope. In a specific embodiment, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:101 and the VL sequence of SEQ ID NO:253 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope. In a specific embodiment, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:101 and the VL sequence of SEQ ID NO:257 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope. In a specific embodiment, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:102 and the VL sequence of SEQ ID NO:258 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope. In a specific embodiment, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:103 and the VL sequence of SEQ ID NO:258 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope. In a specific embodiment, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:101 and the VL sequence of SEQ ID NO:256 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope. In a specific embodiment, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:101 and the VL sequence of SEQ ID NO:255 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope. In a specific embodiment, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:101 and the VL sequence of SEQ ID NO:254 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope. In a specific embodiment, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:100 and the VL sequence of SEQ ID NO:253 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope. In a specific embodiment, an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope as an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:99 and the VL sequence of SEQ ID NO:253. In a specific embodiment, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:256 and the VL sequence of SEQ ID NO:253 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope. In a specific embodiment, an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope as an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:256 and the VL sequence of SEQ ID NO:255. In a specific embodiment, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:257 and the VL sequence of SEQ ID NO:195 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope. In a specific embodiment, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:198 and the VL sequence of SEQ ID NO:195 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope.

[0302] In certain embodiments, provided is an Ab-CIDE anti-B7-H4 antibody according to any of the above embodiments that binds to B7-H4 and has at least one of the following characteristics: (a) binds to an epitope within all or part of the B7-H4Ig-V-containing domain (amino acids 29-157 of SEQ ID NO:233); or an epitope within all or part of the B7-H4 Ig-V and Ig-C domains (amino acids 29-250 of SEQ ID NO:233); or an epitope within all or part of SEQ ID NO:234 ​​(mature human B7-H4); or an epitope within all or part of SEQ ID NO:233 (human B7-H4 precursor). In some embodiments, the anti-B7-H4 antibody has one or more of the following characteristics, in any combination: (a) binds to an epitope within all or part of the B7-H4Ig-V-containing domain (amino acids 29-157 of SEQ ID NO:233); or binds to an epitope within all or part of the B7-H4Ig-C-containing domain (amino acids 158-250 of SEQ ID NO:233); Binds epitopes within all or part of the g-V and Ig-C domains (amino acids 29-250 of SEQ ID NO:233); or binds epitopes within all or part of SEQ ID NO:234 ​​(mature human B7-H4); or binds epitopes within all or part of SEQ ID NO:233 (human B7-H4 precursor).

[0303] In a further aspect, the Ab-CIDE anti-B7-H4 antibody according to any of the above embodiments is a monoclonal antibody, including chimeric, humanized, or human antibodies. In one embodiment, the Ab-CIDE anti-B7-H4 antibody is an antibody fragment, e.g., Fv, Fab, Fab', scFv, diabodies, or F(ab') 2 Fragments. In another embodiment, the Ab-CIDE anti-B7-H4 antibody is a substantially full-length antibody, eg, an IgG1 antibody as defined herein, or other antibody class or isotype.

[0304] Antibody 22C10 and other embodiments In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:189, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:190, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:191, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:192, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:193, and (f) the sequence comprising at least 1, 2, 3, 4, 5, or 6 HVRs selected from HVR-L3 comprising the amino acid sequence numbered 194;

[0305] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:218; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:219; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:220; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:221; comprising at least 1, 2, 3, 4, 5, or 6 HVRs selected from HVR-L3 comprising 23 amino acid sequences.

[0306] In one aspect, the Ab-CIDE anti-B7-H4 antibody comprises at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 189, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 190, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 191. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:191. In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:191 and HVR-L3 comprising the amino acid sequence of SEQ ID NO:194. In a further embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:191, HVR-L3 comprising the amino acid sequence of SEQ ID NO:194, and HVR-H2 comprising the amino acid sequence of SEQ ID NO:190. In a further embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 189, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 190, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 191.

[0307] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:218, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:219, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:220. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:220. In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:220 and HVR-L3 comprising the amino acid sequence of SEQ ID NO:223. In a further embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:220, HVR-L3 comprising the amino acid sequence of SEQ ID NO:223, and HVR-H2 comprising the amino acid sequence of SEQ ID NO:219. In a further embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:218, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:219, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:220.

[0308] In another aspect, Ab-CIDE anti-B7-H4 antibodies are provided that comprise at least one, at least two, or all three VL HVR sequences selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 192, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 193, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 194. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 192, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 193, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 194.

[0309] In another aspect, Ab-CIDE anti-B7-H4 antibodies are provided that comprise at least one, at least two, or all three VL HVR sequences selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:221, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:222, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:223. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:221, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:222, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:223.

[0310] In another embodiment, the antibody B7-H4 antibody of Ab-CIDE is a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:189, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:190, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO:191, and (b) (i) the amino acid sequence of SEQ ID NO:192 (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 193; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 194.

[0311] In another embodiment, the antibody B7-H4 antibody of Ab-CIDE is a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:218, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:219, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO:220, and (b) (i) the amino acid sequence of SEQ ID NO:221 (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO:222; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:223.

[0312] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:189; (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:190; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:191; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:192; including HVR-L3 comprising an amino acid sequence selected from 94;

[0313] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:218, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:219, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:220, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:221, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:222, and (f) SEQ ID NO: HVR-L3 comprising an amino acid sequence selected from 223.

[0314] In any of the above embodiments, the Ab-CIDE anti-B7-H4 antibody is a human antibody.

[0315] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:188. In certain embodiments, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 188 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-B7-H4 antibodies comprising that sequence retain the ability to bind B7-H4. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:188. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:188. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, in the FRs). Optionally, the Ab-CIDE anti-B7-H4 antibody comprises the VH sequence of SEQ ID NO: 188, including post-translational modifications of that sequence. In certain embodiments, the VH comprises two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:189, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:190, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:191.

[0316] In another aspect, an Ab-CIDE anti-B7-H4 antibody is provided, wherein the antibody comprises a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:187. In certain embodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 187 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-B7-H4 antibodies comprising that sequence retain the ability to bind B7-H4. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:187. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:187. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, in the FRs). Optionally, the Ab-CIDE anti-B7-H4 antibody comprises the VL sequence of SEQ ID NO: 187, including post-translational modifications of that sequence. In certain embodiments, the VL comprises one, two, or three HVRs selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:192, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:193, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:194.

[0317] In another aspect, an anti-B7-H4 antibody is provided, the antibody comprising a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NOs: 215, 217, 104, 105, or 106. In certain embodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 215, 217, 104, 105, or 106 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, whereas an anti-B7-H4 antibody comprising that sequence -Retains the ability to bind to H4. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:215, 217, 104, 105, or 106. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:215, 217, 104, 105, or 106. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, in the FRs). Optionally, the Ab-CIDE anti-B7-H4 antibody comprises the VL sequence of SEQ ID NO: 215, 217, 104, 105, or 106, including post-translational modifications of that sequence. In certain embodiments, the VL comprises one, two, or three HVRs selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:221, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:222, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:223.

[0318] In another aspect, an Ab-CIDE anti-B7-H4 antibody, wherein the antibody comprises a VH as in any of the embodiments provided above and a VL as in any of the embodiments provided above. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 111 and SEQ ID NO: 104, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 111 and SEQ ID NO: 215, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 112 and SEQ ID NO: 215, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 113 and SEQ ID NO: 215, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 114 and SEQ ID NO: 215, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 111 and SEQ ID NO: 105, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 111 and SEQ ID NO: 106, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 110 and SEQ ID NO: 215, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 109 and SEQ ID NO: 215, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 108 and SEQ ID NO: 215, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 107 and SEQ ID NO: 215, respectively, including post-translational modifications of those sequences. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO:216 and SEQ ID NO:215, respectively, including post-translational modifications of those sequences. In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO:216 and SEQ ID NO:217, respectively, including post-translational modifications of those sequences.

[0319] In a further aspect, provided herein is an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope as the anti-B7-H4 antibody. For example, in certain embodiments, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:188 and the VL sequence of SEQ ID NO:187 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope are provided. In certain embodiments, an Ab-CIDE anti-B7-H4 antibody is provided that binds to the same epitope as an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:111 and the VL sequence of SEQ ID NO:104. In certain embodiments, an Ab-CIDE anti-B7-H4 antibody is provided that binds to the same epitope as an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:111 and the VL sequence of SEQ ID NO:215. In certain embodiments, an Ab-CIDE anti-B7-H4 antibody is provided that binds to the same epitope as an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:112 and the VL sequence of SEQ ID NO:215. In certain embodiments, an Ab-CIDE anti-B7-H4 antibody is provided that binds to the same epitope as an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:113 and the VL sequence of SEQ ID NO:215. In certain embodiments, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:114 and the VL sequence of SEQ ID NO:215 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope are provided. In certain embodiments, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:111 and the VL sequence of SEQ ID NO:105 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope are provided. In certain embodiments, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:111 and the VL sequence of SEQ ID NO:106 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope are provided. In certain embodiments, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:110 and the VL sequence of SEQ ID NO:215 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope are provided. In certain embodiments, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:109 and the VL sequence of SEQ ID NO:215 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope are provided. In a specific embodiment, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:108 and the VL sequence of SEQ ID NO:215 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope. In a specific embodiment, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:107 and the VL sequence of SEQ ID NO:215 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope. In a specific embodiment, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:216 and the VL sequence of SEQ ID NO:215 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope. In a specific embodiment, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:216 and the VL sequence of SEQ ID NO:217 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope.

[0320] In certain embodiments, provided is an Ab-CIDE anti-B7-H4 antibody according to any of the above embodiments that binds to B7-H4 and has at least one of the following characteristics: (a) binds to an epitope within all or part of the B7-H4Ig-V-containing domain (amino acids 29-157 of SEQ ID NO:233); or an epitope within all or part of the B7-H4 Ig-V and Ig-C domains (amino acids 29-250 of SEQ ID NO:233); or an epitope within all or part of SEQ ID NO:234 ​​(mature human B7-H4); or an epitope within all or part of SEQ ID NO:233 (human B7-H4 precursor). In some embodiments, the anti-B7-H4 antibody has one or more of the following characteristics, in any combination: (a) binds to an epitope within all or part of the B7-H4Ig-V-containing domain (amino acids 29-157 of SEQ ID NO:233); or binds to an epitope within all or part of the B7-H4Ig-C-containing domain (amino acids 158-250 of SEQ ID NO:233); Binds epitopes within all or part of the g-V and Ig-C domains (amino acids 29-250 of SEQ ID NO:233); or binds epitopes within all or part of SEQ ID NO:234 ​​(mature human B7-H4); or binds epitopes within all or part of SEQ ID NO:233 (human B7-H4 precursor).

[0321] In a further aspect, the Ab-CIDE anti-B7-H4 antibody according to any of the above embodiments is a monoclonal antibody, including a human antibody. In one embodiment, the Ab-CIDE anti-B7-H4 antibody is an antibody fragment, e.g., Fv, Fab, Fab', scFv, diabodies, or F(ab') 2 Fragments. In another embodiment, the Ab-CIDE anti-B7-H4 antibody is a substantially full-length antibody, eg, an IgG2a antibody as defined herein, or other antibody class or isotype.

[0322] Antibody 32D6 and other embodiments In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:173, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:174, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:175, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:176, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:177, and (f) the sequence comprising at least 1, 2, 3, 4, 5, or 6 HVRs selected from HVR-L3 comprising the amino acid sequence numbered 178;

[0323] In one aspect, the Ab-CIDE anti-B7-H4 antibody comprises at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 173, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 174, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 175. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:175. In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:175 and HVR-L3 comprising the amino acid sequence of SEQ ID NO:178. In further embodiments, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:175, HVR-L3 comprising the amino acid sequence of SEQ ID NO:178, and HVR-H2 comprising the amino acid sequence of SEQ ID NO:174. In a further embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:173, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:174, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:175.

[0324] In another aspect, Ab-CIDE anti-B7-H4 antibodies are provided that comprise at least one, at least two, or all three VL HVR sequences selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 176, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 177, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 178. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 176, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 177, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 178.

[0325] In another embodiment, the antibody B7-H4 antibody of Ab-CIDE is a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:173, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:174, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO:175, and (b) (i) the amino acid sequence of SEQ ID NO:176 (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 177; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 178.

[0326] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 173, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 174, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 175, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 176, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 177, and (f) SEQ ID NO: including HVR-L3 comprising an amino acid sequence selected from 178;

[0327] In any of the above embodiments, the Ab-CIDE anti-B7-H4 antibody is a human antibody.

[0328] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:172. In certain embodiments, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 172 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-B7-H4 antibodies comprising that sequence retain the ability to bind B7-H4. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:172. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:172. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, in the FRs). Optionally, the Ab-CIDE anti-B7-H4 antibody comprises the VH sequence of SEQ ID NO: 172, including post-translational modifications of that sequence. In certain embodiments, the VH comprises two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:173, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:174, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:175.

[0329] In another aspect, an Ab-CIDE anti-B7-H4 antibody is provided, wherein the antibody comprises a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:171. In certain embodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 171 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-B7-H4 antibodies comprising that sequence retain the ability to bind B7-H4. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:171. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:171. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, in the FRs). Optionally, the Ab-CIDE anti-B7-H4 antibody comprises the VL sequence of SEQ ID NO: 171, including post-translational modifications of that sequence. In certain embodiments, the VL comprises one, two, or three HVRs selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:176, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:177, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:178.

[0330] In another aspect, an Ab-CIDE anti-B7-H4 antibody, wherein the antibody comprises a VH as in any of the embodiments provided above and a VL as in any of the embodiments provided above. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 172 and SEQ ID NO: 171, respectively, including post-translational modifications of those sequences. In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 172 and SEQ ID NO: 171, respectively, including post-translational modifications of those sequences.

[0331] In a further aspect, provided herein is an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope as the anti-B7-H4 antibody. For example, in certain embodiments, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:172 and the VL sequence of SEQ ID NO:171 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope.

[0332] In certain embodiments, provided is an Ab-CIDE anti-B7-H4 antibody according to any of the above embodiments that binds to B7-H4 and has at least one of the following characteristics: (a) binds to an epitope within all or part of the B7-H4Ig-V-containing domain (amino acids 29-157 of SEQ ID NO:233); or an epitope within all or part of the B7-H4 Ig-V and Ig-C domains (amino acids 29-250 of SEQ ID NO:233); or an epitope within all or part of SEQ ID NO:234 ​​(mature human B7-H4); or an epitope within all or part of SEQ ID NO:233 (human B7-H4 precursor). In some embodiments, the anti-B7-H4 antibody has one or more of the following characteristics, in any combination: (a) binds to an epitope within all or part of the B7-H4Ig-V-containing domain (amino acids 29-157 of SEQ ID NO:233); or binds to an epitope within all or part of the B7-H4Ig-C-containing domain (amino acids 158-250 of SEQ ID NO:233); binds epitopes within all or part of the g-V and Ig-C domains (amino acids 29-250 of SEQ ID NO:233); or binds epitopes within all or part of SEQ ID NO:234 ​​(mature human B7-H4); binds epitopes within all or part of SEQ ID NO:233 (human B7-H4 precursor).

[0333] In a further aspect, the Ab-CIDE anti-B7-H4 antibody according to any of the above embodiments is a monoclonal antibody, including a human antibody. In one embodiment, the Ab-CIDE anti-B7-H4 antibody is an antibody fragment, e.g., Fv, Fab, Fab', scFv, diabodies, or F(ab')2 Fragments. In another embodiment, the Ab-CIDE anti-B7-H4 antibody is a substantially full-length antibody, eg, an IgG2a antibody as defined herein, or other antibody class or isotype.

[0334] Antibody 9B9 and other embodiments In some embodiments, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:181, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:182, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:183, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:184, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:185, and (f) the sequence comprising at least 1, 2, 3, 4, 5, or 6 HVRs selected from HVR-L3 comprising the amino acid sequence numbered 186;

[0335] In one aspect, the Ab-CIDE anti-B7-H4 antibody comprises at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 181, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 182, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 183. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:183. In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:183 and HVR-L3 comprising the amino acid sequence of SEQ ID NO:186. In a further embodiment, the Ab-CIDE anti-B7-H4 antibody comprises HVR-H3 comprising the amino acid sequence of SEQ ID NO:183, HVR-L3 comprising the amino acid sequence of SEQ ID NO:186, and HVR-H2 comprising the amino acid sequence of SEQ ID NO:182. In a further embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:181, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:182, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:183.

[0336] In another aspect, Ab-CIDE anti-B7-H4 antibodies are provided that comprise at least one, at least two, or all three VL HVR sequences selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 184, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 185, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 186. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 184, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 185, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 186.

[0337] In another embodiment, the antibody B7-H4 antibody of Ab-CIDE is a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:181, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:182, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO:183, and (b) (i) the amino acid sequence of SEQ ID NO:184 (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 185; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 186.

[0338] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 181, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 182, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 183, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 184, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 185, and (f) SEQ ID NO: including HVR-L3 comprising an amino acid sequence selected from 186;

[0339] In any of the above embodiments, the Ab-CIDE anti-B7-H4 antibody is a human antibody.

[0340] In another embodiment, the Ab-CIDE anti-B7-H4 antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:180. In certain embodiments, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 180 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-B7-H4 antibodies comprising that sequence retain the ability to bind B7-H4. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:180. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:180. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, in the FRs). Optionally, the Ab-CIDE anti-B7-H4 antibody comprises the VH sequence of SEQ ID NO: 180, including post-translational modifications of that sequence. In certain embodiments, the VH comprises two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:181, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:182, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:183.

[0341] In another aspect, an anti-B7-H4 antibody is provided, the antibody comprising a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:179. In certain embodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO: 179 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-B7-H4 antibodies comprising that sequence retain the ability to bind B7-H4. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:179. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:179. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, in the FRs). Optionally, the Ab-CIDE anti-B7-H4 antibody comprises the VL sequence of SEQ ID NO: 171, including post-translational modifications of that sequence. In certain embodiments, the VL comprises 1, 2, or 3 HVRs selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 184, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 185, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 186.

[0342] In another aspect, an Ab-CIDE anti-B7-H4 antibody, wherein the antibody comprises a VH as in any of the embodiments provided above and a VL as in any of the embodiments provided above. In one embodiment, the Ab-CIDE anti-B7-H4 antibody comprises the VH and VL sequences of SEQ ID NO: 180 and SEQ ID NO: 179, respectively, including post-translational modifications of those sequences.

[0343] In a further aspect, provided herein is an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope as the anti-B7-H4 antibody. For example, in certain embodiments, an anti-B7-H4 antibody comprising the VH sequence of SEQ ID NO:180 and the VL sequence of SEQ ID NO:179 and an Ab-CIDE anti-B7-H4 antibody that binds to the same epitope are provided.

[0344] In certain embodiments, provided is an Ab-CIDE anti-B7-H4 antibody to B7-H4 according to any of the above embodiments that has at least one of the following characteristics: (a) binds to an epitope within all or part of the B7-H4Ig-V containing domain (amino acids 29-157 of SEQ ID NO:233); or within all or part of the B7-H4Ig-C containing domain (amino acids 158-250 of SEQ ID NO:233). or within all or part of the B7-H4 Ig-V and Ig-C domains (amino acids 29-250 of SEQ ID NO:233); or within all or part of SEQ ID NO:234 ​​(mature human B7-H4); or within all or part of SEQ ID NO:233 (human B7-H4 precursor); , in any combination, having one or more of the following characteristics: (a) binds to an epitope within all or part of the B7-H4Ig-V-containing domain (amino acids 29-157 of SEQ ID NO:233); or binds to an epitope within all or part of the B7-H4Ig-C-containing domain (amino acids 158-250 of SEQ ID NO:233); or epitopes within all or part of the B7-H4Ig-V and Ig-C domains. Binds a tope (amino acids 29-250 of SEQ ID NO:233); or binds an epitope within all or part of SEQ ID NO:234 ​​(mature human B7-H4); or binds an epitope within all or part of SEQ ID NO:233 (human B7-H4 precursor).

[0345] In a further aspect, the Ab-CIDE anti-B7-H4 antibody according to any of the above embodiments is a monoclonal antibody, including a human antibody. In one embodiment, the Ab-CIDE anti-B7-H4 antibody is an antibody fragment, e.g., Fv, Fab, Fab', scFv, diabodies, or F(ab') 2 Fragments. In another embodiment, the Ab-CIDE anti-B7-H4 antibody is a substantially full-length antibody, eg, an IgG2a antibody as defined herein, or other antibody class or isotype.

[0346] Anti-MUC16 antibody In certain embodiments, Ab-CIDE comprises an anti-MUC16 antibody.

[0347] (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:36; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:37; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:32; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:33; A PAC comprising an anti-MUC16 antibody comprising at least 1, 2, 3, 4, 5, or 6 HVRs is described.

[0348] In one aspect, described herein is an Ab-CIDE comprising an antibody comprising at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:35, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:36, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:37. In further embodiments, the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:35, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:36, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:37.

[0349] In another aspect, described herein is an Ab-CIDE comprising an antibody comprising at least one, at least two, or all three VL HVR sequences selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:32, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:33, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:34. In one embodiment, the antibody comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:32, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:33, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:34.

[0350] In another embodiment, the Ab-CIDE is a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:35, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:36, and (iii) HVR-H3 comprising an amino acid sequence selected from SEQ ID NO:37, and (b) (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO:32, (ii) a) HVR-L2 comprising the amino acid sequence of SEQ ID NO:33; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:34.

[0351] In another aspect, provided herein is an antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:35, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:36, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:37, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:32, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:33, and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:34. Ab-CIDE is described, including

[0352] In any of the above embodiments, the Ab-CIDE anti-MUC16 antibody is humanized. In one embodiment, the anti-MUC16 antibody comprises an HVR as in any of the above embodiments and further comprises a human acceptor framework, eg a human immunoglobulin framework or a human consensus framework.

[0353] In another embodiment, the Ab-CIDE anti-MUC16 antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:39. In certain embodiments, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:39 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-MUC16 antibodies comprising that sequence retain the ability to bind MUC16. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:39. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:39. In certain embodiments, substitutions, insertions, or deletions occur within regions outside the HVR (ie, within FRs). Optionally, the anti-MUC16 antibody comprises the VH sequence of SEQ ID NO:39, including post-translational modifications of that sequence. In certain embodiments, the VH comprises two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:35, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:36, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:37.

[0354] In another aspect, Ab-CIDE anti-MUC16 antibodies are provided, said antibodies comprising a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:38. In certain embodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:38 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-MUC16 antibodies comprising the sequence retain the ability to bind MUC16. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:38. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:38. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, within the FRs). Optionally, the anti-MUC16 antibody comprises the VL sequence of SEQ ID NO:38, including post-translational modifications of that sequence. In certain embodiments, the VL comprises one, two, or three HVRs selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:32, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:33, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:34.

[0355] In another aspect there is provided an Ab-CIDE comprising an anti-MUC16 antibody, the antibody comprising a VH as in any of the above embodiments and a VL as in any of the above embodiments.

[0356] In one embodiment, an Ab-CIDE is provided, the antibody comprising the VH and VL sequences of SEQ ID NO:39 and SEQ ID NO:38, respectively, including post-translational modifications of those sequences.

[0357] In a further aspect, provided herein is an Ab-CIDE comprising an antibody that binds to the same epitope as an anti-MUC16 antibody provided herein. For example, in certain embodiments, a PAC is provided that includes an antibody that binds to the same epitope as an anti-MUC16 antibody that includes the VH sequence of SEQ ID NO:39 and the VL sequence of SEQ ID NO:38, respectively.

[0358] In a further aspect, the Ab-CIDE anti-MUC16 antibody according to any of the above embodiments is a monoclonal antibody, including a human antibody. In one embodiment, the Ab-CIDE anti-MUC16 antibody is an antibody fragment, e.g., Fv, Fab, Fab', scFv, diabodies, or F(ab') 2 Fragments. In another embodiment, the antibody is a substantially full-length antibody, eg, an IgG1 antibody, IgG2a antibody, or other antibody class or isotype as defined herein.

[0359] Anti-STEAP-1 antibody In certain embodiments, Ab-CIDE comprises an anti-STEAP-1 antibody.

[0360] In some embodiments, selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:40, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:41, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:42, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:43, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:44, and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:45 A PAC comprising an anti-STEAP-1 antibody is described that comprises at least 1, 2, 3, 4, 5, or 6 HVRs.

[0361] In one aspect, described herein is an Ab-CIDE comprising an antibody comprising at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:40, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:41, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:42. In further embodiments, the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:40, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:41, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:42.

[0362] In another aspect, described herein is an Ab-CIDE comprising an antibody comprising at least one, at least two, or all three VL HVR sequences selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:43, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:44, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:45. In one embodiment, the antibody comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:43, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:44, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:45.

[0363] In another embodiment, the Ab-CIDE is a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:40, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:41, and (iii) HVR-H3 comprising an amino acid sequence selected from SEQ ID NO:42, and (b) (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO:43, (ii) a) HVR-L2 comprising the amino acid sequence of SEQ ID NO:44; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:45.

[0364] In another aspect, provided herein is an antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:40, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:41, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:42, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:43, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:44, and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:45. Ab-CIDE is described, including

[0365] In any of the above embodiments, the Ab-CIDE anti-STEAP-1 antibody is humanized. In one embodiment, the anti-STEAP-1 antibody comprises an HVR as in any of the above embodiments and further comprises a human acceptor framework, eg a human immunoglobulin framework or a human consensus framework.

[0366] In another embodiment, the Ab-CIDE anti-STEAP-1 antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:46. In certain embodiments, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:46 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-STEAP-1 antibodies containing that sequence retain the ability to bind STEAP-1. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:46. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:46. In certain embodiments, substitutions, insertions, or deletions occur within regions outside the HVR (ie, within FRs). Optionally, the anti-STEAP-1 antibody comprises the VH sequence of SEQ ID NO:46, including post-translational modifications of that sequence. In certain embodiments, the VH comprises two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:40, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:41, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:42.

[0367] In another aspect, Ab-CIDE anti-STEAP-1 antibodies are provided, said antibodies comprising a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:47. In certain embodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:47 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-STEAP-1 antibodies comprising that sequence retain the ability to bind STEAP-1. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted within SEQ ID NO:47, and in certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted within SEQ ID NO:47. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, within the FRs). Optionally, the anti-STEAP-1 antibody comprises the VL sequence of SEQ ID NO:47, including post-translational modifications of that sequence. In certain embodiments, the VL comprises one, two, or three HVRs selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:43, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:44, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:45.

[0368] In another aspect there is provided an Ab-CIDE comprising an anti-STEAP-1 antibody, the antibody comprising a VH as in any of the above embodiments and a VL as in any of the above embodiments.

[0369] In one embodiment, an Ab-CIDE is provided, the antibody comprising the VH and VL sequences of SEQ ID NO:46 and SEQ ID NO:47, respectively, including post-translational modifications of those sequences.

[0370] In a further aspect, provided herein is an Ab-CIDE comprising an antibody that binds to the same epitope as an anti-STEAP-1 antibody provided herein. For example, in certain embodiments, an Ab-CIDE is provided comprising an antibody that binds to the same epitope as an anti-STEAP-1 antibody comprising the VH sequence of SEQ ID NO:46 and the VL sequence of SEQ ID NO:47, respectively.

[0371] In a further aspect, the Ab-CIDE anti-STEAP-1 antibody according to any of the above embodiments is a monoclonal antibody, including a human antibody. In one embodiment, the Ab-CIDE anti-STEAP-1 antibody is an antibody fragment, e.g., Fv, Fab, Fab', scFv, diabodies, or F(ab') 2 Fragments. In another embodiment, the antibody is a substantially full-length antibody, eg, an IgG1 antibody, IgG2a antibody, or other antibody class or isotype as defined herein.

[0372] Anti-NaPi2b antibody In certain embodiments, Ab-CIDE comprises an anti-NaPi2b antibody.

[0373] (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:49; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:50; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:51; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:52; Ab-CIDEs comprising anti-NaPi2b antibodies comprising at least 1, 2, 3, 4, 5, or 6 HVRs are described.

[0374] In one aspect, described herein is an Ab-CIDE comprising an antibody comprising at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:48, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:49, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:50. In further embodiments, the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:48, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:49, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:50.

[0375] In another aspect, described herein is an Ab-CIDE comprising an antibody comprising at least one, at least two, or all three VL HVR sequences selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:51, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:52, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:53. In one embodiment, the antibody comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:51, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:52, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:53.

[0376] In another embodiment, the Ab-CIDE is a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:48, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:49, and (iii) HVR-H3 comprising an amino acid sequence selected from SEQ ID NO:50, and (b) (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO:51, (ii) a) HVR-L2 comprising the amino acid sequence of SEQ ID NO:52; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:53.

[0377] In another aspect, provided herein is an antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:48, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:49, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:50, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:51, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:52, and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:53. Ab-CIDE is described, including

[0378] In any of the above embodiments, the Ab-CIDE anti-NaPi2b antibody is humanized. In one embodiment, the anti-NaPi2b antibody comprises an HVR as in any of the above embodiments and further comprises a human acceptor framework, eg a human immunoglobulin framework or a human consensus framework.

[0379] In another embodiment, the Ab-CIDE anti-NaPi2b antibody comprises a heavy chain variable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:54. In certain embodiments, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:54 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-NaPi2b antibodies comprising that sequence retain the ability to bind NaPi2b. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:54. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:54. In certain embodiments, substitutions, insertions, or deletions occur within regions outside the HVR (ie, within FRs). Optionally, the anti-NaPi2b antibody comprises the VH sequence of SEQ ID NO:54, including post-translational modifications of that sequence. In certain embodiments, the VH comprises two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:48, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:49, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:50.

[0380] In another aspect, an Ab-CIDE anti-NaPi2b antibody is provided, the antibody comprising a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:55. In certain embodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:55 contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but anti-NaPi2b antibodies comprising that sequence retain the ability to bind anti-NaPi2b. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:55. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:55. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, within the FRs). Optionally, the anti-NaPi2b antibody comprises the VL sequence of SEQ ID NO:55, including post-translational modifications of that sequence. In certain embodiments, the VL comprises one, two, or three HVRs selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:51, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:52, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:53.

[0381] In another aspect there is provided an Ab-CIDE comprising an anti-NaPi2b antibody, the antibody comprising a VH as in any of the above embodiments and a VL as in any of the above embodiments.

[0382] In one embodiment, an Ab-CIDE is provided, the antibody comprising the VH and VL sequences of SEQ ID NO:54 and SEQ ID NO:55, respectively, including post-translational modifications of those sequences.

[0383] In a further aspect, provided herein is an Ab-CIDE comprising an antibody that binds to the same epitope as an anti-NaPi2b antibody provided herein. For example, in certain embodiments, Ab-CIDE is provided comprising an antibody that binds to the same epitope as an anti-NaPi2b antibody comprising the VH sequence of SEQ ID NO:54 and the VL sequence of SEQ ID NO:55, respectively.

[0384] In a further aspect, the Ab-CIDE anti-NaPi2b antibody according to any of the above embodiments is a monoclonal antibody, including a human antibody. In one embodiment, the Ab-CIDE anti-NaPi2b antibody is an antibody fragment such as Fv, Fab, Fab', scFv, diabodies, or F(ab') 2 Fragments. In another embodiment, the antibody is a substantially full-length antibody, eg, an IgG1 antibody, IgG2a antibody, or other antibody class or isotype as defined herein.

[0385] Anti-CD79b antibody In certain embodiments, Ab-CIDE comprises an anti-CD79b antibody.

[0386] (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:59; (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:60; (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:61; (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:62; Ab-CIDEs comprising anti-CD79b antibodies comprising at least 1, 2, 3, 4, 5, or 6 HVRs are described.

[0387] In one aspect, described herein is an Ab-CIDE comprising an antibody comprising at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:58, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:59, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:60. In further embodiments, the antibody comprises (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:58, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:59, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:60.

[0388] In another aspect, described herein is an Ab-CIDE comprising an antibody comprising at least one, at least two, or all three VL HVR sequences selected from (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:61, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:62, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:63. In one embodiment, the antibody comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:61, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:62, and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:63.

[0389] In another embodiment, the Ab-CIDE is a VH domain comprising at least one, at least two, or all three VH HVR sequences selected from (a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO:58, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO:59, and (iii) HVR-H3 comprising an amino acid sequence selected from SEQ ID NO:60, and (b) (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO:61, (ii) a) HVR-L2 comprising the amino acid sequence of SEQ ID NO:62; and (c) HVR-L3 comprising the amino acid sequence of SEQ ID NO:63.

[0390] In another aspect, provided herein is an antibody comprising (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:58, (b) HVR-H2 comprising the amino acid sequence of SEQ ID NO:59, (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:60, (d) HVR-L1 comprising the amino acid sequence of SEQ ID NO:61, (e) HVR-L2 comprising the amino acid sequence of SEQ ID NO:62, and (f) HVR-L3 comprising the amino acid sequence of SEQ ID NO:63. Ab-CIDE is described, including

[0391] In any of the above embodiments, the Ab-CIDE anti-CD79b antibody is humanized. In one embodiment, the anti-CD79b antibody comprises an HVR as in any of the above embodiments and further comprises a human acceptor framework, eg a human immunoglobulin framework or a human consensus framework.

[0392] In another embodiment, the Ab-CIDE anti-CD79b antibody is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% relative to the amino acid sequence of SEQ ID NO:56 , 99%, or 100% sequence identity. In certain embodiments, at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:56 A VH sequence having a sequence contains substitutions (eg, conservative substitutions), insertions, or deletions compared to a reference sequence, yet an anti-CD79b antibody containing that sequence retains the ability to bind CD79b. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:56. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:56. In certain embodiments, substitutions, insertions, or deletions occur within regions outside the HVR (ie, within FRs). Optionally, the anti-CD79b antibody comprises the VH sequence of SEQ ID NO:8, including post-translational modifications of that sequence. In certain embodiments, the VH comprises two or three HVRs selected from: (a) HVR-H1 comprising the amino acid sequence of SEQ ID NO:58, (b) HVR comprising the amino acid sequence of SEQ ID NO:59 -H2, and (c) HVR-H3 comprising the amino acid sequence of SEQ ID NO:60.

[0393] In another aspect, an Ab-CIDE anti-CD79b antibody is provided, wherein the antibody is at least 90%, 91%, 92%, 93%, 94%, 95%, 96% A light chain variable domain (VL) with %, 97%, 98%, 99% or 100% sequence identity. In certain embodiments, at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence of SEQ ID NO:57 A VL string with a sequence contains substitutions (eg, conservative substitutions), insertions, or deletions compared to the reference sequence, but anti-Ly6E antibodies containing that sequence retain the ability to bind CD79b. In certain embodiments, a total of 1-10 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:57. In certain embodiments, a total of 1-5 amino acids have been substituted, inserted, and / or deleted in SEQ ID NO:57. In certain embodiments, substitutions, insertions, or deletions occur in regions outside the HVR (ie, within the FRs). Optionally, the anti-CD79b antibody comprises the VL sequence of SEQ ID NO:57, including post-translational modifications of that sequence. In certain embodiments, the VL comprises (a) HVR-L1 comprising the amino acid sequence of SEQ ID NO:61, (b) HVR-L2 comprising the amino acid sequence of SEQ ID NO:62, and (c) the amino acid sequence of SEQ ID NO:63. Contains 1, 2, or 3 HVRs selected from HVR-L3, including.

[0394] In another aspect, described herein and provided is an Ab-CIDE comprising an anti-CD79b antibody, wherein the antibody comprises a VH as in any of the above embodiments and an anti-CD79b antibody in any of the above embodiments. Including VL as in

[0395] In one embodiment, an Ab-CIDE is provided, the antibody comprising the VH and VL sequences of SEQ ID NO:56 and SEQ ID NO:57, respectively, including post-translational modifications of those sequences.

[0396] In a further aspect, provided herein is an Ab-CIDE comprising an antibody that binds to the same epitope as an anti-CD79b antibody provided herein. For example, in certain embodiments, Ab-CIDE is provided comprising an antibody that binds to the same epitope as an anti-CD79b antibody comprising the VH sequence of SEQ ID NO:56 and the VL sequence of SEQ ID NO:57, respectively.

[0397] In a further aspect, the Ab-CIDE anti-CD79b antibody according to any of the above embodiments is a monoclonal antibody, including a human antibody. In one embodiment, the Ab-CIDE anti-CD79b antibody is an antibody fragment such as Fv, Fab, Fab', scFv, diabodies, or F(ab') 2 Fragments. In another embodiment, the antibody is a substantially full-length antibody, eg, an IgG1 antibody, IgG2a antibody, or other antibody class or isotype as defined herein.

[0398] Anti-CD22 antibody In certain embodiments, the Ab-CIDE comprises three light chain hypervariable regions (HVR-L1, HVR-L2 and HVR-L3) and three heavy chain hypervariable regions (HVR-H1, HVR-H2 and HVR- H3), including anti-CD22 antibodies. In one embodiment, the Ab-CIDE anti-CD22 antibody comprises three light chain hypervariable regions and three heavy chain hypervariable regions (SEQ ID NOS:66-71), the sequences of which are shown below. In one embodiment, the Ab-CIDE anti-CD22 antibody comprises the variable light chain sequence of SEQ ID NO:72 and the variable heavy chain sequence of SEQ ID NO:73. In one embodiment, the Ab-CIDEs anti-CD22 antibody of the invention comprises the light chain sequence of SEQ ID NO:74 and the heavy chain sequence of SEQ ID NO:75:

[0399] Anti-CD33 antibody In certain embodiments, the Ab-CIDE can comprise an anti-CD33 antibody comprising 3 light chain hypervariable regions and 3 heavy chain hypervariable regions, the sequences of which (SEQ ID NOs:76-81) are shown below. . In one embodiment, the Ab-CIDE anti-CD33 antibody comprises the variable light chain sequence of SEQ ID NO:82 and the variable heavy chain sequence of SEQ ID NO:83.

[0400] In one embodiment, the Ab-CIDE anti-CD33 antibody comprises the light chain sequence of SEQ ID NO:84 and the heavy chain sequence of SEQ ID NO:85. In one embodiment, the Ab-CIDE anti-CD33 antibody comprises three light chain hypervariable regions and three heavy chain hypervariable regions, the sequences of which (SEQ ID NOS:84-89) are shown below. In one embodiment, the Ab-CIDE anti-CD33 antibody comprises the variable light chain sequence of SEQ ID NO:90 and the variable heavy chain sequence of SEQ ID NO:91. In one embodiment, the Ab-CIDE anti-CD33 antibody comprises the variable light chain sequence of SEQ ID NO:92 and the variable heavy chain sequence of SEQ ID NO:93. In one embodiment, an anti-CD33 antibody of the invention comprises the variable light chain sequence of SEQ ID NO:94 and the variable heavy chain sequence of SEQ ID NO:95. In one embodiment, an anti-CD33 antibody of the invention comprises the variable light chain sequence of SEQ ID NO:96 and the variable heavy chain sequence of SEQ ID NO:97.

[0401] 1. Antibody affinity In certain embodiments, the antibodies provided herein have a dissociation constant ( Kd), optionally ≥10 -13 M (e.g. 10 -8 M or less, e.g. 10 -8 M~10 -13 M, for example 10 -9 M~10 -13 M).

[0402] In one embodiment, the Kd is measured by a radiolabeled antigen binding assay (RIA) performed using the Fab version of the antibody of interest and its antigen, as described by the assay below. Solution binding affinities of Fabs for antigen were evaluated in the presence of serial titrations of unlabeled antigen at a minimal concentration of Fabs ( 125 I) Measured by equilibrating with labeled antigen and then capturing bound antigen with an anti-Fab antibody-coated plate (e.g., Chen et al., J. Mol. Biol. 293:865-881 (1999). )). To establish assay conditions, MICROTITER® multiwell plates (Thermo Scientific) were coated overnight with 5 μg / ml of capture anti-Fab antibody (Cappel Labs) in 50 mM sodium carbonate (pH 9.6). followed by blocking with 2% (w / v) bovine serum albumin in PBS for 2-5 hours at room temperature (approximately 23°C). 100 pM or 26 pM [ 125 I]-antigen is mixed with serial dilutions of the Fab of interest (e.g., evaluation of the anti-VEGF antibody Fab-12 in Presta et al., Cancer Res. 57:4593-4599 (1997)). consistent with). The Fab of interest is then incubated overnight, although this incubation may continue for a longer period (eg, about 65 hours) to ensure equilibrium is reached. The mixture is then transferred to a capture plate for incubation (eg, 1 hour) at room temperature. The solution is then removed and the plate washed 8 times with 0.1% polysorbate 20 (TWEEN-20®) in PBS. Once the plates are dry, add 150 μL / well of scintillant (MICROSCINT-20 (商標) , Packard) and plate the topcount (商標) Count for 10 minutes on a gamma counter (Packard). Concentrations of each Fab that yield 20% or less of maximal binding are selected for use in competitive binding assays.

[0403] According to another embodiment, the Kd is measured using a surface plasmon resonance assay using a BIACORE®-2000 or BIACORE®-3000 (BIAcore, Inc., Piscataway, NJ) at 25° C. with antigen CM5 chips immobilized at about 10 response units (RU). Briefly, a carboxymethylated dextran biosensor chip (CM5, BIACORE, Inc.) is activated with N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) according to the supplier's instructions. Antigen is diluted to 5 μg / ml (approximately 0.2 μM) with 10 mM sodium acetate pH 4.8 before injection at a flow rate of 5 μl / min to achieve approximately 10 response units (RU) of coupled protein. After injection of antigen, 1M ethanolamine is injected to block unreacted groups. For kinetic measurements, two-fold serial dilutions of Fabs (0.78 nM to 500 nM) are injected in PBS with 0.05% polysorbate 20 (TWEEN-20™) surfactant (PBST) at 25°C at a flow rate of approximately 25 μL / min. Association rates (kon) and dissociation rates (koff) are calculated using a simple one-to-one Langmuir binding model (BIACORE® Evaluation Software version 3.2) by fitting the association and dissociation sensograms simultaneously. The equilibrium dissociation constant (Kd) is calculated as the koff / kon ratio. See, eg, Chen et al., J. Mol. Biol., 293:865-881 (1999). Fluorescence emission intensity of 20 nM anti-antigen antibody (Fab form) in PBS (pH 7.2) at 25°C in the presence of increasing concentrations of antigen, as measured by a spectrophotometer such as the 8000 series SLM-AMINCO™ spectrophotometer (ThermoSpectronic) with a stop-flow equipped spectrophotometer (Aviv Instruments) or with a stirring cuvette, when the on-rate exceeds 10 M s by the surface plasmon resonance assay described above. (Excitation = 295 nm, Emission = 340 nm, 16 nm bandpass) can be determined by using a fluorescence quenching technique that measures the increase or decrease.

[0404] 2. Linker (L1) A “linker” (L1, linker-1), as described herein, is a bifunctional or multifunctional moiety that can be used to link one or more CIDE moieties (D) to an antibody (Ab) to form Ab-CIDE. In some embodiments, Ab-CIDE can be prepared using L1 with reactive functional groups for covalent conjugation to CIDE and to antibodies. For example, in some embodiments, a cysteine ​​thiol of an antibody (Ab) can form a bond with a reactive functional group of a linker or a linker L1-CIDE group to create Ab-CIDE. Specifically, the chemical structure of the linker can have a large impact on both the efficacy and safety of Ab-CIDE (Ducry & Stump, "Bioconjugate Chem", 2010, 21, 5-13). Choosing the correct linker affects proper drug delivery to the intended cellular compartment of the target cell.

[0405] Linkers can generally be divided into two categories: cleavable (eg, peptides, hydrazones or disulfides, etc.) or non-cleavable (thioethers, etc.). If the linker is a non-cleavable linker, its position on the E3LB portion is such that it does not interfere with VHL binding. Specifically, the non-cleavable linker should not be covalently attached at the hydroxyl position on the proline of the VHL binding domain. Peptide linkers such as valine-citrulline (Val-Cit), which can be hydrolyzed by lysosomal enzymes (such as cathepsin B), have been used to link drugs to antibodies (US Pat. No. 6,214,345). They were particularly useful due in part to their relative stability in the systemic circulation and ability to efficiently release drugs within tumors. However, due to the limited chemical space represented by natural peptides, it is desirable to have a variety of non-peptide linkers that behave like peptides and can be effectively cleaved by lysosomal proteases. Greater non-peptide structural diversity can lead to novel beneficial properties not conferred by peptide linkers. Provided herein are different types of non-peptide linkers for linker L1 that can be cleaved by lysosomal enzymes.

[0406] a peptidomimetic linker Provided herein are different types of non-peptidic peptidomimetic linkers for Ab-CIDE that are cleavable by lysosomal enzymes. For example, the central amide bond of a dipeptide (e.g., Val-Cit) has been replaced with an amide mimetic, and / or the entire amino acid (e.g., the valine amino acid in the Val-Cit dipeptide) has been replaced with a non-amino acid moiety (e.g., a cycloalkyldicarbonyl structure (e.g., ring size = 4 or 5)).

[0407] When L1 is a peptidomimetic linker, the formula: -Str-(PM)-Sp-, In the formula: Str is the stretcher unit covalently attached to Ab; Sp is a spacer unit covalently attached to a bond or CIDE moiety;; and PM is TIFF2023100643000001.tif107170 In the formula, W is -NH-heterocycloalkyl- or heterocycloalkyl; Y is heteroaryl, aryl, -C(O)C 1 ~C 6 Alkylene, C 1 ~C 6 Alkylene-NH 2 , C 1 ~C 6 Alkylene-NH-CH 3 , C 1 ~C 6 Alkylene-N-(CH 3 ) 2 , C 1 ~C 6 alkenyl, or C 1 ~C 6 is alkylenyl; Each R 1 independently, C 1 ~C 10 Alkyl, C 1 ~C 10 alkenyl, (C 1 ~C 10 Alkyl)NHC(NH)NH 2 , or (C 1 ~C 10 Alkyl)NHC(O)NH 2 is; R. 3 and R 2 are independently H, C 1 ~C 10 Alkyl, C 1 ~C 10 alkenyl, arylalkyl, or heteroarylalkyl, or R 3 and R 2 together, C 3 ~C 7 may form a cycloalkyl; and R. 4 and R 5 are each independently C 1 ~C 10 Alkyl, C 1 ~C 10 alkenyl, arylalkyl, heteroarylalkyl, (C 1 ~C 10 Alkyl)OCH 2 - or R 4 and R 5 is C 3 ~C 7 may form a cycloalkyl ring, non-peptide chemical moieties selected from the group consisting of represented by

[0408] Note that L1 may be connected to CIDE through either the E3LB, L2, or PB groups.

[0409] In embodiments, Y is heteroaryl and R 4 and R 5 together form a cyclobutyl ring.

[0410] In embodiments, Y is a moiety selected from the group consisting of: TIFF2023100643000002.tif23170

[0411] In embodiments, Str is a chemical moiety represented by the formula: TIFF2023100643000003.tif34170 R 6 is C 1 ~C 10 Alkylene, C 1 ~C 10 alkenyl, C 3 ~C 8 cycloalkyl, (C 1 ~C 8 alkylene)O-, and C 1 ~C 10 Alkylene-C(O)N(R a )-C 2 ~C 6is selected from the group consisting of alkylene, each alkylene being halo, trifluoromethyl, difluoromethyl, amino, alkylamino, cyano, sulfonyl, sulfonamide, sulfoxide, hydroxy, alkoxy, ester, carboxylic acid, alkylthio, C 3 ~C 8 Cycloalkyl, C 4 ~C 7 optionally substituted with 1 to 5 substituents selected from the group consisting of heterocycloalkyl, heteroarylalkyl, arylarylalkyl, heteroarylalkyl and heteroaryl, each R a is independently H or 1 ~C 6 is alkyl; Sp is -Ar-R b -, Ar is aryl or heteroaryl, and R b is (C 1 ~C 10 alkylene)O-.

[0412] In one exemplary embodiment, Str has the formula: TIFF2023100643000004.tif23170 formula, R 7 is C 1 ~C 10 Alkylene, C 1 ~C 10 alkenyl, (C 1 ~C 10 Alkylene)O-, N(R c )-(C 2 ~C 6 Alkylene)-N(R c ) and N(R c )-(C 2 ~C 6 alkylene) and each R c is independently H or C 1 ~C 6 is alkyl; Sp is -Ar-R b -, Ar is aryl or heteroaryl, and R b is (C 1 ~C 10 Alkylene)O- or Sp-C 1 ~C 6 -alkylene-C(O)NH-.

[0413] In embodiments, L1 is a non-peptide chemical moiety represented by the formula: TIFF2023100643000005.tif28170R 1 is C 1 ~C 6 Alkyl, C 1 ~C 6 alkenyl, (C 1 ~C 6 Alkyl)NHC(NH)NH 2 or (C 1 ~C 6 Alkyl)NHC(O)NH 2 is; R. 3 and R 2 are each independently H or C 1 ~C 10 is alkyl.

[0414] In embodiments, L1 is a non-peptide chemical moiety represented by the formula: TIFF2023100643000006.tif25170R 1 is C 1 ~C 6 alkyl, (C 1 ~C 6 Alkyl)NHC(NH)NH 2 , or (C 1 ~C 6 Alkyl)NHC(O)NH 2 is; R. 4 and R 5 together, C 3 ~C 7 Forms a cycloalkyl ring.

[0415] In embodiments, L1 is a non-peptide chemical moiety represented by the formula: TIFF2023100643000007.tif30170R 1 is C 1 ~C 6 Alkyl, C 1 ~C 6 Alkyl)NHC(NH)NH 2 or (C 1 ~C 6 Alkyl)NHC(O)NH 2 and W is as defined above.

[0416] In some embodiments, the linker can be a peptidomimetic linker such as those described in WO2015 / 095227, WO2015 / 095124 or WO2015 / 095223.

[0417] In certain embodiments, the linker is selected from the group consisting of: TIFF2023100643000008.tif92170

[0418] b. non-peptidomimetic linker In one aspect, linker L1 forms a disulfide bond with the antibody. In one aspect, the linker has the structure: TIFF2023100643000009.tif91170In formula, R 1 and R 2 are independently H, and C 1 ~C 6 independently selected from alkyl, or R 1 and R 2 forms a 3-, 4-, 5-, or 6-membered cycloalkyl or heterocyclic group. A linker can be covalently attached to the antibody and CIDE as follows. TIFF2023100643000010.tif36170

[0419] In one aspect, the linker L1 forms a disulfide bond with the antibody and the linker has the structure: TIFF2023100643000011.tif20170In formula, R 1 , R 2 , R 3 , and R 4 is independently H, an optionally substituted branched or linear C 1 ~C 5 alkyl, and optionally substituted C 3 a carbon atom selected from or bound to the group consisting of ~C6 cycloalkyl and R 1 and R 2 together or R 3 and R 4 together, C 3 ~C 6 Forms a cycloalkyl ring.

[0420] In one aspect, the carbonyl group of the linker is connected to an amine group within the CIDE. Also note that the sulfur atom attached to Ab is the sulfur group from the cysteine ​​in the antibody. In another embodiment, L1 has a functionality that is capable of reacting with free cysteines present on the antibody to form a covalent bond. Non-limiting examples of such reactive functional groups include activated esters such as maleimides, haloacetamides, α-haloacetyls, succinimide esters, 4-nitrophenyl esters, pentafluorophenyl esters, tetrafluorophenyl esters, anhydrides, acid chlorides, sulfonyl chlorides, isocyanates, and isothiocyanates. See, for example, Klussman et al. (2004) Bioconjugate Chemistry 15(4):765-773, conjugation methods at page 766, and the examples therein.

[0421] In some embodiments, the linker has functionalities that are capable of reacting with electrophilic groups present on the antibody. Examples of such electrophilic groups include, but are not limited to, aldehyde and ketone carbonyl groups. In some embodiments, the heteroatom of the reactive functional group of the linker can react with electrophilic groups on the antibody to form a covalent bond with the antibody unit. Non-limiting examples of such reactive functional groups include, but are not limited to, hydrazides, oximes, amino, hydrazines, thiosemicarbazones, hydrazinecarboxylates, and arylhydrazides.

[0422] A linker may comprise one or more linker components. Exemplary linker components include 6-maleimidocaproyl (“MC”), maleimidopropanoyl (“MP”), valine-citrulline (“val-cit” or “vc”), alanine-phenylalanine (“ala-phe”), p-aminobenzyloxycarbonyl (“PAB”), N-succinimidyl 4-(2-pyridylthio)valerate (“SPP”), and 4-(maleimidomethyl)cyclohexane-1 rate (“MCC”). Various linker components are known in the art, some of which are described below.

[0423] The linker may be a "cleavable linker" that facilitates release of CIDE. Non-limiting exemplary cleavable linkers include acid-labile linkers (e.g., including hydrazones), protease-sensitive (e.g., peptidase-sensitive) linkers, photolabile linkers, or disulfide-containing linkers (Chari et al., Cancer Research 52:127-131 (1992), US Pat. No. 5,208,020).

[0424] In certain embodiments, the linker has the formula: TIFF2023100643000012.tif11170 where A is a "stretcher unit" and a is an integer from 0 to 1; W is an "amino acid unit" and w is an integer from 0 to 12; Y is a "spacer unit" and y is 0, 1 or 2. have Exemplary embodiments of such linkers are described in US Pat. No. 7,498,298.

[0425] In some embodiments, the linker moiety comprises a "stretcher unit" that connects the antibody to another linker moiety or CIDE moiety. An exemplary Stretcher unit is shown below (wavy lines indicate sites of covalent attachment to antibodies, CIDE, or additional linker moieties). TIFF2023100643000013.tif115170

[0426] In certain embodiments, the linker is: TIFF2023100643000014.tif54170

[0427] In certain embodiments, the linker has the formula: -A a -Y y - wherein A and Y are defined as above. In certain embodiments the spacer unit Y may be a phosphate such as a monophosphate or a bisphosphate. In certain embodiments, stretcher component A comprises: TIFF2023100643000015.tif28170

[0428] In certain embodiments, the linker is: TIFF2023100643000016.tif29170

[0429] 3. CIDE ("D") Useful CIDEs have the general formula given above. CIDE includes those with the following components:

[0430] a E3 ubiquitin ligase linking group (E3LB) E3 ubiquitin ligases (more than 600 of which are known in humans) confer substrate specificity for ubiquitination. There are known ligands that bind to these ligases. The E3 ubiquitin ligase binding groups described herein are peptides or small molecules capable of binding an E3 ubiquitin ligase selected from the group consisting of von Hippel-Lindau (VHL) and XIAP.

[0431] A specific E3 ubiquitin ligase is the von Hippel-Gendau (VHL) tumor suppressor, a substrate recognition subunit of the E3 ligase complex VCB, composed of elongins B and C, Cul2 and Rbxl. A major substrate of VHL is hypoxia-inducible factor Iα (HIF-Iα), a transcription factor that upregulates genes such as the pro-angiogenic growth factor VEGF and the erythrocyte-inducing cytokine erythropoietin in response to low oxygen levels. Compounds that bind to VHL include hydroxyproline compounds such as those disclosed in WO2013 / 106643, WO2013 / 106646, and others described in U.S. Patent Application Publication No. 2016 / 0045607, WO2014187777, U.S. Patent Application Publication No. 20140356322 and U.S. Patent No. 9,249,153. can be a compound.

[0432] In one aspect, the subject matter herein has the following chemical structure: TIFF2023100643000017.tif49170 In the formula, R. 1’ is C 1 ~C 6 Alkyl groups, optionally substituted -(CH 2 ) n OH, optionally substituted -(CH 2 ) n SH, optionally substituted (OH 2 ) n -O-(C 1 ~C 6 ) alkyl groups, optionally substituted (CH 2 ) n -WCOCW-(C 0 ~C 6 ) alkyl group and each W is independently H or C 1 ~C 3 Alkyl groups, optionally substituted -(CH 2 ) n COOH, optionally substituted -(CH 2 ) n C(O)-(C 1 ~C 6 alkyl), optionally substituted -(CH 2 ) n NHC(O)-R 1 , optionally substituted -(CH 2 ) n C(O)-NR 1 R. 2 , optionally substituted -(CH 2 ) n OC(O)-NR 1 R. 2 , -(CH 2 O) n H, optionally substituted -(CH 2 ) n OC(O)-(C 1 ~C 6 alkyl), optionally substituted -(CH 2 ) n C(O)-O-(C 1 ~C 6 alkyl), optionally substituted -(CH 2 O) n COOH, optionally substituted -(OCH 2 ) n O-(C 1 ~C 6 alkyl), optionally substituted -(CH 2 ) n C(O)-O-(C 1 ~C 6 alkyl), optionally substituted -(OCH 2 ) n NHC(O)-R 1 , optionally substituted -(CH 2 O) n C(O)-NR 1 R. 2 , -(CH 2 CH 2 O) n H, optionally substituted -(CH 2 CH 2 O) n COOH, optionally substituted -(OCH 2 CH 2 ) n O-(C 1 ~C 6 alkyl), optionally substituted -(CH 2 CH 2 O) n C(O)-(C 1 ~C 6 alkyl), optionally substituted -(OCH 2 CH 2 ) n NHC(O)-R 1 , optionally substituted -(CH 2 CH 2 O) n C(O)-NRiR 2 , with -SO substituted as needed 2 R. s , S(O)R, optionally substituted s , NO 2 , CN or halogen (F, Cl, Br, I, preferably F or Cl); R. 1 and R 2 each independently H or C optionally substituted with 1 or 2 hydroxyl groups or up to 3 halogen groups (preferably fluorine) 1 ~C 6 is an alkyl group; R. s is C 1 ~C 6 an alkyl group, an optionally substituted aryl, heteroaryl or heterocyclic group, or -(CH 2 )NR 1 R. 2 is a group; X and X' are each independently C=O, O=S, -S(O), S(O) 2 (preferably both X and X' are C=O); R. 2’ teeth, -(CH 2 ) n -(C=O) u (NR 1 ) v (SO 2 ) w alkyl group, -(CH 2 ) n -(C=O) u (NR 1 )v(SO 2 ) w NR 1N R. 2N group, -(CH 2 )n -(C=O) u (NR 1 ) v (SO 2 ) w -aryl, -(CH 2 ) n -(C=O) u (NR 1 ) v (SO 2 ) w -heteroaryl, -(CH 2 ) n -(C=O) v NR 1 (SO 2 ) w -heterocycle, -NR substituted as needed Z -(CH 2 ) n -C(O) u (NR 1 ) v (SO 2 ) w -alkyl, -NR substituted as needed Z -(CH 2 ) n -C(O) u (NR 1 ) v (SO 2 ) w -NR 1N R. 2N , -NR substituted as needed Z -(CH 2 ) n -C(O) u (NR 1 ) v (SO 2 ) w -NR 1 C(O)R 1N , -NR substituted as needed Z -(CH 2 ) n -(C=O) u (NR 1 ) v (SO 2 ) w -aryl, -NR substituted as needed Z -(CH 2 ) n -(C=O) u (NR 1 ) v (SO 2 ) w -heteroaryl, -NR substituted as needed Z -(CH 2 )n-(C=O) v NR 1 (SO 2 ) w -heterocycle, -X substituted as needed R2’ -alkyl groups, -X substituted as needed R2’ -aryl groups, -X substituted as needed R2’ -heteroaryl groups, -X substituted as needed R2’ - is a heterocyclic group, R. 3’ teeth, optionally substituted alkyl, -(CH 2 ) n -C(O) u (NR 1 ) v (SO 2 ) w -alkyl, -(CH 2 ) n -C(O) u (NR 1 ) v (SO 2 ) w -NR 1N R. 2N , -(CH 2 ) n -C(O) u (NR 1 ) v (SO 2 ) w -NR 1 C(O)R 1N , -(CH 2 ) n -C(O) u (NR 1 ) v (SO 2 ) w -C(O)NR 1 R. 2 , -(CH 2 ) n -C(O) u (NR 1 ) v (SO 2 ) w -aryl, -(CH 2 ) n -C(O)u(NR 1 ) v (SO 2 ) w -heteroaryl, -(CH 2 ) n -C(O) u (NR 1 ) v (SO 2 ) w -heterocycle, -NR substituted as needed Z -(CH 2 ) n -C(O) u (NR 1 ) v (SO 2 ) w -alkyl, -NR substituted as needed Z -(CH 2 ) n -C(O) u (NR 1 ) v (SO 2 ) w -NR 1N R. 2N , -NR substituted as needed Z -(CH 2 ) n -C(O) u (NR 1 ) v (SO 2 ) w -NR 1 C(O)R 1N , -NR substituted as needed Z -(CH 2 ) n -C(O) u (NR 1 ) v (SO 2 ) w -aryl, -NR substituted as needed Z -(CH 2 ) n -C(O) u (NR 1 ) v (SO 2 ) w -heteroaryl, -NR substituted as needed Z -(CH 2 ) n -C(O) u (NR 1 ) v (SO 2 ) w -heterocycle, optionally substituted -O-(CH 2 )n-(C=O) u (NR 1 ) v (SO 2 ) w -alkyl, optionally substituted -O-(CH 2 )n-(C=O) u (NR 1 ) v (SO 2 ) w -NR 1N R. 2N , optionally substituted -O-(CH 2 )n-(C=O) u (NR 1 ) v (SO 2 ) w -NR 1 C(O)R 1N , optionally substituted -O-(CH 2 )n-(C=O) u (NR 1 ) v (SO 2 ) w -aryl, optionally substituted -O-(CH 2 ) n -(C=O) u (NR 1 ) v (SO 2 ) w -heteroaryl, optionally substituted -O-(CH 2 ) n -(C=O) u (NR 1 ) v (SO 2 ) w -heterocycle, -(CH2 ) n -(V) n’ -(CH 2 ) n -(V) n’ -alkyl groups, -(CH 2 ) n -(V) n’ -(CH 2 ) n -(V) n’ -aryl groups, -(CH 2 ) n -(V) n’ -(CH 2 ) n -(V) n’ -heteroaryl groups, -(CH 2 ) n -(V) n’ -(CH 2 ) n -(V) n’ -heterocyclic groups, -(CH 2 ) n -N(R 1’ )(C=O) m’ -(V) n’ -alkyl groups, -(CH 2 ) n -N(R 1’ )(C=O) m’ -(V) n’ -aryl groups, -(CH 2 ) n -N(R 1’ )(C=O) m’ -(V) n’ -heteroaryl groups, -(CH 2 ) n -N(R 1’ )(C=O) m’ -(V) n’ -heterocyclic groups, -X substituted as needed R3’ -alkyl groups, -X substituted as needed R3’ -aryl groups, -X substituted as needed R3’ -heteroaryl groups, -X substituted as needed R3’ - is a heterocyclic group, replaced where necessary; R. 1N and R 2N each independently H, C optionally substituted with 1 or 2 hydroxyl groups and up to 3 halogen groups 1 ~C 6 alkyl, or optionally substituted -(CH 2 ) n -aryl, -(CH 2 ) n -heteroaryl or -(CH 2 ) n - is a heterocyclic group; R. Z and R 1 are each independently H or C 1 ~C 3 is an alkyl group; V is O, S or NR 1 is; R. 1 is the same as above; X R2’ and X R3’ are each independently an optionally substituted -CH 2 ) n -, -CH 2 ) n - CH(X V )=CH(X V )-(cis or trans), -CH 2 ) n -CH≡CH-, -(CH 2 CH 2 O) n -or C 3 ~C 6 is a cycloalkyl group and X v is H, halo, or optionally substituted C 1 ~C 3 is an alkyl group; each m is independently 0, 1, 2, 3, 4, 5, 6; each m' is independently 0 or 1; each n is independently 0, 1, 2, 3, 4, 5, 6; each n' is independently 0 or 1; each u is independently 0 or 1; each v is independently 0 or 1; each w is independently 0 or 1; or a pharmaceutically acceptable salt, enantiomer, diastereomer, solvate or polymorph thereof.

[0433] In an alternative aspect, the invention provides the following chemical structure: TIFF2023100643000018.tif61170 formula, R 1’ , R 2’ and R 3’ are the same as above, and X is a C=O, C=S, -S(O) group or S(O) 2 a group, more preferably a C=O group, or a pharmaceutically acceptable salt, enantiomer, diastereomer, solvate or polymorph thereof.

[0434] In a further preferred embodiment of the invention, a compound according to the invention: TIFF2023100643000019.tif60170 formula, R 1’ , R 2’ and R 3’ is the same as presented above, or a pharmaceutically acceptable enantiomer, diastereomer, solvate or polymorph thereof.

[0435] In a further preferred embodiment of the invention, R 1’ is preferably a hydroxyl group or a group metabolizable to a hydroxyl or carboxylic acid group, preferably a hydroxyl group, so that the compound represents a prodrug form of the active compound. Exemplary Preferred R 1’ Examples of groups include -(CH 2 ) n OH, (CH 2 ) n -O-(C 1 ~C 6 ) alkyl group, -(CH 2 ) n COOH, -(CH 2 O) n H, optionally substituted -(CH 2 ) n C(O)(C 0 ~C 6 )alkyl, optionally substituted -(CH 2 ) n OC(O)-(C 1 C. 6 )alkyl, or optionally substituted -(CH 2 ) n C(O)-O-(C 1 ~C 6 ) alkyl, where n is 0 or 1. Most of the time R 1 is hydroxyl.

[0436] X and X', if present, are preferably C=O, C=S, -S(O) groups or S(O) 2 group, more preferably a C=O group.

[0437] R2' is preferably optionally substituted -NR 1 -T-aryl, optionally substituted -NR 1 -T-heteroaryl group or optionally substituted -NR 1 -T-heterocycle and R 1 is C 1 ~C 3 an alkyl group, preferably H or CH 3 , more preferably H, and T is optionally substituted -(CH 2 ) n - group and each methylene group in the alkylene chain may optionally be substituted with 1 or 2 substituents, preferably halogen, C 1 ~C 3 Alkyl groups or side chains of amino acids described elsewhere herein, preferably 1 or 2 methyl groups, which may be optionally substituted; and n is 0-6, often 0, 1, 2 or 3, preferably 0 or 1. Alternatively, T can also be -(CH 2 O) n - group, -(OCH 2 ) n - group, -(CH 2 CH 2 O) n - group, -(OCH 2 CH 2 ) n - groups, and any of these groups are optionally substituted.

[0438] R. 2’ Preferred aryl groups for include optionally substituted phenyl or naphthyl groups, preferably phenyl groups, where phenyl groups are halogen (preferably F or C1), amine, monoalkyl- or dialkylamine (preferably dimethylamine), F, C1, OH, SH, COOH, C 1 ~C6 alkyl, preferably CH 3 , CF 3 , OMe, OCF 3 , NO 2 , or a CN group (each of which may be substituted at the ortho, meta and / or para, preferably para, positions of the phenyl ring), an optionally substituted phenyl group (the phenyl group itself is preferably F, C, OH, SH, COOH, CH 3 , CF 3 , OMe, OCF 3 , NO 2 or at least one of the CN groups, which may be substituted at the ortho, meta and / or para positions of the phenyl ring, preferably at the para position), optionally substituted naphthyl groups, optionally substituted heteroaryl, optionally substituted isoxazoles, including preferably methyl-substituted isoxazoles, optionally substituted oxazoles, including methyl-substituted oxazoles, optionally substituted thiazoles, including methyl-substituted thiazoles optionally substituted isothiazoles, including methyl-substituted isothiazoles; optionally substituted pyrroles, including methyl-substituted pyrroles; optionally substituted imidazoles, including methylimidazole; optionally substituted benzimidazoles or methoxybenzylimidazoles; optionally substituted triazole group, optionally substituted pyridine group including halo-(preferably F) or methyl substituted pyridine group or oxapyridine group (pyridine group is attached to phenyl group by oxygen), optionally substituted furan, optionally substituted benzofuran, optionally substituted dihydrobenzofuran, optionally substituted indole, indolizine or azaindolizine (2, 3, or 4-azaindolizine) , an optionally substituted quinoline, which may be substituted with optionally substituted groups according to the following chemical structures: TIFF2023100643000020.tif94170S c is CHR SS , NR URE , or O; R. HET is H, CN, NO 2 , halo (preferably C1 or F), optionally substituted C 1 ~C 6 Alkyl (preferably 1 or 2 hydroxyl groups, or up to 3 halo groups (e.g. CF 3 ), optionally substituted O(C 1 ~C 6 alkyl) (preferably substituted with 1 or 2 hydroxyl groups, or up to 3 halo groups), or an optionally substituted acetylene group -C≡C-R a and R a is H or C 1 ~C 6 Alkyl group (preferably C 1 ~C3 alkyl); R. SS is H, CN, NO 2 , halo (preferably F or C1), optionally substituted C 1 ~C 6 Alkyl (preferably substituted with 1 or 2 hydroxyl groups, or up to 3 halo groups), optionally substituted O(C 1...

Claims

[Claim 1] An invention described in the specification.

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