Companion diagnostics for therapeutic agents directed against human CEACAM1

Abstract

Provided herein are methods and compositions for determining whether to administer a CEACAM1 antibody or antigen-binding agent as a treatment to a subject diagnosed with a CEACAM1-associated disease or disorder. TIFF2025506405000027.tif68128

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit under 35 USC § 119(e) of U.S. Provisional Application No. 63 / 306,147, filed February 3, 2022, the contents of which are incorporated by reference herein in their entirety.

[0002] Government support This invention was made with Government support under Grant No. DK51362 awarded by the National Institutes of Health. The Government has certain rights in this invention.

[0003] Sequence Listing This application contains a Sequence Listing that has been submitted through the Patent Center in XML format, which is incorporated herein by reference in its entirety. The XML copy was created on February 2, 2023, is entitled "043214-191430USPX_SL.xml", and is 23,259 bytes in size.

[0004] Technical Field The technology described herein relates to the fields of molecular biology and medicine. More specifically, the technology relates to determining suitable treatments for treating subjects with CEACAM1-related diseases or disorders. [Background technology]

[0005] background Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) is a member of the carcinoembryonic antigen (CEA) family of immunoglobulin (Ig)-like transmembrane glycoproteins. CEACAM family members are involved in cell-cell recognition and regulate cellular processes ranging from shaping tissue architecture and angiogenesis to regulation of insulin homeostasis and T-cell proliferation. The CEACAM family includes CEACAM1, CEACAM3, CEACAM5, CEACAM6, CEACAM7, CEACAM8, CEACAM16, CEACAM18, CEACAM19, CEACAM20, and CEACAM21.

[0006] The immune system protects the body from foreign invaders and diseased cells, but immune disorders, specifically those associated with T-cell tolerance, such as cancer, can cause great damage. According to the most recent data from the World Health Organization, 10 million people were diagnosed with cancer worldwide in the year 2000, resulting in 6 million deaths from it. Furthermore, statistics show that the incidence of cancer is rising globally. In the United States, for example, projections suggest that 50 percent of people alive today will be diagnosed with some form of cancer at some point in their lives.

[0007] Modern technologies, such as those involving the use of hybridomas, have made available a source of highly specific and potent monoclonal antibodies that are useful in common diagnostic and clinical procedures. Summary of the Invention

[0008] overview Diseases or disorders in which CEACAM1 plays a role in pathology can be treated with inhibitors of CEACAM1.Antibody or construct, part or fragment thereof, that comprises antigen-binding domain that binds to CEACAM1 can be used as an inhibitor of protein function.On the other hand, allelic differences in CEACAM1 that affect the epitope displayed by a given CEACAM1 polypeptide make some antibodies more effective than others.The technology described herein relates to methods and compositions for determining treatment for subjects diagnosed with CEACAM1-related diseases or disorders by detecting the genotype of the subject and then determining whether to administer an antibody or antigen-binding substance that binds to a given face or domain of CEACAM1.

[0009] In one aspect, a method is described herein that includes receiving the results of an hCEACAM1 genotyping assay performed on a sample from a subject in need of treatment for a CEACAM1-related disease or disorder; and if the subject's hCEACAM1 genotype is homozygous wildtype for A49, Q89, Q44, and Y34, or heterozygous for A49V, Q89H, Q44L, and / or Y34C, selecting a treatment with an agent comprising an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1, and if the subject's hCEACAM1 genotype is homozygous for the A49V, Q89H, Q44L, and / or Y34C hCEACAM1 alleles, selecting a treatment with an agent other than an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1.

[0010] In another aspect, the method includes genotyping a sample from a subject in need of treatment for a CEACAM1-related disease or disorder to detect the hCEACAM1 genotype of the subject; and if the subject's hCEACAM1 genotype is homozygous wildtype for A49, Q89, Q44, and Y34, or heterozygous for A49V, Q89H, Q44L, and / or Y34C, selecting treatment with an agent comprising an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1, and if the subject's hCEACAM1 genotype is homozygous for the A49V, Q89H, Q44L, and / or Y34C hCEACAM1 alleles, selecting treatment with an agent other than an antibody or agent comprising an antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1.

[0011] In one embodiment of this aspect and all other aspects provided herein, the subject has or has been diagnosed with a chronic disease.

[0012] In one embodiment of this aspect and all other aspects provided herein, the chronic disease is selected from the group consisting of cancer; persistent infection; and chronic viral infection.

[0013] In one embodiment of this aspect and all other aspects provided herein, the subject is a human.

[0014] In one embodiment of this aspect and all other aspects provided herein, an antibody or antigen binding domain that binds to an epitope of the GF-CC' domain of CEACAM1 comprises a heavy chain variable region and a light chain variable region; the heavy chain variable region and the light chain variable region each comprise CDR1, CDR2, and CDR3; the sequence of CDR1H comprises the sequence X1HX2X3S (SEQ ID NO: 1); Xi is A, D, N, or S; X2 is A or G; X3 is an amino acid having a hydrophobic side chain, including I or M; and the sequence of CDR2H is TIFF2025506405000002.tif4128; the sequence of CDR3H is X4 is D, G, or P; X5 is F or P; Xe is D or F; X7 is A or Y; Xs is L, H, or F; and the sequence of CDR1L is the sequence of CDR2L is LTSNRAT (SEQ ID NO:5); the sequence of CDR3L is X9 is W or N; X10 is S or T; Xu is A or an amino acid having a neutral hydrophilic side chain, including S, N, and T; X12 is L, F, or N; and X13 is P or F.

[0015] In one embodiment of this aspect and all other aspects provided herein, the sample is selected from a blood sample, a tissue sample, a tumor sample, and a cell sample.

[0016] In one embodiment of this aspect and all other aspects provided herein, the genotyping assays include direct probing, allele-specific hybridization, PCR, allele-specific amplification (ASA), single base extension, ARMS-PCR, Taqman, oligoligation assays, single-stranded conformation analysis (SSCP), Genetic Bit Analysis and RFLP direct sequencing, mass spectrometry (MALDI-TOF), and DNA arrays.

[0017] In one embodiment of this aspect and all other aspects provided herein, the method further comprises administering a selected therapy.

[0018] In one embodiment of this aspect and all other aspects provided herein, the method further includes, if the CEACAM1 genotype is heterozygous for A49V, Q89H, Q44L, and / or Y34C, assaying a sample from the subject containing CEACAM1 to determine the extent to which the subject's CEACAM1 is bound by an agent comprising an antibody or antigen binding domain that binds to an epitope in the GF-CC' domain of CEACAM1, and adjusting the amount and / or dosage of the agent to be administered as a function of the degree of binding of the agent to the subject's CEACAM1.

[0019] In one embodiment of this aspect and all other aspects provided herein, the assaying step comprises a method selected from flow cytometry, mass cytometry, immunohistochemistry, immunofluorescence or FRET assay, surface plasmon resonance, radioimmunoassay, and biolayer interferometry.

[0020] In one embodiment of this aspect and all other aspects provided herein, the method further comprises extracting genomic DNA from the sample obtained from the subject prior to the genotyping step.

[0021] In another aspect, described herein is a method of treating a subject in need of treatment for a CEACAM1-associated disease or disorder, the method comprising: genotyping a sample from the subject to detect the subject's hCEACAM1 genotype; and administering a therapeutically effective amount of an agent comprising an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1 if the subject's hCEACAM1 genotype is homozygous wild type for A49, Q89, Q44, and Y34; and administering a therapeutically effective amount of an agent comprising an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1 if the subject's hCEACAM1 genotype is heterozygous for A49V, Q89H, Q44L, and / or Y34C. The method includes assaying a sample from the subject containing CEACAM1 to determine the extent to which the subject's CEACAM1 is bound by an agent containing an antibody or antigen-binding domain, and administering a therapeutically effective amount of the agent, wherein the amount or dosing schedule of the agent administered is adjusted as a function of the degree of binding of the agent to the subject's CEACAM1; and if the subject's hCEACAM1 genotype is homozygous for A49V, Q89H, Q44L, and / or Y34C, administering a therapeutically effective amount of treatment with an antibody or agent other than one containing an antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1, thereby treating the CEACAM1-related disease or disorder.

[0022] In one embodiment of this aspect and all other aspects provided herein, the subject has or has been diagnosed with a chronic disease.

[0023] In one embodiment of this aspect and all other aspects provided herein, the chronic disease is selected from the group consisting of cancer; persistent infection; and chronic viral infection.

[0024] In one embodiment of this aspect and all other aspects provided herein, the subject is a human.

[0025] In one embodiment of this aspect and all other aspects provided herein, an antibody or antigen binding domain that binds to an epitope of the GF-CC' domain of CEACAM1 comprises a heavy chain variable region and a light chain variable region; the heavy chain variable region and the light chain variable region each comprise CDR1, CDR2, and CDR3; the sequence of CDR1H comprises the sequence X1HX2X3S (SEQ ID NO:1); Xi is A, D, N, or S; X2 is A or G; X3 is an amino acid having a hydrophobic side chain, including I or M; and the sequence of CDR2H is TIFF2025506405000006.tif4128; the sequence of CDR3H is X4 is D, G, or P; X5 is F or P; Xe is D or F; X7 is A or Y; Xs is L, H, or F; the sequence of CDR1L comprises the sequence RANSAVSYMY (SEQ ID NO:4); the sequence of CDR2L comprises the sequence LTSNRAT (SEQ ID NO:5); the sequence of CDR3L comprises the sequence X9 is W or N; X10 is S or T; Xu is A or an amino acid having a neutral hydrophilic side chain, including S, N, and T; X12 is L, F, or N; and X13 is P or F.

[0026] In one embodiment of this aspect and all other aspects provided herein, the sample is selected from a blood sample, a tissue sample, a tumor sample, and a cell sample.

[0027] In one embodiment of this aspect and all other aspects provided herein, the genotyping assays include direct probing, allele-specific hybridization, PCR, allele-specific amplification (ASA), single base extension, ARMS-PCR, Taqman, oligoligation assays, single-stranded conformation analysis (SSCP), gene bit analysis and RFLP direct sequencing, mass spectrometry (MALDI-TOF), and DNA arrays.

[0028] In one embodiment of this aspect and all other aspects provided herein, the step of assaying a sample from a subject containing CEACAM1 to determine the extent to which the subject's CEACAM1 is bound by an agent includes a method selected from flow cytometry, mass cytometry, immunohistochemistry, immunofluorescence or FRET assay, surface plasmon resonance, radioimmunoassay, and biolayer interferometry.

[0029] In another aspect, an assay is described herein that includes the steps of: (i) extracting genomic DNA from a biological sample from a subject in need of treatment for a CEACAM1-associated disease or disorder; (ii) genotyping the subject's DNA for sequences corresponding to CEACAM A49, Q89, Q44, and Y34, thereby determining whether the subject is homozygous wild type for A49, Q89, Q44, and Y34, or homozygous or heterozygous for each of A49V, Q89H, Q44L, and Y34C.

[0030] In one embodiment of this aspect and all other aspects provided herein, the CEACAM1-associated disease or disorder is selected from the group consisting of cancer; persistent infection; and chronic viral infection.

[0031] In one embodiment of this aspect and all other aspects provided herein, the subject is a human.

[0032] In one embodiment of this aspect and all other aspects provided herein, the biological sample is selected from a blood sample, a tissue sample, a tumor sample, and a cell sample.

[0033] In one embodiment of this aspect and all other aspects provided herein, the genotyping step comprises direct probing, allele-specific hybridization, PCR, allele-specific amplification (ASA), single base extension, ARMS-PCR, Taqman, oligoligation assay, single-stranded conformation analysis (SSCP), gene bit analysis and RFLP direct sequencing, mass spectrometry (MALDI-TOF), and DNA arrays.

[0034] In one embodiment of this aspect and all other aspects provided herein, the assay further includes (iii) assaying the subject's CEACAM1 protein to determine the extent to which the subject's CEACAM1 is bound by an antibody or antigen binding domain that binds to an epitope in the GF-CC' domain of CEACAM1.

[0035] In one embodiment of this aspect and all other aspects provided herein, the assaying step of step (iii) comprises a method selected from flow cytometry, mass cytometry, immunohistochemistry, immunofluorescence or FRET assay, surface plasmon resonance, radioimmunoassay, and biolayer interferometry.

[0036] In another aspect, an assay is described herein that includes (i) extracting a protein comprising CEACAM1 from a biological sample from a subject in need of treatment for a CEACAM1-associated disease or disorder, and (ii) assaying the subject's CEACAM1 in the extracted protein to determine the extent to which the subject's CEACAM1 is bound by an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1.

[0037] In one embodiment of this aspect and all other aspects provided herein, the assaying step of step (ii) comprises a method selected from flow cytometry, mass cytometry, immunohistochemistry, immunofluorescence or FRET assay, surface plasmon resonance, radioimmunoassay, and biolayer interferometry.

[0038] In one embodiment of this aspect and all other aspects provided herein, the assay further comprises (iii) genotyping the subject's DNA for sequences corresponding to CEACAM A49, Q89, Q44, and Y34, thereby determining whether the subject is homozygous wild type for A49, Q89, Q44, and Y34, or homozygous or heterozygous for each of A49V, Q89H, Q44L, and Y34C.

[0039] In one embodiment of this aspect and all other aspects provided herein, the genotyping step comprises direct probing, allele-specific hybridization, PCR, allele-specific amplification (ASA), single base extension, ARMS-PCR, Taqman, oligoligation assay, single-stranded conformation analysis (SSCP), gene bit analysis and RFLP direct sequencing, mass spectrometry (MALDI-TOF), and DNA arrays.

[0040] In another aspect, the method includes analyzing genomic DNA from a biological sample obtained from the subject for the subject's genotype for alleles of hCEACAM1 at sites corresponding to A49, Q89, Q44, and Y34, and, if the genotype is found to be homozygous for A49, Q89, Q44, and Y34, or heterozygous for one or more of the A49V, Q89H, Q44L, and Y34C alleles of CEACAM1, detecting a genomic DNA fragment that binds to an epitope in the GF-CC' domain of CEACAM1. Described herein are methods for selecting an hCEACAM1 antibody therapy for a subject in need thereof, comprising selecting treatment with an agent comprising an hCEACAM1 antibody or antigen-binding fragment thereof that binds to an epitope in the GF-CC' domain of CEACAM1, or, if the genotype is found to be homozygous for one or more of the A49V, Q89H, Q44L, and Y34C alleles of CEACAM1, selecting treatment with an agent other than an hCEACAM1 antibody or antigen-binding fragment thereof that binds to an epitope in the GF-CC' domain of CEACAM1.

[0041] In one embodiment of this aspect and all other aspects provided herein, the subject has or has been diagnosed with a chronic disease.

[0042] In one embodiment of this aspect and all other aspects provided herein, the chronic disease is selected from the group consisting of cancer; persistent infection; and chronic viral infection.

[0043] In one embodiment of this aspect and all other aspects provided herein, the subject is a human.

[0044] In one embodiment of this aspect and all other aspects provided herein, an antibody or antigen binding domain that binds to an epitope of the GF-CC' domain of CEACAM1 comprises a heavy chain variable region and a light chain variable region; the heavy chain variable region and the light chain variable region each comprise CDR1, CDR2, and CDR3; the sequence of CDR1H comprises the sequence X1HX2X3S (SEQ ID NO:1); X1 is A, D, N, or S; X2 is A or G; and X3 is an amino acid having a hydrophobic side chain, including I or M; and the sequence of CDR2H is TIFF2025506405000009.tif4128; the sequence of CDR3H is X4 is D, G, or P; X5 is F or P; X6 is D or F; X7 is A or Y; and X8 is L, H, or F; the sequence of CDR1L comprises the sequence RANSAVSYMY (SEQ ID NO:4); the sequence of CDR2L comprises the sequence LTSNRAT (SEQ ID NO:5); the sequence of CDR3L comprises the sequence TIFF2025506405000011.tif4128; X9 is W or N; X 10 is S or T;X 11 is A or an amino acid having a neutral hydrophilic side chain, including S, N, and T; X 12 is L, F, or N;X 13 is P or F.

[0045] In one embodiment of this aspect and all other aspects provided herein, the sample is selected from a blood sample, a tissue sample, a tumor sample, and a cell sample.

[0046] In one embodiment of this aspect and all other aspects provided herein, genotyping includes direct probing, allele-specific hybridization, PCR, allele-specific amplification (ASA), single base extension, ARMS-PCR, Taqman, oligoligation assay, single-stranded conformation analysis (SSCP), gene bit analysis and RFLP direct sequencing, mass spectrometry (MALDI-TOF), and DNA arrays.

[0047] In one embodiment of this aspect and all other aspects provided herein, the method further comprises administering a selected therapy.

[0048] In one embodiment of this aspect and all other aspects provided herein, the method further includes, if the CEACAM1 genotype is heterozygous for A49V, Q89H, Q44L, and / or Y34C, assaying a sample from the subject containing CEACAM1 to determine the extent to which the subject's CEACAM1 is bound by an agent comprising an antibody or antigen binding domain that binds to an epitope in the GF-CC' domain of CEACAM1, and adjusting the amount and / or dosage of the agent to be administered as a function of the degree of binding of the agent to the subject's CEACAM1.

[0049] In one embodiment of this aspect and all other aspects provided herein, the assaying step comprises a method selected from flow cytometry, mass cytometry, immunohistochemistry, immunofluorescence or FRET assay, surface plasmon resonance, radioimmunoassay, and biolayer interferometry.

[0050] definition For convenience, certain terms used in the specification, examples, and the appended claims are collected here. Unless otherwise stated or implied from the context, the following terms and phrases include the meanings provided below. Unless otherwise stated or clear from the context, the following terms and phrases do not exclude the meaning that the term or phrase has acquired in the technical field to which it belongs. The definitions are provided to help explain certain embodiments, and are not intended to limit the claimed invention, since the scope of the invention is limited only by the claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the invention belongs.

[0051] As used herein, "reducing pathology" refers to a reduction in one or more symptoms or markers of a CEACAM1-associated disease or disorder by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more.

[0052] As used herein, "CEACAM1-associated disease or disorder" refers to a disease or disorder in which CEACAM1-mediated signal transduction contributes to the disease pathology (e.g., CEACAM1 expression in lesions such as cancer and tumor cells themselves), and the disease or disorder can be effectively treated by inhibiting CEACAM1 function, including inhibiting the interaction of CEACAM1 with one or more CEACAM1 ligands.Symptoms of CEACAM1-associated disease or disorder include changes in the infiltration pattern of immune cells in the microenvironment, refractoriness of T cells, low levels of T cells, or elevated levels or increases in regulatory T cells relative to appropriate reference or control.

[0053] The terms "reduce", "reduced", "reduction", or "inhibit" are all used herein to mean a statistically significant reduction or alleviation of a characteristic, level, or other parameter (such as the amount or relative level of a disease symptom). In some embodiments, "reduce", "reduction", or "reduce" or "inhibit" typically means a reduction of at least 10% of a characteristic, level, or other parameter compared to a reference, and may include, for example, a reduction of at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or more. As used herein, "reduce" or "inhibit" does not encompass a complete inhibition or reduction compared to a reference level. Rather, the term "complete inhibition" is used to refer to 100% inhibition when compared to an appropriate reference level. The reduction in a given cancer symptom may preferably be a decrease to a level that is accepted as being within the normal range for individuals without cancer.

[0054] The terms "increased," "increase," or "enhance," or "activate" are all used herein to generally mean an increase in a characteristic, level, or other parameter (e.g., the amount or relative level of a disease parameter, marker, or symptom) by a statistically significant amount. For the avoidance of any doubt, the terms "increased," "increase," or "enhance," or "activate" refer to an increase in the level of a parameter, marker, or symptom by at least 10% compared to a reference level (e.g., the level in one or more healthy individuals), for example, an increase of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90%, or up to and including a 100% increase compared to the reference level, or any increase between 10-100%, or at least about 2-fold, or at least about 3-fold, or at least about 4-fold, or at least about 5-fold, or at least about 10-fold, at least about 20-fold, at least about 50-fold, at least about 100-fold, at least about 1000-fold, or more.

[0055] As used herein, an "antibody agent" is a therapeutic composition comprising an anti-CEACAM1 antibody or an antigen-binding portion or fragment thereof. The antibody or antigen-binding portion or fragment thereof in the antibody agent may be configured in any of a number of formats, including, but not limited to, a natural antibody, a humanized or chimeric antibody, a scFv, a bispecific or multispecific antibody, a dAb, a Fab, a Fab', a F(ab')2, a Fd, a Fd', a diabody, or a linear antibody, among others.

[0056] As used herein, a "CP08-based antibody agent" includes an antibody agent whose antigen-binding domain has a heavy chain variable region and a light chain variable region, each of which comprises CDR1, CDR2, and CDR3; the sequence of CDR1H comprises the sequence X1HX2X3S (SEQ ID NO:1); X1 is A, D, N, or S; X2 is A or G; X3 is an amino acid having a hydrophobic side chain, including I or M; and the sequence of CDR2H is TIFF2025506405000012.tif4128; the sequence of CDR3H is X4 is D, G, or P; X5 is F or P; X6 is D or F; X7 is A or Y; and X8 is L, H, or F; the sequence of CDR1L comprises the sequence RANSAVSYMY (SEQ ID NO:4); the sequence of CDR2L comprises the sequence LTSNRAT (SEQ ID NO:5); the sequence of CDR3L comprises the sequence TIFF2025506405000014.tif4128; X9 is W or N; X 10 is S or T;X 11 is A or an amino acid having a neutral hydrophilic side chain, including S, N, and T; X 12 is L, F, or N;X 13 is P or F. In one embodiment, the CP08-based antibody agent has the CDRs of SEQ ID No 1, 2, 3, 4, 5, 6 or affinity matured versions thereof.

[0057] As used herein, the terms "comprising" or "comprises" are used in reference to compositions, methods, and their respective component(s) that are essential to the invention, but leave room for the inclusion of unspecified elements, whether or not essential.

[0058] As used herein, the term "consisting essentially of" refers to an element that is required for a given embodiment. The term permits the presence of additional elements that do not materially affect the basic and novel or functional characteristic(s) of that embodiment of the invention.

[0059] The term "consisting of" refers to the compositions, methods, and their respective components described herein excluding any element not recited in that description of an embodiment.

[0060] As used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "the method" includes one or more methods and / or steps of the type described herein and / or that will become apparent to those of skill in the art upon reading this disclosure. It will be understood that the foregoing detailed description and the examples set forth below are illustrative only and should not be construed as limitations on the scope of the invention. Various changes and modifications to the aspects of the disclosure that will become apparent to those of skill in the art can be made without departing from the spirit and scope of the invention. In addition, all identified patents, patent applications, and publications are expressly incorporated herein by reference for the purpose of describing and disclosing, for example, the methodologies described in such publications that may be used in connection with the present invention. Such publications are provided solely for their disclosure prior to the filing date of this application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by reason of prior invention or for any other reason. All statements as to the date or presentation of the contents of such documents are based on the information available to the applicants and do not constitute any admission as to the accuracy of the dates or contents of such documents. [Brief description of the drawings]

[0061] [Figure 1] Figure 1 shows the crystal structure depicting the CEACAM1:CEACAM1 homodimer interface (PDB ID: 4QXW). One CEACAM1 monomer is shown on the left and the other on the right. Residues Y34, Q44, Q89, and N97 form the YQQN pocket. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0062] Detailed Description Allelic differences in CEACAM1 polypeptide make some CEACAM1-related diseases less amenable to treatment with certain CEACAM1 inhibitors. The technology described herein relates to a method for determining whether a given subject diagnosed with a CEACAM1-related disease or disorder is a candidate for treatment with a given anti-CEACAM1 inhibitor. Specifically, allelic differences affecting the surface of CEACAM1 known as the GF-CC' face (also referred to as the FG-CC' face) affect whether an antibody that interacts with this face is therapeutically effective. The method described herein generally relates to a method for determining a subject's genotype for CEACAM1, which genotype provides information on whether an antibody or antigen-binding agent that interacts with the GF-CC' face is likely to be effective in treating the subject's CEACAM1-related disease or disorder. The decision of whether to administer a given antibody or antigen-binding substance that interacts with the GF-CC' face of CEACAM1, or whether to administer a relatively high or low dose of such a substance, can be informed by genotype, which can be homozygous wild type, heterozygous for one or more allele variants, or homozygous for one or more allele variants that affect the GF-CC' face of CEACAM1.If a subject is homozygous for one or more allele differences that affect the GF-CC' face, an antibody or antigen-binding fragment that binds to the wild type GF-CC' face is unlikely to be effective.On the other hand, if a subject is homozygous wild type or heterozygous for one or more alleles that affect the GF-CC' face, an antibody or antigen-binding fragment that binds to the wild type face is more likely to be effective, for example, at a first dose or frequency for homozygous wild type, or at a second higher dose for heterozygous individuals.

[0063] The following describes considerations and methodologies that will facilitate one of ordinary skill in the art in practicing the techniques disclosed herein.

[0064] CEACAM1 Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) is a human transmembrane glycoprotein and a member of the carcinoembryonic antigen (CEA) gene family, which is a member of the immunoglobulin superfamily. It is also known as cluster of differentiation 66a (CD66a). CEACAM1 family members are involved in cell-cell recognition and regulate cellular processes ranging from tissue architecture shaping and angiogenesis to insulin homeostasis and regulation of T-cell proliferation.

[0065] A variety of cellular activities have been attributed to the CEACAM1 protein, including roles in differentiation and arrangement of tissue three-dimensional structures, angiogenesis, apoptosis, tumor suppression, metastasis, and regulation of innate and adaptive immune responses. Cell types that express CEACAM1 include certain tumor cells, T cells, natural killer (NK) cells, and certain macrophages.

[0066] For example, high CEACAM1 expression occurs in various cancers, such as melanoma, colorectal, gastric, pancreatic, bladder, and thyroid cancers, and is associated with worse tumor progression, metastasis, and poor clinical prognosis. Non-small cell lung cancer (NSCLC), for example, exhibits high microvascular density, distant metastasis, and shorter median overall survival and progression-free survival with high CEACAM1 expression. CEACAM1 expression is also strongly correlated with distant metastasis of pancreatic adenocarcinoma. CEACAM1 expression on tumors promotes CEACAM1-mediated inhibition of T cells and NK cells. As a result, inhibiting CEACAM1 activity can inhibit tumor cell metastasis and the formation of cancer stem cell niches.

[0067] CEACAM1 is also expressed in certain immune system cells and plays a role in immune suppression and immune cell exhaustion. High expression of CEACAM1 on tumor infiltrating lymphocytes (TILs) and other tumor infiltrating immune cells from gastric cancer, lung cancer, melanoma, colorectal cancer and glioma is associated with poor prognosis, for example. With regard to T cells, CEACAM1 expression is nearly eliminated from resting (naive) T cells, while the protein is expressed at high levels on activated T cells. CEACAM1-L is the dominant isoform expressed on most T cells, and acts as an inhibitory receptor that downregulates T cell activation and suppresses T cell function. Therefore, inhibition of CEACAM1 on T cells can restore T cell activity and increase antitumor responses.

[0068] CEACAM1 also mediates cell-cell adhesion through the extracellular portion of CEACAM1, which contains an IgV-like N-domain that is involved in homophilic (CEACAM1:CEACAM1) and heterophilic interactions (e.g., with CEA, CEACAM5, CEACAM8, T-cell immunoglobulin and mucin domain-containing 3 (TIM-3) protein, Helicobacter pylori adhesin HopQ, Neisseria gonorrhoeae / meningitidis opacity protein (OPA), Moraxella sp. Opa-like protein O1pA, Haemophilus influenzae outer membrane protein (OMP) P1, Haemophilus aegyptius OMP P1, Candida albicans, and influenza viruses such as H5N1). TIM-3 was identified as a Th1-specific cell surface protein expressed on activated T cells, dendritic cells, and a subset of macrophages and NK cells. TIM-3 is an activation-induced inhibitory molecule that is involved in tolerance and has been shown to induce T cell exhaustion in chronic viral infections and cancer. CEACAM1, also expressed on activated T cells, has been shown to interact with TIM-3, and this interaction is important for TIM-3-mediated T cell inhibition.

[0069] As shown above, CEACAM1 also serves as a cellular receptor on the apical membrane of mucosal cells for various Gram-negative bacterial pathogens and fungal pathogens such as Candida albicans that are associated with human mucosa. For example, Neisseria gonorrhoeae, Neisseria meningitidis, Streptococcus catarrhalis, Haemophilus influenzae, Haemophilus aegyptius, and pathogenic Escherichia coli strains have well-characterized CEACAM1-binding adhesins. CEACAM1 association with bacterial adhesins triggers endocytosis of bacteria into epithelial cells and transcytosis of microorganisms through intact epithelial layers, thereby allowing microorganisms to exploit CEACAM1 during mucosal colonization. Furthermore, CEACAM1 has been implicated in infections by influenza virus H5N1 and by filamentous nematodes such as Wucheria bancrofti.

[0070] Human CEACAM1 (hCEACAM1) is a single-pass type I transmembrane protein expressed as 12 alternatively spliced ​​isoforms that all contain an N-terminal V-set fold of the immunoglobulin superfamily (IgV) extracellular domain, followed by up to three type 2 constant immunoglobulin (IgC2) extracellular domains (A1, B, A2), a transmembrane sequence, and a signaling cytoplasmic domain. Depending on the splice differential, both cytoplasmic domains contain either a long (L) sequence containing two immunoreceptor tyrosine-based inhibitory motifs (ITIMs) or a short (S) domain lacking ITIMs, which confer intracellular inhibitory or non-inhibitory signals, respectively.

[0071] CEACAM1 function is triggered by cell-cell or trans-binding of the IgV domain, leading to higher-order surface CEACAM1 oligomerization and subsequent intracellular signaling. In contrast to other immune receptors such as T cell inhibitory mucin domain-containing protein 3 (TIM-3), programmed cell death protein 1 (PD-1), and programmed cell death ligand 1 (PD-L1), CEACAM1 serves as its own primary ligand due to the high-affinity homophilic interaction of its unique IgV domain, as well as an important microbial receptor. At basal steady state, CEACAM1 shuttles between monomeric and cis-homodimeric forms on the cell surface.

[0072] The hCEACAM1 IgV domain contains 108 amino acids arranged in nine beta strands (ABCC'C''DEFG) that fold into the conserved IgV antiparallel beta-sandwich tertiary structure used by other IgV-containing immune receptors including TIM-3, PD-1, and PD-L1. The opposing ABED and GFCC' faces of the CEACAM1 beta-sandwich are tethered by an internal salt bridge (R64:D82) that mimics the stabilizing covalent disulfide linkage found in most Ig domains. The more predominant oligomeric form is a high-affinity GFCC'-mediated homodimer that is conserved among other IgV domain-containing proteins. Unique to the hCEACAM1 IgV GFCC' surface is a prominent protrusion of the CC' loop that is strikingly different from the ordered β-hairpins observed in other reported IgV structures. This displaced CC' loop forms a cleft with the FG loop that exposes key residues F29, S32, Y34, V39, G41, Q44, Q89, I91, N97, and E99, which are critical in mediating homophilic CEACAM1 interactions. The structure of hCEACAM1, including the structure of the GF-CC' face, is shown in Huang et al., Nature 517: 386-390 (2015), which is incorporated by reference in its entirety.

[0073] The hCEACAM1 IgV GFCC' face represents the major face involved in homodimer and heterodimer formation. When hCEACAM1 transitions from a dimer to a monomer via disruption of the CC' loop interaction, enhanced thermal motions within the CC' and FG loops result in conformational dynamics that generate monomeric hCEACAM1 species that are more susceptible to both homophilic and heterophilic interactions. Homophilic interactions with neighboring hCEACAM1 monomers, either in cis or trans, mediated by the GFCC' face of the hCEACAM1 monomer, return the destabilized hCEACAM1 monomer to a more thermally stable dimeric form. The GFCC' face stabilized homodimers participate in higher order oligomer formation through minor interactions mediated through ABED. Selective targeting of the GFCC' face of hCEACAM1 with therapeutic agents that exceed the natural homodimer and heterodimer affinity can disrupt the formation of hCEACAM1 homodimers and complexes with other ligands, such as hTIM3, PD-1, and PD-L1, respectively. Additional information regarding the GFCC' surface of CEACAM1 can be found in Huang, Yu-Hwa et al. 2015. CEACAM1 regulates TIM-3-mediated tolerance and exhaustion. Nature 517(7534): 386-390; Gandhi et al. 2021. Structural basis of the dynamic human CEACAM1 monomer-dimer equilibrium. Communications Biology 4, Article number: 360; and Gandhi, AK et al. 2022. Structural analysis of human CEACAM1 oligomerization. Communications Biology 5, Article number 1042; which are incorporated by reference herein in their entireties.

[0074] CEACAM1-Related Disease or Disorder As used herein, a CEACAM1-associated disease or disorder is one in which CEACAM1-mediated signal transduction contributes to disease pathology, and can be effectively treated to reduce such pathology by inhibiting CEACAM1 function, including inhibiting the interaction of CEACAM1 with one or more CEACAM1 ligands. Symptoms or markers of a CEACAM1-associated disease or disorder include changes in the infiltration pattern of immune cells into the microenvironment, T cell refractoriness, low levels of T cells, increased regulatory T cells, and / or increased expression of CEACAM1.

[0075] Examples of CEACAM1-related diseases or disorders include, but are not limited to, cancer or chronic infection.In some embodiments, the subject has cancer.Cancer that migrates from its initial location and takes root in vital organs can eventually lead to the death of the subject through the functional decline of affected organs.Hematopoietic cancers such as leukemia can overwhelm the normal hematopoietic compartment in the subject, thereby causing hematopoietic failure (in the form of anemia, thrombocytopenia, and neutropenia) and ultimately causing death.

[0076] Metastasis is an area of ​​cancer cells that is different from the primary tumor location, resulting from the spread of cancer cells from the primary tumor to other parts of the body. At the time of diagnosis of the primary mass, the subject can be monitored for the presence of metastasis. Metastasis is most often detected through the use of magnetic resonance imaging (MRI) scans, computed tomography (CT) scans, blood and platelet counts, liver function tests, chest X-rays, and bone scans, alone or in combination, in addition to monitoring for certain symptoms.

[0077] Cancers include, but are not limited to, basal cell carcinoma, biliary tract cancer; bladder cancer; bone cancer; brain and CNS cancer; breast cancer; cervical cancer; choriocarcinoma; colon and rectal cancer; connective tissue cancer; cancer of the digestive system; endometrial cancer; esophageal cancer; eye cancer; cancer of the head and neck; gastric cancer; intraepithelial neoplasia; renal cancer; laryngeal cancer; leukemia; liver cancer; lung cancer (e.g., small cell and non-small cell); lymphoma, including Hodgkin's lymphoma and non-Hodgkin's lymphoma; melanoma; myeloma; neuroblastoma; oral cancer (e.g., lip, tongue, mouth, and pharynx); ovarian cancer; pancreatic cancer; prostate cancer; retinoblastoma; rhabdomyosarcoma; rectal cancer; cancer of the respiratory system; sarcoma; skin cancer; stomach cancer; testicular cancer; thyroid cancer; uterine cancer; cancer of the urinary system, and other cancers and sarcomas.

[0078] Response to anticancer treatment refers to any response of the cancer or tumor(s) of the subject to treatment, and preferably refers to the change in tumor amount and / or volume after the start of treatment.For example, tumor response can be evaluated when the initial size and dimensions measured by CT, PET, mammogram, ultrasound, or palpation can be compared with the size of the tumor after intervention, and when the cellularity of the tumor can be estimated histologically and compared with the cellularity of the tumor biopsy taken after the start of treatment.Response can also be evaluated by caliper measurement or pathological examination of the tumor after biopsy or surgical resection. Responses may be recorded in a quantitative manner, such as percent change in tumor volume or cellularity, or using semi-quantitative scoring systems, such as residual cancer burden (Symmans et al., J. Clin. Oncol. (2007) 25:4414-5 4422) or Miller-Payne score (Ogston et al., Breast (Edinburgh, Scotland) (2003) 12:320-327), or in a qualitative manner, such as "pathological complete response" (pCR), "clinical complete response" (cCR), "clinical partial response" (cPR), "clinical stable disease" (cSD), "clinical progressive disease" (cPD), or other qualitative criteria. Tumor response assessments may be performed early after initiation of treatment, for example, after hours, days, weeks, or preferably, after months. Typical endpoints for response assessments are at the end of treatment or at the time of surgical removal of residual tumor cells and / or tumor bed.

[0079] Additional criteria for evaluating response to anticancer treatment relate to survival time, including all of the following: survival time to death, also known as overall survival time (death can be either all-cause or tumor-related); "recurrence-free survival" (the term recurrence includes both local and distant recurrence); metastasis-free survival; disease-free survival (the term disease includes cancer and its associated diseases). Such survival time length can be calculated by reference to a defined starting point (e.g., time of diagnosis or start of treatment) and end point (e.g., death, recurrence, or metastasis). Furthermore, the criteria for the effectiveness of treatment can be expanded to include response to chemotherapy, survival rate, metastasis rate within a given period, and tumor recurrence rate.

[0080] In some embodiments, the CEACAM1-binding antibody is administered in combination with one or more additional anti-cancer / anti-tumor agents or agents. In this context, "anti-tumor agent" or equivalently "cancer agent" refers to an agent administered to a subject for the purpose of treating cancer. Treating cancer can include preventing the onset or progression of cancer, reducing symptoms of cancer, and / or inhibiting the growth of established cancer. Various types of agents for treating cancer are described herein and / or known in the art. For the purposes of this specification, cancer agents can be classified as chemotherapeutic agents, immunotherapeutic agents, cancer vaccines, hormonal therapy agents, and biological response modifiers.

[0081] Cancer drugs function in a variety of ways. Some cancer drugs work by targeting physiological mechanisms specific to tumor cells. Examples include targeting specific genes and their gene products (primary proteins) that are mutated or over- or inappropriately expressed in cancer. Such genes include, but are not limited to, oncogenes (e.g., Ras, Her2, bcl-2), tumor suppressor genes (e.g., EGF, p53, Rb), and cell cycle targets (e.g., CDK4, p21, telomerase). Cancer drugs may alternatively target signaling pathways and molecular mechanisms that are altered in cancer cells. Targeting cancer cells via epitopes expressed on their cell surface is achieved, for example, through the use of monoclonal antibodies.

[0082] Other cancer drugs target cells other than cancer cells. For example, some drugs stimulate the immune system to attack tumor cells (e.g., cancer vaccines and immune checkpoint inhibitors). While this section concerns therapies that may be combined with anti-CEACAM1 antibodies, it should be noted that anti-CEACAM1 therapies promote anti-cancer immune responses by interfering with the immunosuppressive function of CEACM1 and are therefore a type of immunotherapy. Still other drugs, such as angiogenesis inhibitors, work by attacking the blood supply to solid tumors. Angiogenesis inhibitors include basic FGF (b-FGF), VEGF, angiopoietin, angiostatin, endostatin, TNF-α, TNP-470, thrombospondin-1, platelet factor 4, CAI, and certain members of the integrin protein family. Because most malignant cancers are capable of metastasis (i.e., breaking away from the primary tumor site and taking root in distant tissues, thereby forming secondary tumors), drugs that prevent this metastasis are also useful in the treatment of cancer. One category of such drugs is the metalloproteinase inhibitors, which block enzymes used by cancer cells to detach from the primary tumor site and extravasate into other tissues.

[0083] As used herein, chemotherapeutic agents include all other forms of cancer drugs that do not fall into the category of immunotherapeutic agents or cancer vaccines. As used herein, chemotherapeutic agents include both chemical and biological agents. These agents function to inhibit the cellular activity that cancer cells or tumors depend on for continued survival. Categories of chemotherapeutic agents include alkylating / alkaloid agents, antimetabolites, hormones or hormone analogs, and various anti-neoplastic agents. Most, if not all, of these agents are directly toxic to cancer cells and do not require immune stimulation.

[0084] Chemotherapeutic agents currently in development or in clinical use include, among others, 5-FU enhancer, 9-AC, AG2037, AG3340, aggrecanase inhibitors, aminoglutethimide, amsacrine (m-AMSA), angiogenesis inhibitors, anti-VEGF, asparaginase, azacytidine, batimastat (BB94), BAY 12-9566, BCH-4556, bis-naphthalimide, busulfan, capecitabine, carboplatin, Carmustaine+ Poriflosan, cdk4 / cdk2 inhibitors, Chlorombucil, CI-994, Cisplatin, Cladribine, CS-682, Cytarabine HCl, D2163, Dactinomycin, Daunorubicin HCl, DepoCyt, Dexifosamide, Docetaxel, Dolastain, Doxifluridine, Doxorubicin, DX8951f, E 7070, EGFR, Epirubicin, Erythropoietin, Estramustine phosphate sodium, Etoposide (VP16-213), Farnesyltransferase inhibitors, FK 317, Flavopiridol, Floxuridine, Fludarabine, Fluorouracil (5-FU), Flutamide, Flagyline, Gemcitabine, Hexamethylmelamine (HMM), Hydroxyurea (Hydroxycarbamide), Ifosfamide, Interferon alpha-2a, Interferon alpha-2b, Interleukin-2, Irinotecan, ISI 641, Krestin, Lemonal DP 2202, Leuprolide acetate (LHRH releasing factor analog), Levamisole, LiGLA (Lithium gamma linolenic acid), Lodine Seed, Lometexol, Lomustine (CCNU), Marimistat, Mechlorethamine HCl (Nitrogen mustard), Megestrol acetate, Megalamine GLA, mercaptopurine, mesna, mitoguazone (methyl-GAG; methylglyoxal bis-guanylhydrazone; MGBG), mitotane (o.p'-DDD), mitoxantrone, mitoxantrone HCl, MMI 270, MMP, MTA / LY 231514, octreotide, ODN 698, OK-432, oral platinum, oral taxoids, paclitaxel (TAXOL®), PARP inhibitors, PD 183805, pentostatin (2' deoxycoformycin), PKC 412, plicamycin, procarbazine HCl, PSC 833, ralitrexed, RAS Famesyl transferase inhibitors, RAS oncogene inhibitors, semustine (methyl-CCNU), streptozocin, suramin, tamoxifen citrate, taxane analogs, temozolomide, teniposide (VM-26), thioguanine, thiotepa, topotecan, tyrosine kinase, UFT (tegafur / uracil), valrubicin, VEGF / b-FGF inhibitors, vinblastine sulfate, vindesine sulfate, VX-710, VX-853, YM 116, ZD 0101, ZD 0473 / Anormed, ZD 1839, and ZD9331.

[0085] In some preferred embodiments, the subject has an infectious disease or is at increased risk of having an infectious disease. An infectious disease is a disease or condition that results from the presence in a host of a foreign organism or agent that reproduces within the host. An infectious disease typically involves the breaching of normal mucosa or other tissue barriers by an infectious organism or agent. A subject with an infectious disease is a subject with an objectively measurable infectious organism or agent present in the subject's body. A subject at increased risk of having an infectious disease is a subject that is prone to developing an infectious disease. Such subjects may include, for example, subjects with known or suspected exposure to an infectious organism or agent. A subject at increased risk of having an infectious disease may also include subjects with conditions associated with impaired ability to mount an immune response against an infectious organism or agent, such as subjects with congenital or acquired immune deficiencies, subjects undergoing radiation therapy or chemotherapy, subjects with burns, subjects with trauma, subjects undergoing surgery or other invasive medical or dental procedures.

[0086] Infectious diseases are broadly classified as bacterial, viral, fungal, or parasitic based on the category of infectious organism or agent involved. Other less common types of infectious diseases are also known in the art, including, for example, infections with rickettsia, mycoplasma, and agents that cause scrapie, bovine spongiform encephalopathy (BSE), and prion diseases (e.g., kuru and Creutzfeldt-Jakob disease). Examples of bacteria, viruses, fungi, and parasites that cause infectious diseases are well known in the art. Infections can be acute, subacute, chronic, or latent, and they can be localized or systemic. In addition, infections can occur primarily intracellularly or extracellularly during at least one phase of the life cycle of the infectious organism or agent in the host.

[0087] Bacteria include both gram-negative and gram-positive bacteria. Examples of gram-positive bacteria include, but are not limited to, Pasteurella species, Staphylococci species, and Streptococcus species. Examples of gram-negative bacteria include, but are not limited to, Escherichia coli, Pseudomonas species, and Salmonella species. Specific examples of infectious bacteria include Helicobacter pylori, Borrelia burgdorferi, Legionella pneumophilia, Mycobacteria species (e.g., M. tuberculosis, M. avium, M. intracellulare, M. kansasii, Mycobacterium gordonae (M.gordonae), Staphylococcus aureus, Neisseria gonorrhoeae, Neisseria meningitidis, Listeria monocytogenes, Streptococcus pyogenes (group A streptococci), Streptococcus agalactiae (group B streptococci), Streptococcus (green group streptococci), Streptococcus faecalis, Streptococcus bovis, Streptococcus (anaerobic species), Streptococcus pneumoniae, pathogenic Campylobacter species, Enterococcus species, Haemophilus influenzae, Bacillus anthracis, Corynebacterium diphtheriae diphtheriae, Corynebacterium spp., Erysipelothrix rhusiopathiae, Clostridium perfringens, Clostridium tetani, Enterobacter aerogenes, Klebsiella pneumoniae, Pasteurella multocida, Bacteroides spp., Fusobacterium nucleatum, Streptobacillus moniliformis, Treponema pallidum, Treponema parthenae Examples of bacteria that may be present include, but are not limited to, Bacillus subtilis, Bacillus pertenue, Leptospira, Rickettsia, and Actinomyces israelii.

[0088] Examples of viruses that have been shown to cause infectious diseases in humans include Retroviridae (e.g., human immunodeficiency viruses, such as HIV-1 (also called HTLV-III), HIV-2, LAV, or HTLV-III / LAV, or other isolates such as HIV-III, and HIV-LP; Picornaviridae (e.g., poliovirus, hepatitis A virus; enterovirus, human coxsackievirus, rhinovirus, echovirus); Caliciviridae (e.g., strains that cause gastroenteritis); Togaviridae (e.g., equine encephalitis virus, rubella virus); Flaviviridae (e.g., dengue virus, encephalitis virus, yellow fever virus); Coronaviridae (e.g., coronavirus); Rhabdoviridae (e.g., vesicular stomatitis virus, rabies virus); Filoviridae (e.g., Ebola virus); Paramiviridae (e.g., rabies virus, rabies virus); Xoviridae (e.g., parainfluenza virus, mumps virus, measles virus, respiratory syncytial virus); Orthomyxoviridae (e.g., influenza virus); Bunyaviridae (e.g., Hantavirus, Bungavirus, Phlebovirus, and Nairovirus); Arenaviridae (hemorrhagic fever viruses); Reoviridae (e.g., reovirus, orbiviurs, and rotavirus); Birnaviridae; Hepadnaviridae (hepatitis B virus); Parvoviridae (parvovirus); Papovaviridae (papillomavirus, polyomavirus); Adenoviridae (most adenoviruses); Herpesviridae (herpes simplex virus (HSV) 1 and 2, Varicella-Zoster Virus, Cytomegalovirus (CMV), Herpesvirus; Poxviridae (Variolavirus, Vaccinia Virus, Poxvirus); and Iridoviridae (e.g., African Swine Fever Virus); and unclassified viruses (e.g., pathogenic agents of spongiform encephalopathies, agents of delta hepatitis (thought to be a defective satellite of Hepatitis B Virus), non-A, non-B hepatitis agents (Class 1 = enterally transmitted; Class 2 = parenterally transmitted (i.e., Hepatitis C); Norwalk and related viruses and Astrovirus).

[0089] Examples of fungi include Aspergillus species, Blastomyces dermatitidis, Candida albicans, other Candida species, Coccidioides immitis, Cryptococcus neoformans, Histoplasma capsulatum, Chlamydia trachomatis, Nocardia species, and Pneumocystis carinii.

[0090] Parasites include Anaplasma phagocytophilum, Babesia microti, Babesia divergens, Entamoeba histolytica, Giardia lamblia, Leishmania tropica, Leishmania spp., Leishmania braziliensis, Leishmania donovani, Plasmodium falciparum, Plasmodium malariae, Plasmodium ovale, Plasmodium vivax, and Toxoplasma gondii, Trypanosoma gambiense, and other parasites. gambiense and Trypanosoma rhodesiense (African sleeping sickness), Trypanosoma cruzi (Chagas disease), and blood-borne and / or tissue parasites such as Toxoplasma gondii, flatworms, and roundworms.

[0091] In some embodiments, the methods described herein further comprise administering an antibacterial agent to the subject. An antibacterial agent is an agent that stops or inhibits the growth or function of bacteria. Antibacterial agents that are effective in killing or inhibiting a wide range of bacteria are often referred to as broad-spectrum antibiotics. Other types of antibacterial agents are primarily effective against gram-positive or gram-negative bacteria. These types of antibacterial agents are often referred to as narrow-spectrum antibiotics. Other antibacterial agents that are effective against a single organism or disease and not against other types of bacteria are often referred to as limited-spectrum antibiotics. Antibacterial agents may be classified based on their primary mode of action. In general, antibacterial agents are cell wall synthesis inhibitors, cell membrane inhibitors, protein synthesis inhibitors, nucleic acid synthesis or function inhibitors, and / or competitive inhibitors.

[0092] Antibacterial agents useful in the methods described herein include natural penicillins, semi-synthetic penicillins, clavulanic acid, cephalolsporins, bacitracin, ampicillin, carbenicillin, oxacillin, azlocillin, mezlocillin, piperacillin, methicillin, dicloxacillin, nafcillin, cephalothin, cephapirin, cephalexin, cefamandole, cefaclor, cefazolin, 19mmune19al19, cefoxitin, cefotaxime, cefsulodin, cefetamet, cefixime, ceftriaxone, cefoperazone, 19mmune19al19, cephalosporin ... mmune19al19r, moxalactam, carbapenem, imipenem, monobactem, euztreonam, vancomycin, polymyxin, amphotericin B, nystatin, imidazole, clotrimazole, miconazole, ketoconazole, itraconazole, fluconazole, rifampin, ethambutol, tetracycline, chloramphenicol, macrolides, aminoglycosides, streptomycin, kanamycin, tobramycin, amikacin, gentamicin, tetracycline, minocycline, doxycycline, chlortetracycline, erythromycin, roxithromycin, clarithromycin, oleandomycin, azithromycin, chloramphenicol, quinolone, cotrimoxazole, norfloxacin, cytomycin Profloxacin, enoxacin, nalidixic acid, temafloxacin, sulfonamides, gantridine, and trimethoprim; Acedapsone; Acetosulfonate sodium; Alamecin; Alexidine; Amdinocillin; Amdinocillin pivoxil; Amicycline; Amifloxacin; Amifloxacin mesylate; Amikacin; Amikacin sulfate; Aminosalicylic acid; Sodium aminosalicylate; Amoxicillin; Amphomycin; Ampicillin; Ampicillin sodium; Apalcillin sodium; Apramycin; Aspartocin; Astromycin sulfate; Avilamycin; Avoparcin; Azithromycin; Azlocillin; Azlocillin sodium; Bacampicillin hydrochloride; Bacitracin; Bacitracin methylene disalicylate; Bacitracin zinc; Bamberimycin; Benzoylpas calcium;Berethromycin;Betamycin sulfate;Biapenem;Vinillamycin;Biphenamine hydrochloride;Bispyrithione magsulfex;Buticacin;Butirosin sulfate;Capreomycin sulfate;Carbadox;Carbenicillin disodium;Carbenicillin indanyl sodium;Carbenicillin phenylsodium;Carbenicillin potassium;Carumonamu sodium;Cefaclor;Cefadroxil;Cefamandole;Cefamandole naphate;Cefamandole sodium;Cefaparol;Cefatridine;Cefazaflur sodium; Cefazolin;Cefazolin sodium;Cefbuperazone;Cefdinir;Cefepime;Cefepime hydrochloride;Cefetcol;Cefixime;Cefmenoxime hydrochloride;Cefmetazole;Cefmetazole sodium;Cefonicid monosodium;Cefonicid sodium;Cefoperazone sodium;Ceforanide;Cefotaxime sodium;Cefotetan;Cefotetan disodium;Cefotiam hydrochloride;Cefoxitin;Cefoxitin sodium;Cefpimizole;Cefpimizole sodium;Cefpiramide;Cefpiramide sodium;Cefpirime Sulfate;Cefpodoxime proxetil;Cefprozil;Cefroxadine;Cefsulodin sodium;Ceftazidime;Ceftibuten;Ceftizoxime sodium;Ceftriaxone sodium;Cefuroxime;Cefuroxime axetil;Cefuroxime pivoxetil;Cefuroxime sodium;Cephacetrile sodium;Cephalexin;Cephalexin hydrochloride;Cephaloglycine;Cephaloridine;Cephalothin sodium;Cephapirin sodium;Cephradine;Cetocycline hydrochloride;Cetofenicol;Chloramphenicol;Chlorapalmitate Muphenicol;Chloramphenicol pantothenate complex;Chloramphenicol sodium succinate;Chlorhexidine phosphanilate;Chloroxylenol;Chlortetracycline bisulfate;Chlortetracycline hydrochloride;Cinoxacin;Ciprofloxacin;Ciprofloxacin hydrochloride;Ciloremycin;Clarithromycin;Clinafloxacin hydrochloride;Clindamycin;Clindamycin hydrochloride;Clindamycin palmitate hydrochloride;Clindamycin phosphate;Clofazimine;Cloxacillin benzathine;Cloxacillin sodium;Cloxiquin;Colistimethate sodium;Colistin sulfate;Coumarmycin;Coumarmycin sodium;Ciclacillin;Cycloserine;Dalfopristin;Dapsone;Daptomycin;Demeclocycline;Demeclocycline hydrochloride;Demecycline;Denofungin;Diaveridine;Dicloxacillin;Dicloxacillin sodium;Dihydrostreptomycin sulfate;Dipyrithione;Dirithromycin;Doxycycline;Doxycycline calcium;Doxycycline phosphatex;Doxycycline hyclate;Droxa Synthin sodium;Enoxacin;Epicillin;Epitetracycline hydrochloride;Erythromycin;Erythromycin acistrate;Erythromycin estolate;Erythromycin ethylsuccinate;Erythromycin gluceptate;Erythromycin lactobionate;Erythromycin propionate;Erythromycin stearate;Ethambutol hydrochloride;Ethionamide;Fleroxacin;Floxacillin;Fludalanine;Flumequine;Fosfomycin;Fosfomycin tromethamine;Fumoxicillin;Furazolium chloride;Furazolium tartrate Zolium;Sodium fusidate;Fusidic acid;Gentamicin sulfate;Glucoximide;Gramicidin;Haloprogin;Hetacillin;Hetacillin potassium;Hexedin;Ibafloxacin;Imipenem;Isoconazole;Isepamicin;Isoniazid;Josamycin;Kanamycin sulfate;Kitasamycin;Levofuraltadone;Levopropylcillin potassium;Lexitromycin;Lincomycin;Lincomycin hydrochloride;Lomefloxacin;Lomefloxacin hydrochloride;Lomefloxacin mesylate;Loracarbef;Mafenide;Meclocycline ;Meclocycline sulfosalicylate;Megalomycin potassium phosphate;Mequidox;Meropenem;Methacycline;Methacycline hydrochloride;Methenamine;Methenamine hippurate;Methenamine mandelate;Methicillin sodium;Methioprim;Metronidazole hydrochloride;Metronidazole phosphate;Mezlocillin;Mezlocillin sodium;Minocycline;Minocycline hydrochloride;Milinkamycin hydrochloride;Monensin;Monensin sodium;Nafcillin sodium;Nalidixic acid sodium;Nalidixic acid;Natamycin;Nebramycin;Neomycin palmitate;Neomycin sulfate;Neomycin undecylenate;Netilmicin sulfate;Neuthramycin;Nifuraden;Nifuraldeson;Nifuratel;Nifuratron;Nifurdazil;Nifrimide;Nifurpirinol;Nifluquinazole;Nifurthiazole;Nitrocycline;Nitrofurantoin;Niromide;Norfloxacin;Novobiocin sodium;Ofloxacin;Ormetoprim;Oxacillin sodium;Oxomonam;Oxomonam sodium;Oxolinic acid;Oxytetracycline;Oxytetracycline Calcium;Oxytetracycline hydrochloride;Paldimycin;Parachlorophenol;Pauromycin;Pefloxacin;Pefloxacin mesylate;Penamecillin;Penicillin G benzathine;Penicillin G potassium;Penicillin G procaine;Penicillin G sodium;Penicillin V;Penicillin V benzathine;Penicillin V hydrabamine;Penicillin V potassium;Pentidone sodium;Phenyl aminosalicylate;Piperacillin sodium;Pirbenicillin sodium;Pyridicillin sodium;Pirlimycin hydrochloride;Pivampicillin hydrochloride;Pi Vampicillin pamoate;Pivampicillin probenate;Polymyxin B sulfate;Porfiromycin;Propicacin;Pyrazinamide;Zinc pyrithione;Quindecamin acetate;Quinupristin;Racephenicol;Ramoplanin;Ranimycin;Relomycin;Repromycin;Rifabutin;Rifamethane;Rifamexyl;Rifamide;Rifampin;Rifapentine;Rifaximin;Rolitetracycline;Rolitetracycline nitrate;Rosaramycin;Rosaramycin butyrate;Rosaramycin propionate;Rosaramycin sodium phosphate ;Rosaramicin stearate;Rosoxacin;Roxarsone;Roxithromycin;Sancycline;Sanfetrinemucine sodium;Salmoxicillin;Salpicillin;Scopafungin;Sisomicin;Sisomicin sulfate;Sparfloxacin;Spectinomycin hydrochloride;Spiramicin;Stalimycin hydrochloride;Stephimycin;Streptomycin sulfate;Streptonicozide;Sulfabenz;Sulfabenzamide;Sulfacetamide;Sulfacetamide sodium;Sulfacytine;Sulfadiazine;Sulfadiazine sodium;Sulfadoxine;Sulfalene;Sulfamerazine;Sulfameta;Sulfamethazine;Sulfamethizole;Sulfamethoxazole;Sulfamonomethoxine;Sulfamoxole;Zinc sulfanilate;Sulfanitran;Sulfasalazine;Sulfasomizole;Sulfasomizole;Sulfazameth;Sulfisoxazole;Acetylsulfisoxazole;Sulfisoxazolediolamine;Sulfomixin;Sulopenem;Sultamicillin;Santamicin sodium;Talampicillin hydrochloride;Teicoplanin;Temafloxacin hydrochloride;Temocillin;Tetracycline;Te These include, but are not limited to, tracycline hydrochloride; tetracycline metaphosphate; tetroxoprim; thiamphenicol; tiphencillin potassium; ticarcillin clecil sodium; ticarcillin disodium; ticarcillin monosodium; ticlatone; thiodonium chloride; tobramycin; tobramycin sulfate; tosufloxacin; trimethoprim; trimethoprim sulfate; trisulfapyrimidine; troleandomycin; trospectomycin sulfate; tyrothricin; vancomycin; vancomycin hydrochloride; pristinamycin; and zorbamycin.;

[0093] In some embodiments, the method further comprises administering an antiviral agent to the subject.Antiviral agents are compounds that prevent the infection of cells by viruses or the replication of viruses in cells.Compared to antibacterial agents, there are far fewer antiviral agents, because the process of viral replication is closely related to DNA replication in host cells, and non-specific antiviral agents are often assumed to be toxic to hosts.In the process of viral infection, there are several steps that can be blocked or inhibited by antiviral agents.These steps include virus attachment to host cells (immunoglobulins or binding peptides), virus uncoating (e.g., amantadine), viral mRNA synthesis or translation (e.g., interferon), viral RNA or DNA replication (e.g., nucleoside analogs), new viral protein maturation (e.g., protease inhibitors), and virus budding and release.

[0094] Nucleotide analogues are synthetic compounds that are similar to nucleotides but have incomplete or abnormal deoxyribose or ribose groups. When nucleotide analogues enter cells, they are phosphorylated to form triphosphates that compete with normal nucleotides for incorporation into viral DNA or RNA. When the triphosphate form of nucleotide analogues is incorporated into a growing nucleic acid chain, it causes irreversible binding with viral polymerase, thus terminating the chain. Nucleotide analogues include, but are not limited to, acyclovir (used to treat herpes simplex virus and varicella zoster virus), ganciclovir (useful for treating cytomegalovirus), idoxuridine, ribavirin (useful for treating respiratory syncytial virus infections), dideoxyinosine, dideoxycytidine, and zidovudine (azidothymidine).

[0095] Interferons are cytokines secreted by virus-infected cells and immune cells. They function by binding to specific receptors on infected and neighboring cells, causing the cells to undergo changes that protect them from infection by the virus. IFN-α and IFN-β also induce the expression of class I and class II MHC molecules on the surface of infected cells, increasing antigen presentation for host immune cell recognition. These interferons are available in recombinant form and are used to treat chronic hepatitis B and hepatitis C infections. At doses effective for antiviral therapy, interferons can have severe side effects such as fever, fatigue, and weight loss.

[0096] Immunoglobulin therapy is used to prevent viral infection. Immunoglobulin therapy for viral infection is different from bacterial infection because it is not antigen specific and it works by binding to extracellular virions and preventing them from attaching and entering cells susceptible to viral infection. The therapy is effective in preventing viral infection for the period of time that the antibodies are present in the host. In general, there are two types of immunoglobulin therapy: normal immunoglobulin therapy and hyperimmune globulin therapy. Normal immunoglobulin therapy utilizes pooled antibody products prepared from the serum of normal blood donors. The pooled products contain low titers of antibodies against a wide range of human viruses, such as Hepatitis A, Parvovirus, and Enterovirus (especially in newborns). Hyperimmune globulin therapy utilizes antibodies prepared from the serum of individuals who have high titers of antibodies against a particular virus. These antibodies are then used against a particular virus. Examples of hyperimmune globulins include varicella zoster immune globulin (useful in preventing chickenpox in immunocompromised children and newborns), human rabies immune globulin (useful in post-exposure prophylaxis of subjects bitten by a rabid animal), hepatitis B immune globulin (particularly useful in preventing hepatitis B virus in subjects exposed to the virus), and RSV immune globulin (useful in treating respiratory syncytial virus infections).

[0097] Thus, antiviral agents useful in combination with the anti-CEACAM1 antibody agents described herein include, but are not limited to, immunoglobulins, amantadine, interferons, nucleoside analogs, and protease inhibitors. Specific examples of antiviral agents include acemannan; acyclovir; acyclovir sodium; adefovir; alovudine; alvirceptosudotox; amantadine hydrochloride; aranotin; aryldone; atevirdine mesylate; avridine; cidofovir; cipamfilline; cytarabine hydrochloride; delavirdine mesylate; desciclovir; didanosine; disoxalyl; edoxudine; enviraden; enviroxime; famciclovir; famotine hydrochloride; fiacitabine; fiaruridine; fosarilate; foscarnet sodium; phosphonet sodium; ganciclovir; ceciclovir; cecitabine; cecitabine; ceciclovir ... These include, but are not limited to, clovir; ganciclovir sodium; idoxuridine; kethoxal; lamivudine; lobucavir; memotin hydrochloride; methisazone; molnupiravir; nevirapine; nilmatrellvir; penciclovir; pirodavir; ribavirin; rimantadine hydrochloride; ritonavir; saquinavir mesylate; somantadine hydrochloride; sorivudine; statolone; stavudine; tilorone hydrochloride; trifluridine; valacyclovir hydrochloride; vidarabine; vidarabine phosphate; vidarabine sodium phosphate; viloxime; zalcitabine; zidovudine; and vinviroxime.

[0098] In some embodiments, the methods described herein further comprise administering an antifungal agent to the subject. An antifungal agent is an agent that stops or inhibits the growth or function of infectious fungi. Antifungal agents may be classified by their mechanism of action. Some antifungal agents function as cell wall inhibitors by inhibiting glucose synthase. These include, but are not limited to, basiungin / ECB. Other antifungal agents function by destabilizing membrane integrity. These include, but are not limited to, imidazoles such as clotrimazole, sertaconzole, fluconazole, itraconazole, ketoconazole, miconazole, and voriconacole, as well as FK 463, amphotericin B, BAY 38-9502, MK 991, pradimicin, UK 292, butenafine, and terbinafine. Other antifungal agents work by breaking down chitin (e.g., chitinase) or by immunosuppression (501 cream).

[0099] Accordingly, antifungal agents for use in the methods described herein include imidazole, 501 cream, and acrisorcin, ambruticin, amorolf, amphotericin B, azaconazole, azaserine, vadifungin, BAY 38-9502, bifonazole, biphenamine hydrochloride, bispyrithione magsulfex, butenafine, butoconazole nitrate, calcium undecylenate, candicidin, carbol fuchsin, chitinase, chlordantoin, ciclopirox, ciclopirox olamine, cilofungin, cisconazole, clotrimazole, cuprimixin, denofungin, dipyrithione, doconazole, econazole, econazole nitrate, enilconazole, ethnam nitrate, fenticonazole nitrate, filipin, FK 463, fluconazole, flucytosine, fungimycin, griseofulvin, hamycin, isoconazole, itraconazole, kalafungin, ketoconazole, romofungin, lyzimycin, mepartricin, miconazole, miconazole nitrate, MK 991, monensin, monensin sodium, naftifine hydrochloride, neomycin undecylenate, nifuratel, niflumerone, nitralamine hydrochloride, nystatin, octanoic acid, orconazole nitrate, oxiconazole nitrate, oxyfungin hydrochloride, parconazole hydrochloride, partricin, potassium iodide, pradimicin, proclonol, zinc pyrithione, pyrrolnitrin, rutamycin, sanguinarium chloride, saperconazole, scopafungin, selenium sulfide, sertaconazole, sinefungin, sulconazole nitrate, terbinafine, terconazole, thiram, ticlatone, tioconazole, tolciclate, tolindate, tolnaftate, triacetin, triafungin, UK 292, undecylenic acid, viridofulvin, voriconazole, zinc undecylenate, and dinoconazole hydrochloride.

[0100] In some embodiments, the methods described herein further comprise administering to the subject an anti-parasitic drug.Anti-parasitic drugs are agents that stop or inhibit the growth or function of infectious parasites.Anti-parasitic drugs are also referred to as parasiticides, and examples of anti-parasitic drugs useful in the methods described herein include albendazole, amphotericin B, benznidazole, bithionol, chloroquine HCl, chloroquine phosphate, clindamycin, dehydroemetine, diethylcarbamazine, diloxanide furoate, doxycycline, eflornithine, furazolidaone, glucocorticoids, halofantrine, iodoquinol, ivermectin, mebendazole, mefloquine, meglumine antimoniate, melarsoprol, metrifonate, metronidazole, niclosamide, nifurtimox, oxamniquine, These include, but are not limited to, paromomycin, pentamidine isethionate, piperazine, praziquantel, primaquine phosphate, proguanil, pyrantel pamoate, pyrimethammine sulfonamide, pyrimethammine sulfadoxine, quinacrine HCl, quinine sulfate, quinidine gluconate, spiramycin, sodium stibogluconate (antimony sodium gluconate), suramin, tetracycline, thiabendazole, tinidazole, trimesroprim-sulfamethoxazole, and triparthamide, some of which may be used alone or in combination with others.

[0101] Conditions that may benefit from anti-hCEACAM1 monoclonal antibody therapy include those that require enhanced B and T cell responses to immunogens, such as melanoma, pancreatic cancer, thyroid cancer, lung cancer, colorectal cancer, squamous cell carcinoma, prostate cancer, breast cancer, bladder cancer, and gastric cancer.

[0102] antibody The antibody that specifically binds to CEACAM1 can be used to inhibit CEWACAM1 activity, thereby treating diseases or disorders in which CEACAM1 is involved in the pathology. The term "antibody" is used in a broad sense and includes monoclonal antibodies (including full-length or intact monoclonal antibodies), polyclonal antibodies, multivalent antibodies, multispecific antibodies (e.g., bispecific antibodies), antibody fragments, and antigen-binding portions thereof (e.g., paratopes, CDRs), so long as they exhibit the desired biological activity and specificity.

[0103] As used herein, "antibody variable domain" refers to the portion of the light and heavy chains of an antibody molecule that contains the amino acid sequences of the complementarity determining regions (CDRs; i.e., CDR1, CDR2, and CDR3) and framework regions (FRs). VH refers to the variable domain of the heavy chain. VL refers to the variable domain of the light chain. The amino acid positions assigned to the CDRs and FRs may be defined according to the numbering conventions of Kabat or Chothia. The term "framework region" (FR) refers to the variable domain residues other than the CDR residues. (Kabat et al., Sequences of Proteins of Immunological Interest, 5 th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1987, 1991)) or Chothia (Chothia & Lesk 196 J. Mol. Biol. 901 (1987)).

[0104] The "complementarity determining regions" (CDRs) of an antibody are those portions of an antibody variable domain that are (typically) involved in antigen binding. Each variable domain typically has three CDR regions identified as CDR1, CDR2, and CDR3. Each CDR may, for example, include amino acid residues from the CDRs defined by Kabat (i.e., approximately residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) in the light chain variable domain and 31-35 (H1), 50-65 (H2), and 95-102 (H3) in the heavy chain variable domain). Each CDR may also contain amino acid residues from the "hypervariable loops" (i.e., approximately residues 26-32 (LI), 50-52 (L2), and 91-96 (L3) in the light chain variable domain and 26-32 (H1), 53-55 (H2), and 96-101 (H3) (Chothia) in the heavy chain variable domain). In some cases, a CDR may contain amino acids from both the CDR regions and the hypervariable loops defined according to Kabat. The Kabat residue designations do not always correspond directly with the linear numbering of the amino acid residues (primary amino acid sequence). The actual linear amino acid sequence, whether framework or CDR, may contain fewer or additional amino acids compared to those in the strict Kabat numbering due to shortening of or insertion into structural elements of the basic variable domain structure. The correct Kabat residue numbering can be determined for a given antibody or antigen-binding fragment thereof by aligning the homologous residues in the sequence of the antibody or antigen-binding fragment thereof with the "standard" Kabat numbering sequence. An example of how the Kabat numbering relates to the primary amino acid sequence of an antibody can be found in PCT / US2019 / 065212, which is incorporated by reference in its entirety. Alternatively, the CDRs can be defined according to the ImMunoGeneTics (IMGT) system (Lefranc, M.-P. et al., Dev. Comp. Immunol., 27, 55-77 (2003)).

[0105] In one embodiment, an antibody or antigen-binding fragment thereof that binds to the GF-CC' face of a CEACAM1 protein comprises a heavy chain variable region and a light chain variable region, each of which comprises a CDR1, a CDR2, and a CDR3, and the sequence of CDR1 (CDR1H) of the heavy chain variable region comprises the sequence X1HX2X3S (SEQ ID NO:1); X1 is A, D, N, or S; X2 is A or G; and X3 is an amino acid having a hydrophobic side chain, including I or M; and the sequence of CDR2 (CDR2H) of the heavy chain variable region comprises the sequence TIFF2025506405000015.tif4128; the sequence of the CDR3 (CDR3H) of the heavy chain variable region is X4 is D, G, or P; X5 is F or P; X6 is D or F; X7 is A or Y; and X8 is L, H, or F; the sequence of the CDR1 (CDR1L) of the light chain variable region comprises the sequence RANSAVSYMY (SEQ ID NO:4); the sequence of the CDR2 (CDR2L) of the light chain variable region comprises the sequence LTSNRAT (SEQ ID NO:5); and the sequence of the CDR3 (CDR3L) of the light chain variable region comprises the sequence TIFF2025506405000017.tif4128; X9 is W or N; X 10 is S or T;X 11 is A or an amino acid having a neutral hydrophilic side chain, including S, N, and T; X 12 is L, F, or N;X 13 is P or F.

[0106] In one embodiment, the region comprises SEQ ID NO:2, the sequence of CDR3 of the heavy chain variable region comprises SEQ ID NO:8; the sequence of CDR1 of the light chain variable region comprises SEQ ID NO:4; the sequence of CDR2 of the light chain variable region comprises SEQ ID NO:5; and the sequence of CDR3 of the light chain variable region comprises SEQ ID NO:10.

[0107] In one embodiment, the CEACAM1 antibody or antigen-binding fragment thereof comprises six CDRs, wherein the sequence of CDR1 of the heavy chain variable region comprises SEQ ID NO:7; the sequence of CDR2 of the heavy chain variable region comprises SEQ ID NO:2; the sequence of CDR3 of the heavy chain variable region comprises SEQ ID NO:8; the sequence of CDR1 of the light chain variable region comprises SEQ ID NO:14; the sequence of CDR2 of the light chain variable region comprises SEQ ID NO:5; and the sequence of CDR3 of the light chain variable region comprises SEQ ID NO:9.

[0108] According to certain embodiments, the antibodies and antigen-binding fragments thereof also feature humanized frameworks to reduce immunogenicity. In certain embodiments, the CDRs of the antibodies or antigen-binding fragments thereof are located in frameworks derived from human antibodies or antigen-binding fragments thereof. In other embodiments, surface-exposed framework residues of the contemplated antibodies or antigen-binding fragments thereof are replaced with framework residues of human antibodies or antigen-binding fragments thereof. The CDRs may also be located in mouse or humanized frameworks linked to human constant regions (i.e., chimeric antibodies). In preferred embodiments, the CDRs of the antibodies or antigen-binding fragments thereof are located in frameworks that are composites of two or more human antibodies. In such embodiments, the antibodies or antigen-binding fragments thereof contain two or more sequence segments ("composite") from V regions of unrelated human antibodies that have all been filtered, for example using "in silico tools" (Holgate & Baker, IDrugs. 2009 Apr;12(4):233-7), for the presence of potential T-cell epitopes. The precise matching of human sequence segments in all sections of the starting antibody V region and the elimination of CD4+ T cell epitopes prior to the synthesis of the antibody or its antigen-binding fragment allows this technique to limit or avoid immunogenicity while maintaining optimal affinity and specificity through up-front analysis of sequences required for antigen specificity (Holgate & Baker, 2009).

[0109] As used herein, the term "identity" refers to the sequence identity between two nucleic acid molecules or polypeptides. Identity can be determined by comparing the positions in each sequence that can be aligned for comparison purposes. For example, if a position in the nucleotide sequence being compared is occupied by the same base, the molecules are identical at that position. The degree of identity between nucleic acid or amino acid sequences is a function of the number of identical or matching nucleotides or amino acids at shared positions. For example, polypeptides and polynucleotides encoding such polypeptides that have at least 85%, 90%, 95%, 98% or 99% identity with the specific polypeptides described herein and preferably exhibit substantially the same function are contemplated. Methods and computer programs for determining both sequence identity and similarity are publicly available, including, but not limited to, the GCG program package (Devereux et al., Nucleic Acids Research 12: 387, 1984), BLASTP, BLASTN, FASTA (Altschul et al., J. Mol. Biol. 215:403 (1990), and Alignment Programs (version 2.0). The well-known Smith Waterman algorithm may also be used to determine similarity. BLAST programs are publicly available from NCBI and other sources (BLAST Manual, Altschul, et al., NCBI NLM NIH, Bethesda, Md. 20894; BLAST 2.0 at ncbi.nlm.nih.gov / blast / ). In comparing sequences, such methods account for various substitutions, deletions, and other modifications.

[0110] In another aspect, the CEACAM1-binding antibody or antigen-binding fragment thereof that binds to the GF-CC' face of CEACAM1 is (i) a heavy chain variable domain comprising a sequence that is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the heavy chain variable domain sequence of SEQ ID NO:11; and / or (ii) a light chain variable domain comprising a sequence at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the light chain variable domain sequence of SEQ ID NO:12. Includes.

[0111] In some embodiments, amino acid sequence modification(s) of the antibody or antigen-binding fragment thereof described herein that binds to CEACAM1 are contemplated. The amino acid sequence variant of the antibody or antigen-binding fragment thereof is prepared by introducing appropriate nucleotide changes into the nucleic acid encoding the antibody or antigen-binding fragment thereof, or by peptide synthesis. Such modifications include, for example, deletion from and / or insertion into and / or substitution of residues in the amino acid sequence of the antibody or antigen-binding fragment thereof. Any combination of deletion, insertion, and substitution is performed to arrive at the final construct, provided that the final construct has the desired characteristics, for example, binding specificity, inhibition of biological activity, etc.

[0112] One type of variant is a conservative amino acid substitution variant. In such variants, at least one amino acid residue in an antibody or antigen-binding fragment thereof is replaced by a different residue having similar side chain properties. Amino acids can be grouped according to the similarity of their chain properties (see Lehninger, BIOCHEMISTRY (2nd ed., Worth Publishers, New York, 1975)): (1) Non-polar: Ala (A), Val (V), Leu (L), Ile (I), Pro (P), Phe (F), Trp (W), Met (M); (2) no polarity change: Gly (G), Ser (S), Thr (T), Cys (C), Tyr (Y), Asn (N), Gln (Q); (3) Acidic: Asp (D), Glu (E); (4) Basic: Lys (K), Arg (R), His (H). Thus, a non-limiting example of a conservative amino acid substitution is the replacement of a non-polar amino acid with another non-polar amino acid.

[0113] Alternatively, naturally occurring residues can be grouped based on common side chain properties: (1) Hydrophobicity: Ala (A), Val (V), Leu (L), Ile (I), Met (M); (2) Neutral hydrophilicity: Ser (S), Thr (T), Cys (C), Asn (N), Gln (Q); (3) Acidic: Asp (D), Glu (E); (4) Basic: Lys (K), Arg (R), His (H); (5) Residues that affect chain orientation: Gly (G), Pro (P); (6) Aromatic: Phe (F), Trp (W), Tyr (Y). Thus, a non-limiting example of a conservative amino acid substitution is one that replaces a hydrophobic amino acid with another hydrophobic amino acid.

[0114] Amino acid sequence insertions, which may include amino and / or carboxyl terminal fusions, ranging in length from one residue to polypeptides containing 100 or more residues, as well as intrasequence insertions of single or multiple amino acid residues are further contemplated. Examples of terminal insertions include an antibody or antigen-binding fragment thereof with an N-terminal methionyl residue, or an antibody or antigen-binding fragment thereof fused to a cytotoxic polypeptide. Other insertion variants of antibodies or antigen-binding fragments thereof include fusions to the N- or C-terminus of the antibody or antigen-binding fragment thereof with enzymes or polypeptides that increase the serum half-life of the antibody or antigen-binding fragment thereof, such as, for example, biotin.

[0115] Any cysteine ​​residue not involved in maintaining the proper conformation of an antibody or antigen-binding fragment thereof that binds to CEACAM1 can also be substituted, for example with serine or alanine, to improve the oxidative stability of the molecule and prevent aberrant crosslinking.

[0116] Conversely, the addition of cysteine ​​bond(s) to an antibody or antigen-binding fragment thereof can improve its stability (particularly where the antibody or antigen-binding fragment thereof is an antibody fragment such as an Fv fragment).

[0117] In some embodiments, the antibody or antigen-binding fragment thereof has an amino acid alteration that alters the original glycosylation pattern of the antibody or antigen-binding fragment thereof. By "altering the original glycosylation pattern" is intended the deletion of one or more carbohydrate moieties found in the antibody or antigen-binding fragment thereof and / or the addition of one or more glycosylation sites not present in the antibody or antigen-binding fragment thereof. Glycosylation of antibodies is typically either N-linked or O-linked. N-linked refers to the addition of a carbohydrate moiety to the side chain of an asparagine residue. The tripeptide sequences asparagine-X-serine and asparagine-X-threonine are recognition sequences for the enzymatic addition of a carbohydrate moiety to the asparagine side chain, where X is any amino acid except proline. Thus, the presence of either of these tripeptide sequences in a polypeptide creates a potential glycosylation site. O-linked glycosylation refers to the addition of one of the sugars N-aceylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine can also be used. The addition of a glycosylation site to an antibody or antigen-binding fragment thereof that binds to CEACAMI is accomplished by altering the amino acid sequence so that it contains one or more of the above tripeptide sequences (for N-linked glycosylation sites). The alteration can also be made by adding one or more serine or threonine residues (for O-linked glycosylation sites) to the sequence of the original antibody or antigen-binding fragment thereof, or by substituting one or more serine or threonine residues (for O-linked glycosylation sites) into the sequence of the original antibody or antigen-binding fragment thereof.

[0118] In some embodiments, the CEACAM1 antibody or antigen-binding fragment thereof provided herein is deglycosylated or aglycosylated. In some embodiments, the CEACAM1 antibody or antigen-binding fragment thereof lacks a glycosylation site in CDR1 of the variable light chain. In some embodiments, the CEACAM1 antibody or antigen-binding fragment thereof lacks a NXS / T consensus sequence in CDR1 of the variable light chain. In some embodiments, the CEACAM1 antibody or antigen-binding fragment thereof has a mutation at CDR residue 26 and / or 29 (Kabat numbering) of CDR1 of the variable light chain. If the antibody or antigen-binding fragment thereof comprises an Fc region, the carbohydrate(s) added thereto can be altered. For example, antibodies with mature carbohydrate structures that lack fucose added to the Fc region of the antibody or antigen-binding fragment thereof have been described. See, for example, U.S. Patent Publication Nos. 2003 / 0157108; 2004 / 0093621. WO 03 / 011878; U.S. Patent No. 6,602,684 refer to antibodies with bisecting N-acetylglucosamine (G1cNAc) in the carbohydrate attached to the Fc region of the antibody or antigen-binding fragment thereof. WO 97 / 30087 reports antibodies with at least one galactose residue in the oligosaccharide attached to the Fc region of the antibody or antigen-binding fragment thereof. See also WO 98 / 58964; WO 99 / 22764 for antibodies with altered carbohydrate attached to their Fc region.

[0119] In some embodiments, it may be desirable to modify the antibody or antigen-binding fragment thereof that binds to CEACAM1 described herein with respect to effector function, for example, to enhance the antibody-dependent cellular cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC) of the antibody or antigen-binding fragment thereof. This can be achieved by introducing one or more amino acid substitutions into the Fc region of the antibody or antigen-binding fragment thereof. Alternatively or additionally, one or more cysteine ​​residues can be introduced into the Fc region, thereby allowing interchain disulfide bonds in this region. The homodimeric antibody or antigen-binding fragment thereof thus generated may have improved internalization capability and / or increased complement-mediated cell killing and antibody-dependent cellular cytotoxicity (ADCC). See Caron et al., 176 J. Exp. Med. 1191 (1992); Shopes, 148 J. Immunol. 2918 (1992). Homodimeric antibodies with enhanced anti-tumor activity can also be prepared using heterobifunctional cross-linkers as described in Wolff et al., 53 Cancer Res. 2560 (1993). Alternatively, antibodies or antigen-binding fragments thereof can be engineered that have dual Fc regions and may thereby have enhanced complement lysis and ADCC capabilities. See Stevenson et al., 3 Anti-Cancer Drug Design 219 (1989).

[0120] For example, WO 00 / 42072 describes an antibody with improved ADCC function in the presence of human effector cells, which comprises amino acid substitutions in its Fc region.Preferably, the antibody or antigen-binding fragment thereof with improved ADCC comprises substitutions at positions 298, 333, and / or 334 (Eu residue numbering) in the Fc region.Typically, the Fc region that is altered is a human IgG1 Fc region that comprises or consists of substitutions at one, two, or three of these positions.Such substitutions are optionally combined with substitution(s) that increase Clq binding and / or CDC.Substitutions include the Asn297Ala mutation in IgG1 Fc.

[0121] Antibodies with altered Clq binding and / or complement dependent cytotoxicity (CDC) are described in WO 99 / 51642, U.S. Patent Nos. 6,194,551, 6,242,195, 6,528,624, and 6,538,124. The antibodies contain amino acid substitutions at one or more of amino acid positions 270, 322, 326, 327, 329, 313, 333, and / or 334 (Eu residue numbering) of their Fc region.

[0122] WO 00 / 42072 and US Patent Publication No. 2005 / 0014934 describe antibodies with improved binding to fetal Fc receptor (FcRn) and extended half-life. Such antibodies include an Fc region with one or more substitutions therein that improve the binding of the Fc region to CEACAM1. For example, the Fc region can have substitutions at one or more of positions 238, 250, 256, 265, 272, 286, 303, 305, 307, 311, 312, 314, 317, 340, 356, 360, 362, 376, 378, 380, 382, ​​413, 424, 428, or 434 (Eu residue numbering). A preferred Fc region-containing antibody variant with improved CEACAM1 binding comprises one, two or three amino acid substitutions at positions 307, 380 and 434 (Eu residue numbering) of its Fc region. In one embodiment, the antibody or antigen-binding fragment thereof has a 307 / 434 mutation. Engineered antibodies that bind to CEACAM1 with three or more (e.g., four) functional antigen-binding sites are also contemplated. See, for example, U.S. Patent Publication No. US 2002 / 0004587.

[0123] Antibody Fragments and Types In some embodiments of the aspects described herein, the CEACAM1 antibody fragment is a Fab fragment, which comprises or essentially consists of the variable (VL) and constant (CL) domains of the light chain and the variable domain (VH) and first constant domain (CH1) of the heavy chain.

[0124] In some embodiments of the aspects described herein, the CEACAM1 antibody fragment is a Fab' fragment, which refers to a Fab fragment having one or more cysteine ​​residues at the C-terminus of the CH1 domain.

[0125] In some embodiments of the aspects described herein, the CEACAM1 antibody fragment is an Fd fragment comprising or consisting essentially of the VH and CH1 domains.

[0126] In some embodiments of the aspects described herein, the CEACAM1 antibody portion is an Fd' fragment that includes the VH and CH1 domains and one or more cysteine ​​residues at the C-terminus of the CH1 domain.

[0127] Single chain Fv antibody fragments, i.e., scFv antibody fragments, comprise or essentially consist of the VH and VL domains of an antibody, such that these domains are present in a single polypeptide chain.Generally, Fv polypeptides further comprise a polypeptide linker between the VH and VL domains, which allows scFv to form the desired structure for antigen binding.See, for example, Pluckthun, 113 Pharmacology Monoclonal Antibodies 269 (Rosenburg & Moore, eds., Springer-Verlag, New York, 1994).Thus, in some embodiments of the aspects described herein, CEACAM1 antibody fragments are Fv fragments that comprise or essentially consist of the VL and VH domains of a single arm of an antibody.

[0128] In some embodiments of the aspects described herein, the CEACAM1 antibody portion is a diabody comprising two antigen binding sites, including a heavy chain variable domain (VH) linked to a light chain variable domain (VL) in the same polypeptide chain.

[0129] In some embodiments of the aspects described herein, the CEACAM1 antibody portion is a dAb fragment that includes or consists essentially of the VH domain.

[0130] In some embodiments of the aspects described herein, the CEACAM1 antibody portion is an F(ab')2 fragment, which comprises a bivalent fragment comprising two Fab' fragments linked by disulfide bridges in the hinge region.

[0131] Linear antibodies are described in Zapata et al., Protein Engin., 8(10):1057-1062 (1995). Briefly, such antibodies comprise a pair of tandem Fd segments (VH-CH1-VH-CH1), which together with complementary light chain polypeptides form a pair of antigen-binding regions. Linear antibodies can be bispecific or monospecific. In some embodiments of the aspects described herein, the CEACAM1 antibody fragment is a linear antibody comprising a pair of tandem Fd segments (VH-CH1-VH-CH1), which together with complementary light chain polypeptides form a pair of antigen-binding regions.

[0132] Various techniques have been developed and are available for the production of antibody fragments. Traditionally, such fragments were obtained via proteolytic digestion of intact antibodies. See, for example, Morimoto et al., 24 J. Biochem. Biophys. Meths. 107 (1992); Brennan et al., 229 Science 81 (1985). However, such fragments can now be produced directly by recombinant host cells. For example, antibody fragments can be isolated from antibody phage libraries as discussed herein. Alternatively, Fab'-SH fragments can be directly recovered from E. coli and chemically coupled to form F(ab')2 fragments (Carter et al., 1992). According to another technique, F(ab')2 fragments can be directly isolated from recombinant host cell culture. Other techniques for the production of antibody fragments will be apparent to those skilled in the art. In another embodiment, the antibody fragment of choice is a single chain Fv fragment (scFv). See, for example, WO 93 / 16185.

[0133] In one embodiment, the antibody is a bispecific antibody that comprises a complementary region that binds to CEACAM1 and a complementary region that binds to PD-1.

[0134] In one embodiment, the antibody is a bispecific antibody comprising a complementary region that binds to CEACAM1 and a complementary region that binds to PD-Ll.

[0135] Contemplated antibodies or antigen-binding fragments may have any isotype, including all types of constant regions, including IgM, IgG, IgD, and IgE, as well as IgG1, IgG2, IgG3, and IgG4. In one embodiment, human isotype IgG1 is used. In another embodiment, human isotype IgG4 is used. The light chain constant region may be a or K. Antibodies or antigen-binding fragments thereof may contain sequences from multiple classes or isotypes.

[0136] antibody binding The method described herein allows for determining whether a given subject's CEACAM1 (or the subject's cancer CEACAM1) is inhibited by an antibody that binds to the GF-CC' face of CEACAM1. If an antibody binds efficiently to wild type but either does not bind or binds with reduced affinity to a certain allelic variant, it can be predicted based on the CEACAM1 genotype whether a given subject's CEACAM1-mediated disease or disorder is likely to respond (or not) to the antibody, or it is assumed that the dosage or frequency of administration of the antibody should potentially be increased. The human CEACAM1 gene produces 11 isoforms by alternative splicing. Each isoform has one variable (V)-like Ig domain at the amino (N) terminus of the protein. Various isoforms also have two or three constant C2-like Ig domains, with the exception of the CEACAM1-1L and CEACAM1-1S isoforms. Eight CEACAM1 isoforms are tethered to the cell membrane via a transmembrane domain, and three CEACAM1 isoforms (CEACAM1-4C1, CEACAM1-3, and CEACAM1-3C2) lack the transmembrane domain and are secreted. Two isoforms (CEACAM1-3AL and CEACAM1-3AS) have an Alu family repeat (A) between the constant C2-like Ig domain and the transmembrane domain. Transmembrane CEACAM1 isoforms also have long (L) or short (S) cytoplasmic domains determined by the inclusion or exclusion of CEACAM1 exon 7 in the message. The CEACAM1L cytoplasmic domain has two ITIM motifs, which are unique to CEACAM1 among CEACAM family members.In one aspect, therapeutic anti-CEACAM1 antibodies or antigen-binding fragments thereof, including those described herein by their structural features, bind to the extracellular GF-CC' surface, which is the variable (V)-like Ig domain at the amino (N)-terminus of the CEACAM1 protein (N-domain), a domain common to all isoforms of CEACAM1, including CEACAM1 isoforms 1L, 1S, 3L, 3S, 4L, 4S, 3A1, 3AS, 3, 4C1, and 4C2. In some embodiments, the antibodies and antigen-binding fragments thereof provided bind to human CEACAM1. In some embodiments, the antibodies and antigen-binding fragments thereof provided bind to mammalian CEACAM1. The sequence of the full-length form of CEACAM1 (NCB1 Reference Sequence NP__p01703.2; UNIPROT ID P13688) is provided as SEQ ID NO:15 (signal sequence: residues 1-34 of SEQ ID NO:15; Ig-V N domain: residues 35-142 of SEQ ID NO:15. The mature form of CEACAM1 (without the signal sequence) is provided as SEQ ID NO:13.

[0137] As used herein, "binding" of an antibody or antigen-binding fragment thereof to CEACAM1, an epitope on CEACAM1, or, in certain embodiments described below, a particular residue on CEACAM1, includes selective interaction of the antibody or antigen-binding fragment thereof with CEACAM1. Binding therefore includes primary and secondary interactions, including, for example, hydrogen bonds, ionic interactions, and salt bridges, as well as hydrophilic and hydrophobic interactions.

[0138] In certain embodiments, the CEACAM1 antibody or antigen-binding fragment thereof described herein is -5 ~10 -12 mo1 / 1, 10 -6 ~10 -12 mol / l, 10 -7 ~10 -12 mol / l, 10 -8 ~10 -12 mol / l, 10 -9 ~10-12 mo1 / 1, 10 -10 ~10 -12 mo1 / 1, or 10 -11 ~10 -12 mo1 / 1K D In other embodiments, the CEACAM1 antibodies or antigen-binding fragments thereof described herein bind to CEACAM1 at 10 -5 ~10 -11 mo1 / 1, 10 -6 ~10 -11 mo1 / 1, 10 -7 ~10 -11 mo1 / 1, 10 -8 ~10 -11 mo1 / 1, 10 -9 ~10 -11 mo1 / 1, or 10 -10 ~10 -11 mo1 / 1K D In other embodiments, the CEACAM1 antibodies or antigen-binding fragments thereof described herein bind to CEACAM1 at 10 -5 ~10 -10 mo1 / 1, 10 -6 ~10 -10 mo1 / 1, 10 -7 ~10 -10 mo1 / 1, 10 -8 ~10 -10 mo1 / 1, or 10 -9 ~10 -10 mo1 / 1K D In other embodiments, the CEACAM1 antibodies or antigen-binding fragments thereof described herein bind to CEACAM1 at 10 -5 ~10 -8 mo1 / 1, 10 -6 ~10 -8 mo1 / 1, or 10 -7 ~10 -8 mo1 / 1K D and binds to CEACAM1.

[0139] The "specificity" of an antibody refers to the ability of an antibody or an antigen-binding fragment thereof, such as an anti-CEACAM1 antibody or an antigen-binding fragment thereof, to recognize a given epitope in CEACAM1 while having little or no detectable reactivity with non-CEACAM1 proteins or other parts of CEACAM1. Specificity can be relatively determined by competitive assays or by the epitope identification / characterization techniques described herein or their equivalents known in the art.

[0140] Epitopes can be formed from contiguous amino acids or from non-contiguous amino acids brought into close proximity by tertiary folding of a protein. Epitopes formed from contiguous amino acids are typically retained on exposure to denaturing solvents, while epitopes formed by tertiary folding are typically lost on treatment with denaturing solvents. An epitope typically comprises at least three, more usually at least five, about nine, or about eight to ten amino acids in a particular spatial conformation. An "epitope" comprises the unit of structure conventionally bound by an immunoglobulin VH / VL pair. An epitope defines the minimal binding site for an antibody or antigen-binding fragment thereof and is thus the target of the specificity of the antibody or antigen-binding fragment thereof. In the case of a single domain antibody, the epitope is the unit of structure bound by an independent variable domain.

[0141] In certain embodiments, contemplated antibodies or antigen-binding fragments specifically bind to the same epitope as antibody CP08H03 / Vk8 S29A. In another embodiment, contemplated antibodies or antigen-binding fragments bind to the same epitope as CP08H03 / CP08F05. Additional description of the creation of CP08 antibodies and the binding of antibodies CP08 / Vk8 S29A and CP08H03 / CP08F05 to the epitopes can be found in International Application No. PCT / US2019 / 065212, which is incorporated herein by reference in its entirety.

[0142] In one aspect, the antibody or antigen-binding fragment thereof, including the antibody described herein by its structural characteristics, specifically binds to at least a portion of the homophilic binding domain on CEACAM1 (i.e., the CEACAM1 protein portion involved in the formation of CEACAM1:CEACAM1 homodimer), thereby blocking CEACAM1 homophilic interaction.In certain embodiments, the provided antibody or antigen-binding fragment thereof is contained in the CC' and FG loops of CEACAM1, and specifically binds to one or more of the CEACAM1 residues, including the YQQN pocket (i.e., Y34, Q44, Q89, N97 of SEQ ID NO:13) in the CEACAM1:CEACAM1 dimer interface.See Huang et al., Nature. 2015 Jan 15;517(7534):386-90.

[0143] As used herein, a "blocking" or "antagonist" antibody is one that inhibits or reduces the biological activity of the antigen to which it binds. For example, in some embodiments, a CEACAM1 antagonist antibody or an antigen-binding fragment thereof binds to CEACAM1 and inhibits the activity of CEACAM1 and / or the binding of CEACAM1 to other CEACAM proteins or heterologous binding partners such as TIM-3. Inhibition of activity and inhibition of binding include partial inhibition. Methods for identifying CEACAM1 antibodies that block homophilic and heterophilic interactions of CEACAM1 are described herein and are known to those skilled in the art. For example, competing, cross-blocking, and cross-blocking antibodies can be identified using any suitable method known in the art, including competitive ELISA or BIACORE® assays, where the binding of the competing or cross-blocking antibody to human CEACAM1 blocks the binding of the antibody disclosed herein, or vice versa.

[0144] In certain embodiments, not all CDRs are directly involved in binding to antigen. In one embodiment, four of the six CDRs of CEACAM1 antibody or its antigen-binding fragment contact antigen. In one embodiment, five of the six CDRs of CEACAM1 antibody or its antigen-binding fragment contact antigen. In one embodiment, six of the six CDRs of CEACAM1 antibody or its antigen-binding fragment contact antigen. In one embodiment, CDR2H, CDR3H, CDR1L, CDR2L and CDR3L of CEACAM1 antibody or its antigen-binding fragment are directly involved in binding to antigen.

[0145] Despite the similarity of the N-domain among CEACAM family members, in some embodiments, the antibodies or antigen-binding fragments thereof provided herein, including those described herein by their structural features, are selective for CEACAM1. By selectively targeting CEACAM1, embodiments of the invention can, for example, avoid undesirable interference with the broad activation function of CEACAM3.

[0146] As used herein, the terms "selective" and "selectivity" refer to the preferential binding of an antibody or antigen-binding fragment thereof (i.e., a CEACAM1 antibody or antigen-binding fragment thereof) to a particular region, target, or peptide; typically a region or epitope in CEACAM1, as opposed to one or more other biological molecules, including other CEACAM family members.

[0147] In some embodiments, the CEACAM1 antibody or antigen-binding fragment thereof does not exhibit significant binding to CEACAM3, CEACAM5, CEACAM6, and / or CEACAM8. In some embodiments, the CEACAM1 antibody or antigen-binding fragment thereof does not exhibit detectable binding to CEACAM3, CEACAM5, CEACAM6, and / or CEACAM8. In some embodiments, a contemplated CEACAM1 antibody or antigen-binding fragment thereof binds to CEACAM1 with an affinity that is at least 10-fold, e.g., at least 100-fold, and at least 1000-fold, and up to 10,000-fold, or more, stronger than the affinity with which a contemplated CEACAM1 antibody or antigen-binding fragment thereof binds to another target or polypeptide.

[0148] In some cases, CEACAM1 antibody or antigen-binding fragment may not bind to CEACAM1. If both alleles of CEACAM1 gene are mutated or variant for A49, Q89, Q44, and / or Y34, no binding to CEACAM1 occurs. Such differences may be point mutations that change amino acids to A49V, Q89H, Q44L, and / or Y34C. One or all of the point mutations may occur in one or both alleles of CEACAM1 gene.

[0149] When an individual is heterozygous for one or more of these variant alleles, antibodies that bind to the wild-type allele may be less effective in treating a disease or disorder associated with CEACAM1 because the antibodies may exhibit reduced affinity or poor binding to the variant protein.

[0150] As used herein, "affinity" refers to the equilibrium dissociation constant (K D ) and is a measure of the binding strength between an antigenic determinant and an antigen-binding site on an antigen-binding protein, such as an antibody or antibody fragment thereof. D The smaller the value of K, the stronger the binding strength between the antigenic determinant and the antigen-binding molecule. Alternatively, affinity can be expressed as the affinity constant (KA ), where the constant is 1 / K D As will be apparent to one skilled in the art, affinity can be determined in a manner known per se depending on the particular antigen of interest.

[0151] In one aspect, anti-CEACAM1 antibody and its antigen-binding fragment, including the antibody described herein by its structural features, specifically binds to at least a portion of the binding site on CEACAM1 for one or more other members of the CEACAM family, thereby blocking CEACAM1 interaction with one or more other members of the CEACAM family.These CEACAM family members include, but are not limited to, CEACAM3, CEACAM5, CEACAM6, and CEACAM8 (Ramani et al, Anal. Biochem. Jan. 15, 2012; 420(2); 127-38; Scheffrahn et al, J. Immunol. May. 15, 2002; 168(10); 5139-46).

[0152] In one aspect, the anti-CEACAM1 antibody and its antigen-binding fragment, including the antibody described herein by its structural features, specifically binds to at least a portion of the binding site on CEACAM1 for a member of the TIM family, thereby blocking CEACAM1 interaction with the TIM family member.In some embodiments, the TIM family member is TIM-3.In some embodiments, the CEACAM1 antibody or its antigen-binding fragment specifically binds to one or more of the CEACAM1 residues Y34, G41, N42, Q44, Q89, S93, D94, V96, and / or N97 of SEQ ID NO:13, which have been shown to be involved in CEACAM1 binding to TIM-3 (Huang et al., Nature. 2015 Jan 15;517(7534):386-90).

[0153] To the extent that binding of an anti-CEACAM1 antibody to the GF-CC' face of CEACAM1 can interfere with or disrupt the interaction of the CEACAM1 polypeptide with factors that affect or mediate a CEACAM1-related disease or disorder, the antibody can provide a therapeutic benefit. The interaction of CEACAM1 with other factors associated with various pathologies is described herein for completeness.

[0154] In one aspect, anti-CEACAM1 antibodies and antigen-binding fragments thereof, including those described herein by their structural features, specifically bind to at least a portion of the binding site on CEACAM1 for a bacterial adhesive surface protein (adhesin), thereby blocking the interaction between CEACAM1 and the adhesin. In certain embodiments, the adhesin is expressed on the surface of a CEACAM1-binding pathogenic bacterium, including, but not limited to, Escherichia coli, specifically, diffuse adherent E. coli (DAEC), Neisseria gonorrhoeae, Neisseria meningitidis, commensal Neisseria, Streptococcus catarrhalis, Haemophilus influenzae, Haemophilus aegyptius, Helicobacter pylori, and / or Salmonella spp.

[0155] In one embodiment, the CEACAM1 antibody or antigen-binding fragment thereof disrupts the interaction between CEACAM1 and HopQ expressed on the surface of H. pylori. In one embodiment, the CEACAM1 antibody or antigen-binding fragment specifically binds to one or more of CEACAM1 residues F29, Y34, N42, Q89, and N97, which are predicted to be involved in CEACAM1 binding to HopQ.

[0156] In another embodiment, the CEACAM1 antibody or antigen-binding fragment thereof disrupts the interaction between CEACAM1 and opacity-associated (Opa) adhesin proteins expressed on the surface of Neisseria species, including but not limited to Opa52, Opa65, Opa70, Opa72, Opa73, Opa74, and Opa75. In one embodiment, the CEACAM1 antibody or antigen-binding fragment specifically binds to one or more of CEACAM1 residues Q44 and A49, which are predicted to be involved in CEACAM1 binding to Neisserial Opa proteins.

[0157] In another embodiment, the CEACAM1 antibody or antigen-binding fragment thereof disrupts the interaction between CEACAM1 and the Opa-like protein O1pA expressed on the surface of Moraxella spp.

[0158] In one embodiment, the CEACAM1 antibody or antigen-binding fragment thereof disrupts the interaction between CEACAM1 and H. influenzae OMP P1. In one embodiment, the CEACAM1 antibody or antigen-binding fragment specifically binds to one or more of CEACAM1 residues Q44 and A49, which are predicted to be involved in CEACAM1 binding to H. influenzae OMP P1.

[0159] In another embodiment, the CEACAM1 antibody or antigen-binding fragment thereof disrupts the interaction between CEACAM1 and Haemophilus aegyptius OMP P1. In one embodiment, the CEACAM1 antibody or antigen-binding fragment specifically binds to CEACAM1 residue F29, which is predicted to be involved in CEACAM1 binding to Haemophilus aegyptius OMP P1.

[0160] In another embodiment, the CEACAM1 antibody or antigen-binding fragment thereof disrupts the interaction between CEACAM1 and Candida albicans.

[0161] In another embodiment, the CEACAM1 antibody or antigen-binding fragment thereof disrupts the interaction between CEACAM1 and influenza viruses, including but not limited to H5N1.

[0162] In another embodiment, the CEACAM1 antibody or antigen-binding fragment thereof described herein inhibits binding of CEACAM1 to a nematode. In one embodiment, the nematode is B. bancrofti.

[0163] nucleic acid In some embodiments, nucleic acids encoding CEACAM1 antibodies and antigen-binding fragments thereof, as well as vectors, host cells, and expression systems containing such nucleic acids, can be used to prepare or deliver anti-CEACAM1 antibodies.

[0164] The nucleic acid encoding CEACAM1 antibody or its antigen-binding fragment may be, for example, DNA, cDNA, RNA, synthetically produced DNA or RNA, or recombinantly produced chimeric nucleic acid molecule comprising any of these polynucleotides alone or in combination.For example, the nucleic acid may comprise an expression vector comprising the polynucleotide sequence encoding the CEACAM1 antibody or its antigen-binding fragment described herein, operably linked to an expression control sequence suitable for expression in eukaryotic and / or prokaryotic host cells.

[0165] Antibody preparation and expression system The antibodies or antigen-binding fragments thereof described herein are typically produced by recombinant expression. Nucleic acids encoding the light chain variable region and the heavy chain variable region, optionally linked to a constant region, are inserted into an expression vector. The light and heavy chains can be cloned into the same or different expression vectors. The DNA segments encoding the immunoglobulin chains are operably linked to control sequences in the expression vector(s) that ensure the expression of the immunoglobulin polypeptide. Expression control sequences include, but are not limited to, promoters (e.g., naturally associated or heterologous promoters), signal sequences, enhancer elements, and transcription termination sequences. Preferably, the expression control sequences are eukaryotic promoter systems in vectors capable of transforming or transfecting eukaryotic host cells. Once the vector is incorporated into a suitable host cell, the host cell is maintained under conditions suitable for high-level expression of the nucleotide sequence and collection and purification of the cross-reactive antibodies. Expression systems for antibodies are described, for example, in PCT / US2019 / 065212, which is incorporated herein by reference.

[0166] Genomic DNA extraction and genotyping assays The methods described herein include genotyping an individual for the individual's CEACAM1 genotype. Such genotyping is typically performed on a biological sample containing genomic DNA from the subject. Several suitable methods are available in the art, including, but not limited to, DNA sequencing, allele-specific hybridization, PCR, allele-specific amplification (ASA) (WO93 / 22456), allele-specific hybridization, single-base extension (U.S. Pat. No. 4,656,127), ARMS-PCR, Taqman (U.S. Pat. Nos. 4,683,202; 4,683,195; and 4,965,188), oligoligation assay, single-strand conformation analysis ((SSCP) Orita et al PNAS 86 2766-2770 (1989)), gene bit analysis (WO92 / 15712) and RFLP direct sequencing, mass spectrometry (MALDI-TOF), and DNA array. In some embodiments, the biological sample includes cells or tissues or peripheral blood or body fluids. Exemplary biological samples include, but are not limited to, biopsies, tumor samples, biological fluid samples; blood; serum; plasma; urine; semen; mucus; tissue biopsies; organ biopsies; synovial fluid; bile; cerebrospinal fluid; mucosal secretions; exudates; sweat; saliva; and / or tissue samples. The term also includes mixtures of the above-mentioned samples. The term "test sample" also includes untreated or pretreated (or pre-processed) biological samples. In some embodiments of any of the aspects, the test sample may include cells from a subject. In some embodiments of any of the aspects, the test sample may be a body fluid, a buccal swab, or a tumor sample.

[0167] The above methods may require amplification of a DNA sample from the subject, which may be performed by techniques known in the art, such as PCR (see PCR Technology: Principles and Applications for DNA Amplification (ed. H. A. Erlich, Freeman Press, NY 1992; PCR Protocols: A Guide to methods and Applications (eds. Innis et al., Academic press, San Diego, Calif. 1990); Mattila et al., Nucleic Acids Res. 19 4967 (1991); Eckert et al., PCR Methods and Applications 117 (1991), and U.S. Pat. No. 4,683,202. Other suitable amplification methods include ligase chain reaction (LCR) (Wu et al., Genomics 4 560 (1989); Landegran et al., Science 241 1077 (1988)), transcription amplification (Kwoh et al., Proc Natl Acad Sci USA 86 1173 (1989)), self-sustained sequence replication (Guatelli et al., Proc Natl Acad Sci USA 87 1874 (1990)), and nucleic acid-based sequence amplification (NASBA). Both of the latter two methods involve an isothermal reaction based on isothermal transcription that produces both single-stranded RNA and double-stranded DNA as amplification products, in a ratio of 30:1 or 100:1, respectively.

[0168] Exemplary methods for processing test samples include, but are not limited to, centrifugation, filtration, sonication, homogenization, heating, freezing and thawing, and combinations thereof. In some embodiments, the test sample may be a frozen test sample, such as, for example, frozen tissue. The frozen sample may be thawed before using the methods or assays described herein. After thawing, the frozen sample may be centrifuged before being subjected to the methods or assays described herein. In some embodiments, the test sample is a clarified test sample, for example, prepared by centrifugation and collection of the supernatant. In some embodiments, the test sample may be a pre-processed test sample, such as, for example, the supernatant or filtrate obtained from a process selected from centrifugation, filtration, thawing, purification, and any combination thereof. In some embodiments, the test sample may be treated with chemical and / or biological reagents. Chemical and / or biological reagents may be used to protect and / or maintain the stability of the sample, including the biological molecules (e.g., nucleic acids and proteins) present therein, during storage or processing. One exemplary reagent is a protease inhibitor, which is generally used to protect or maintain the stability of proteins during processing. Those skilled in the art are familiar with methods and processes suitable for pre-treatment of biological samples required for determining the levels of expression products described herein.

[0169] In some embodiments of any of the aspects, the methods and assays described herein may further include obtaining or having obtained a test sample from a subject.

[0170] Nucleic acid samples obtained from a subject's cancerous and non-cancerous cells that can be used in the methods described herein to determine a subject's genotype can be prepared by means well known in the art.

[0171] While the sample most likely to be used for subject genotyping is a blood sample, in some cases, taking nucleic acid samples from subject can be achieved, for example, by surgery or aspiration.In surgical procedures to remove cancerous tissue, surgeons often remove the non-cancerous tissue and / or cell samples of the same tissue type from cancer patients for comparison.Nucleic acid samples can be isolated from such tissues of subject for use in the methods described herein.

[0172] Protein extraction and antibody binding assays / affinity determination Where applicable, the amount of CEACAM1 expressed by a subject or in or around a cancer or infected tissue of a subject can be determined by methods known in the art, including, for example, ELISA, Western blot, immunohistochemistry, or other immunoassays. Such immunoassays can be performed using antibodies that bind to the GF-CC' face of CEACAM1, or in other embodiments, antibodies that bind to other epitopes. Mass spectrometry can also be used. Standard techniques known in the art for ELISA are described in "Methods in Immunodiagnosis", 2nd Edition, Rose and Bigazzi, eds. John Wiley & Sons, 1980; and Oellerich, M. 1984, J. Clin. Chem. Clin. Biochem. 22:895-904. These references are incorporated herein by reference in their entirety.

[0173] A level that is less than the reference level can be at least about 10% lower, at least about 20% lower, at least about 50% lower, at least about 60% lower, at least about 80% lower, at least about 90% lower, or lower than the reference level. In some embodiments, a level that is less than the reference level can be a statistically significantly lower level compared to the reference level.

[0174] A level above the reference level can be at least about 10% greater, at least about 20% greater, at least about 50% greater, at least about 60% greater, at least about 80% greater, at least about 90% greater, at least about 100% greater, at least about 200% greater, at least about 300% greater, at least about 500% greater, or greater than the reference level. In some embodiments, a level above the reference level can be a level that is statistically significantly greater than the reference level.

[0175] In some embodiments, if a subject is found to be heterozygous for one or more allelic variants of CEACAM1, including A49V, Q89H, Q44L, and / or Y34C, the methods described herein may further comprise assaying a sample containing CEACAM1 from the subject to determine the extent to which the subject's CEACAM1 is bound by an agent comprising an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1. The amount and / or dosing regimen for such an agent may be adjusted as a function of the degree of binding of the agent to the subject's CEACAM1. That is, if an antibody agent is found to bind to the subject's CEACAM1 protein, but with, for example, a reduced affinity, the dosage and / or frequency of dosing of the antibody agent received by the subject may be increased to improve efficacy. Such an increase in dosage may be proportional to, for example, the degree of reduced affinity determined.

[0176] Methods for measuring antibody binding are known to those skilled in the art. Such methods include, for example, ELISA (enzyme-linked immunosorbent assay), Western blot, immunoprecipitation, and immunofluorescence assay. Antibody dissociation constants and affinities can be determined, for example, by surface plasmon resonance-based assays (such as the BIAcore assay described in PCT Publication No. WO2005 / 012359); Forte Bio Octet™ analysis, enzyme-linked immunosorbent assay (ELISA); and competitive assays (e.g., RIA). The antibodies or antigen-binding fragments thereof described herein have an affinity (as described above, suitably, e.g., K ) of at least 1000-fold, 10000-fold, or greater than the affinity with which the amino acid sequence or polypeptide binds to another given polypeptide. D A molecule is said to be "specific" or "specifically binds" or "selectively binds" to a first target or antigen when it binds to the first antigen with a specific binding affinity (expressed as a specific affinity value) relative to a second target or antigen. Generally, a molecule that "specifically binds," "selectively binds," or is "specific" for a given target is said to be "specific" or "binds specifically" or "selectively binds" to a given target with a specific binding affinity of at least 10 -6 M (1000 nM) or less, e.g., 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M or lower K DIt is believed that the polypeptides bind at a specific binding site. Specific binding can be affected, for example, by the affinity and avidity of the polypeptide agent as well as the concentration of the polypeptide agent. One skilled in the art can use any method, such as titrating the polypeptide agent in a suitable binding assay, to determine the appropriate conditions under which the polypeptide agents described herein selectively bind to the target. ELISA can be suitable, for example, via competitive ELISA, to obtain a measure of antibody affinity. Other techniques for determining the affinity of a given antibody (e.g., an antibody that binds to the GF-CC' face of CEACAM1, e.g., a CP08H03 / Vk8 S29A or CP08H03 / CP08F05-based antibody agent) are known in the art. Non-limiting examples include surface plasmon resonance (e.g., via a Biacore™ assay) and ionolayer interference (BLI), e.g., via an Octet™ assay (Forte Bio). In such assays, the affinity of an antibody to CEACAM1 in a sample from a subject can be determined.

[0177] Generally, antibodies have a K greater than 1 micromolar. D In other embodiments, if the antibody is shown to bind to CEACAM1 in a subject at 10 -7 K is higher than M D If the affinity is in the range of 100 nM to 1 nM, then increasing the dose and / or frequency of administration may be adjusted to increase the dose in proportion to the degree of difference in affinity for binding to wild-type CEACAM by the same antibody agent. The CEACAM1 antibodies or antigen-binding fragments thereof described herein may be administered at 10 -6 ~10 -12 mol / l, 10 -7 ~10 -12 mol / l, 10 -8 ~10 -12 mol / l, 10 -9 ~10 -12 mo1 / 1, 10-10 ~10 -12 mo1 / 1, or 10 -11 ~10 -12 mo1 / 1K D and binds to CEACAM1.

[0178] Methods for modulating CEACAM1 activity In one aspect, the anti-CEACAM1 antibodies and antigen-binding fragments thereof described herein are useful in decreasing the interaction between CEACAM1 and other members of the CEACAM family, including, but not limited to, CEACAM1, CEACAM3, CEACAM5, CEACAM6, and CEACAM8. In some embodiments, the antibodies or antigen-binding fragments thereof disrupt homophilic interactions between CEACAM1 monomers.

[0179] In another aspect, the anti-CEACAM1 antibodies and antigen-binding fragments thereof described herein are useful in decreasing the interaction between CEACAM1 and members of the TIM family, including TIM-3. In some embodiments, the antibodies or antigen-binding fragments thereof disrupt the heterophilic interaction between CEACAM1 and TIM-3. Disruption of the interaction between CEACAM1 and TIM-3 by using the antibodies and antigen-binding fragments thereof contemplated by the present invention can reverse the CEACAM1 inhibitory function while maintaining the TIM-3 activating function.

[0180] Inhibition of CEACAM1 may be useful in reducing immune suppression, such as T cell tolerance. Immune suppression may be mediated by immune inhibitory receptors expressed on the surface of immune cells and their interaction with their ligands. For example, cytotoxic CD8 T cells may enter a state of "functional exhaustion" or "refractory" in which they express inhibitory receptors that block antigen-specific responses such as proliferation and cytokine production. Thus, by inhibiting the activity and / or expression of such inhibitory receptors, immune responses against cancer or tumors that are suppressed, inhibited, or refractory may be enhanced or relieved. Such enhancement or inhibition reversal of immune response may lead to improved capacity or receptivity for T cell activity, responsiveness, and / or activation.

[0181] Methods for measuring T cell activity and thereby measuring the relative reduction or absence of tolerance or activity are known in the art. Thus, in some embodiments, one or more of the parameters or markers of T cell tolerance can be assayed to determine whether the anti-CEACAM1 antibody or antigen-binding fragment thereof disclosed herein inhibits CEACAM1 activity. As an example, a reduction in the level of a factor associated with tolerance can indicate tolerance inhibition, including, but not limited to, a reduction in the level of IL-2. Other cytokines and factors associated with or involved in tolerance are known in the art. Reduction of T cell tolerance can also be assessed by examining tumor-infiltrating lymphocytes or T lymphocytes in lymph nodes draining established tumors. Such T cells show characteristics of exhaustion, for example, through the expression of cell surface molecules such as PD1, TIM-3, or LAG-3, and the reduced secretion of cytokines such as interferon-γ. Thus, evidence that T cell tolerance is reduced in the presence of a CEACAM1 antibody or antigen-binding fragment thereof includes, for example, an increase in the amount of T cells with (a) antigen specificity for a tumor-associated antigen (e.g., as determined by a major histocompatibility complex class I or class II tetramer containing a tumor-associated peptide) and (b) high levels of secretion of cytolytic effector molecules such as interferon-γ and granzyme-B compared to that seen in the absence of the antibody agent.

[0182] Treatment Method In one aspect, described herein is a method for determining whether a disease or disorder of a given subject is likely to respond to a given CEACAM1 antibody agent.In another aspect, described herein is a method for treating a subject whose disease or disorder is found to be likely to respond to a CEACAM antibody agent with such an agent.In such a method, a therapeutically effective amount of an antibody or antigen-binding portion thereof is administered to a mammal in need thereof.The antibodies or antigen-binding portions thereof described herein are particularly useful for administration to humans, but they can also be administered to other mammals.A "therapeutically effective amount" is an amount of an antibody agent described herein that is effective in providing a desired therapeutic effect when administered to a mammal.

[0183] In some aspects, the antibody or antigen-binding fragment thereof binds to CEACAM1 expressed by exhausted T cells or natural killer (NK) cells, thereby restoring T cell and NK cell activity and causing an increased anti-tumor response. In other aspects, the antibody or antigen-binding fragment thereof binds to CEACAM1 expressed by tumor cells, thereby inhibiting tumor cell metastasis and the formation of cancer stem cell niches. In yet another aspect, the antibody or antigen-binding fragment thereof binds to CEACAM1 expressed by macrophages associated with fibrosis in the tumor environment, thereby inhibiting fibrosis. In another aspect, the antibody or antigen-binding fragment thereof binds to CEACAM1 expressed by other stromal cells in the tumor microenvironment, such as vascular endothelial cells, thereby inhibiting angiogenesis.

[0184] Therefore, the present specification also provides a method for treating a subject with cancer or tumor and / or reducing tumor growth, comprising administering an effective amount of the CEACAM1 antibody agent described herein.In this context, "reducing" includes inhibiting and / or reversing, and can refer to, for example, the symptoms of the disorder being treated, the presence or size of metastasis or micrometastasis, the size of primary tumor, or the presence or size of dormant tumor.

[0185] By "subject" is intended a mammal, including but not limited to a human or a non-human mammal such as a cow, horse, dog, sheep, or cat. As used herein, an individual and a patient are also subjects.

[0186] The terms "treat", "treated", "treating" or "treatment" as used herein refer to therapeutic treatment, the purpose of which is to delay (alleviate) an undesirable physiological condition, disorder or disease, or to obtain a beneficial or desirable clinical outcome. For the purposes of this disclosure, beneficial or desirable clinical outcomes include, but are not limited to, alleviation of symptoms; reduction in the extent of a condition, disorder or disease; stabilization of the condition, disorder or disease state (i.e., no progression); slowing or delaying the onset of the progression of a condition, disorder or disease; improvement of the condition, disorder or disease state, and detectable or undetectable remission (whether partial or total), or fullness or amelioration of a condition, disorder or disease. Treatment includes eliciting a clinically meaningful response without excessive levels of side effects. Treatment also includes extending survival compared to the expected survival in the absence of treatment. The terms "prevent", "prevention", and the like refer to acting prior to the onset of an obvious disease or disorder, preventing a disease or disorder from developing, or minimizing the extent of a disease or disorder or slowing its developmental process.

[0187] The embodiments described herein can be used to treat metastasis, which refers to the spread of cancer from its primary site to other locations in the body. Cancer cells can break away from the primary tumor, invade lymphatic and blood vessels, circulate through the bloodstream, and grow in distant foci in normal tissues (to which they metastasize) elsewhere in the body. Metastasis can be local or distant. Metastasis is a continuous process that requires tumor cells to break away from the primary tumor, travel through the bloodstream, and reach distant sites. At the new site, the cells can establish a blood supply, grow, and form life-threatening masses. Both stimulatory and inhibitory molecular pathways within tumor cells regulate this behavior, and interactions between tumor cells and host cells at distant sites are also prominent. Metastasis is most often detected through the use of magnetic resonance imaging (MRI) scans, computed tomography (CT) scans, blood and platelet counts, liver function tests, chest X-rays, and bone scans, alone or in combination, in addition to monitoring for certain symptoms.

[0188] Also contemplated is a method of reducing cancer stemness, comprising administering the CEACAM1 antibody agent described herein. Cancer stemness can refer to the ability of cells to self-renew and give rise to additional phenotypically distinct cell types. Cancer stem cells (CSCs) are cancer cells that exhibit stem cell-like properties. CSCs often exhibit at least one characteristic of cancer and have the ability to give rise to at least one additional, phenotypically distinct cell type. In addition, cancer stem cells have the ability to perform both asymmetric and symmetric replication. It will be understood that cancer stem cells can arise from differentiated cancer cells that have acquired stem cell traits and / or stem cells that have acquired phenotypes associated with cancer cells. Alternatively, cancer stem cells can reconstitute non-stromal cell types within tumors.

[0189] CEACAM1 is expressed by many tumor types and CEACAM1 may control or be involved in the regulation of tumor growth and metastatic behavior. In another embodiment, CEACAM1 inhibition reduces tumor growth and metastasis.

[0190] CEACAM1 expression on a subset of macrophages is associated with fibrosis during carcinogenesis. In a further aspect, CEACAM1 inhibition reduces tumor-associated fibrosis.

[0191] Cancers that can be treated by the compositions and methods contemplated by the present invention include tumors that are not vascularized or are not yet substantially vascularized, and tumors that are vascularized.Cancers can include non-solid tumors (e.g., blood tumors such as leukemia and lymphoma) or can include solid tumors.Types of cancers to be treated include, but are not limited to, sarcomas, carcinomas, and leukemia, lymphomas, and myelomas, blastomas, and brain and spinal cancer.Also included are adult tumors / cancers and pediatric tumors / cancers. More specific examples of such cancers include: basal cell carcinoma, biliary tract cancer; bladder cancer; bone cancer; brain and CNS cancer; breast cancer; cancer of the peritoneum; cervical cancer; choriocarcinoma; colon and rectal cancer; connective tissue cancer; cancer of the digestive system; endometrial cancer; esophageal cancer; eye cancer; head and neck cancer; gastric cancer (including gastrointestinal cancer); glioblastoma; hepatoma; hepatoma; intraepithelial neoplasia; renal or kidney cancer; laryngeal cancer; leukemia; liver cancer; lung cancer (e.g., small cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, and squamous cell carcinoma of the lung); hominis; Lymphomas, including Hodgkin's lymphoma and non-Hodgkin's lymphoma; melanoma; myeloma; neuroblastoma; oral cancer (e.g., lip, tongue, mouth, and throat); ovarian cancer; pancreatic cancer; prostate cancer; retinoblastoma; rhabdomyosarcoma; rectal cancer; cancer of the respiratory system; salivary gland cancer; sarcoma; skin cancer; squamous cell carcinoma; stomach cancer; testicular cancer; thyroid cancer; uterine or endometrial cancer; cancer of the urinary system; vulvar cancer; and other cancers and sarcomas; and B-cell lymphomas (low-grade / follicular non-Hodgkin's lymphoma (NHL); small lymphocytic (SL) The cancers include, but are not limited to, NHL; intermediate grade / follicular NEIL; intermediate grade diffuse NHL; high grade immunoblastic NEIL; high grade lymphoblastic NEIL; high grade small noncleaved cell NHL; bulky disease NEIL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's macroglobulinemia; chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); hairy cell leukemia; chronic myeloblastic leukemia; and post-transplant lymphoproliferative disorder (PTLD), as well as abnormal blood vessel proliferation associated with nematoses, edema (such as those associated with brain tumors), and Meigs syndrome. Patients may have multiple types of cancer.

[0192] The effectiveness of the cancer treatment method, including the therapeutic preparation of the antibody and its antigen-binding fragment described herein, can be measured by various endpoints commonly used in evaluating cancer treatment, including, but not limited to, tumor regression, tumor weight or size shrinkage, time to progression, survival time, progression-free survival time, overall response rate, duration of response, and quality of life.In the case of cancer, a therapeutically effective amount of CEACAM1 antibody agent can reduce the number of cancer cells; reduce tumor size; inhibit (i.e., delay to a certain extent, preferably stop) cancer cell invasion into peripheral organs; inhibit (i.e., delay to a certain extent, preferably stop) tumor metastasis; inhibit tumor growth to a certain extent; and / or alleviate to a certain extent one or more symptoms associated with the disorder.In the case of a patient having multiple types of cancer, a therapeutically effective amount of CEACAM1 antibody agent is an amount that is effective in treating at least one of the cancers.The CEACAM1 antibody agent can be cytostatic and / or cytotoxic to the extent that it acts to inhibit the growth and / or kill existing cancer cells. For cancer therapy, in vivo efficacy may be measured, for example, by assessing survival time, progression-free survival (PFS) time, response rate (RR), duration of response, and / or quality of life.

[0193] Immune checkpoint proteins interact with specific ligands that signal T cells to turn off or inhibit T cell function. Cancer cells may suppress anti-cancer immune responses by controlling the function of T cells entering the tumor microenvironment by expressing high levels of checkpoint proteins on their surface. The immune checkpoint protein programmed cell death-1 (PD-1) is a key immune checkpoint receptor expressed by activated T and B cells and mediates immune suppression. PD-1 is a member of the CD28 receptor family that includes CD28, CTLA-4, ICOS, PD-1, and BTLA. Two cell surface glycoprotein ligands for PD-1 have been identified, programmed cell death ligand-1 (PD-L1) and programmed cell death ligand-2 (PD-L2), which are expressed on antigen-presenting cells and in many human cancers and have been shown to downregulate T cell activation and cytokine secretion upon binding to PD-1 (Freeman et al., 2000; Latchman et al., 2001). Inhibition of PD-1 / PD-L1 interaction can promote potent antitumor activity. Examples of PD-1 inhibitors include, but are not limited to, pembrolizumab (MK-3475), nivolumab (MDX-1106), cemiplimab-rwlc (REGN2810), pidilizumab (CT-011), spartalizumab (PDR001), tislelizumab (BGB-A317), PF-06801591, AK105, BCD-100, BI754091, JS001, LZM009, MEDI0680, MGA012, Sym021, and TSR-042. Examples of PD-L1 inhibitors include, but are not limited to, atezolizumab (MPDL3280A), durvalumab (MEDI4736), avelumab (MSB0010718C), BGB-A333, CK-301, CS1001, FAZ053, KN035, MDX-1105, MSB2311, and SHR-1316.

[0194] However, among cancer patients who undergo checkpoint inhibitor therapy, there is a significant population that (1) does not respond to this type of treatment (natural or primary resistance) or (2) responds initially but eventually develops disease progression (secondary or acquired resistance). Resistant cancers may also be referred to as resistant cancers. As shown in the PD-1 / CEACAM1 data from PCT / US2019 / 065212, the entirety of which is incorporated herein by reference, tumor-associated cells isolated from patients with acquired resistance to PD-1 / PD-L1 inhibitors upregulate CEACAM1 expression compared to tumor-associated cells isolated from naive patients who had not been exposed to PD-1 inhibitors. When CEACAM1 is expressed in the context of acquired resistance, cells with CEACAM1 are more likely to be effector memory cells rather than central memory cells, which is consistent with the reduced anti-cancer response in resistant patients.

[0195] Thus, among the methods described herein for using CEACAM1 antibody agents, include the use of such agents for the treatment of patients with resistance to checkpoint inhibitors, such as inhibitors of PD-1, PD-L1, and / or CTLA-4. In some embodiments, the CEACAM1 antibody used in the treatment of patients with resistance to inhibitors of PD-1, PD-L1, and / or CTLA-4 is an agent that includes CP08H03 / Vk8 S29A or CP08H03 / CP08F05 or an antigen-binding domain(s) thereof. In some embodiments, the resistance is natural resistance or primary resistance. In some embodiments, the resistance is secondary resistance or acquired resistance. In some embodiments, the administered CEACAM1 antibody agent reverses T cell exhaustion in patients with resistance to checkpoint inhibitor treatment. Any cancer that exhibits resistance to PD-1, PDL-1, and / or CTLA-4 inhibitors is suitable for treatment with such methods. In some embodiments, the CEACAM1 antibody agent is administered to a patient who has not previously received checkpoint inhibitor treatment.

[0196] In another aspect, the method includes the use of a CEACAM1 antibody agent in the treatment of patients resistant to therapy with other checkpoint inhibitors, including but not limited to inhibitors of PD-L2, B7-H3, B7-H4, BTLA, HVEM, GALS, LAG3, TIM-3, VISTA, KIR, 2B4 (belonging to the CD2 molecule family and expressed on all NK cells, y6 cells, and memory CD8+ (4) T cells), CD160 (also called BY55), CGEN-15049, CHK1 and CHK2 kinases, A2aR, and various B-7 family ligands (including but not limited to B7-1, B7-2, B7-DC, B7-H1, B7-H2, B7-H3, B7-H4, B7-H5, B7-H6, and B7-H7).

[0197] In another aspect, the method comprises using a CEACAM1 antibody agent to treat a subject in need of reducing and / or preventing colonization of mammalian epithelia by Candida albicans and / or bacteria expressing bacterial adhesins (including, but not limited to, Escherichia coli, specifically, diffuse adherent E. coli (DAEC), Neisseria gonorrhoeae, Neisseria meningitidis, commensal Neisseria, Streptococcus catarrhalis, Haemophilus influenzae, Haemophilus aegyptius, Helicobacter pylori, and / or Salmonella spp.). In another aspect, the method comprises using a CEACAM1 antibody agent to reduce influenza virus replication and / or to reduce release of proinflammatory cytokines or chemokines associated with infection with influenza virus. In some embodiments, the influenza virus is H5N1. In another aspect, the method comprises using a CEACAM1 antibody agent to treat a subject in need of reducing and / or preventing infection with filarial nematodes such as Wuchereria bancrofti. In another aspect, the method comprises using a CEACAM1 antibody agent to treat a subject in need of reducing and / or preventing the development of lymphedema and / or water masses associated with infection with a filarial nematode, such as W. bancrofti. In one embodiment, the method comprises using a CEACAM1 antibody agent to reduce invasion of the lymphatic system of the subject by filarial nematodes in a subject in need thereof. In one embodiment, the filarial nematode is W. bancrofti. The subject may be infected by more than one of bacteria expressing a bacterial adhesin, Candida albicans, influenza virus, and / or filarial nematodes.

[0198] In another embodiment, the method comprises using a CEACAM1 antibody agent in a subject in need thereof to reduce invasion of the subject's lymphatic system by cancer cells.

[0199] Pharmaceutical Compositions If the genotyping step indicates that the subject's disease or disorder can be treated with an anti-CEACAM1 antibody agent (e.g., an agent that binds to the GF-CC' face of CEACAM1, including, but not limited to, a CP08H03 / Vk8S29A or CP08H03 / CP08F05-based antibody agent), the antibody agent may comprise a therapeutically effective amount of a CEACAM1 antibody or antigen-binding fragment thereof described herein formulated together with one or more pharma- ceutically acceptable excipients.

[0200] The dosage of active agent(s) may vary depending on the reason for use, the individual subject, and the mode of administration. The dosage may be adjusted based on the subject's weight, the subject's age and health, and the tolerance to the compound(s) or composition. For example, for an antibody or its antigen-binding fragment, depending on the disease, this may require 0.1, 1.0, 3.0, 6.0, or 10.0 mg / Kg. For an IgG (two binding sites) with a molecular mass of 150,000 g / mol, these dosages correspond to approximately 18 nM, 180 nM, 540 nM, 1.08 μM, and 1.8 μM of binding sites per 5L blood volume.

[0201] The active agent and excipient(s) can be formulated into compositions and dosage forms according to methods known in the art.The pharmaceutical compositions of the present invention can be specially formulated in solid or liquid form, including those suitable for parenteral administration, for example, by subcutaneous injection, intratumoral injection, intramuscular injection, or intravenous injection, for example, as a sterile solution or suspension.

[0202] Therapeutic compositions comprising an antibody or antigen-binding fragment thereof that binds to CEACAM1 can be formulated with one or more pharma- ceutically acceptable excipients, which can be liquid or solid fillers, diluents, carriers, manufacturing aids (e.g., lubricants, magnesium talc, calcium or zinc stearate, or steric acid), bulking agents, salts, surfactants, and / or preservatives, solvents, or encapsulating materials involved in carrying or transporting a therapeutic compound for administration to a subject. Some examples of materials that can serve as pharma- ceutically acceptable excipients include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; gelatin; talc; waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as ethylene glycol and propylene glycol; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers; water; isotonic saline; pH buffers; and other non-toxic compatible substances used in pharmaceutical formulations.

[0203] Bulking agents are compounds that impart mass to a pharmaceutical formulation and contribute to the physical structure of the formulation in lyophilized form. Suitable bulking agents include, for example, mannitol, glycine, polyethylene glycol, and sorbitol.

[0204] The use of a surfactant may reduce aggregation of the reconstituted protein and / or reduce the formation of particulates in the reconstituted formulation. The amount of surfactant added is such that it reduces aggregation of the reconstituted protein and minimizes the formation of particulates after reconstitution. Suitable surfactants include polysorbates (e.g., polysorbate 20 or 80); poloxamers (e.g., poloxamer 188); Triton; sodium dodecyl sulfate (SDS); sodium laurel sulfate; sodium octyl glycoside; lauryl sulfobetaine, myristyl sulfobetaine, linoleyl sulfobetaine, or stearyl sulfobetaine; lauryl sarcosine, myristyl sarcosine, linoleyl sarcosine, or stearyl sarcosine; linoleyl betaine, myristyl betaine, or cetyl betaine; lauroamidopropyl betaine, cocamide protease, or glyceryl stearate; ... myristamidopropyl betaine, linoleamidopropyl betaine, myristamidopropyl betaine, palmidopropyl betaine, or isostearamidopropyl betaine (e.g., lauroamidopropyl); myristamidopropyl dimethylamine, palmidopropyl dimethylamine, or isostearamidopropyl dimethylamine; sodium methyl cocoyl taurate or disodium methyl oleyl taurate; and polyethyl glycol, polypropyl glycol, and copolymers of ethylene and propylene glycol (e.g., Pluronics, PF68, etc.).

[0205] Preservatives may be used in the formulations of the invention. Preservatives suitable for use in the formulation include octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride (a mixture of alkylbenzyldimethylammonium chlorides in which the alkyl group is a long-chain compound), and benzethonium chloride. Other types of preservatives include aromatic alcohols such as phenol, butyl, and benzyl alcohol, alkylparabens such as methylparaben or propylparaben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol. Other suitable pharmaceutical excipients can be found in standard pharmaceutical textbooks, such as "Remington's Pharmaceutical Sciences", The Science and Practice of Pharmacy, 19th Ed. Mack Publishing Company, Easton, Pa., (1995).

[0206] The composition comprising the antibody or antigen-binding fragment thereof and a pharma- ceutically acceptable carrier may contain the CEACAM1 antibody or antigen-binding portion thereof at various concentrations. For example, the composition may contain the antibody or antigen-binding fragment thereof at 10 mg / ml to 200 mg / ml, 25 mg / ml to 130 mg / ml, 50 mg / ml to 125 mg / ml, 75 mg / ml to 110 mg / ml, or 80 mg / ml to 100 mg / ml. The composition may also contain the antibody or antigen-binding fragment thereof at about 10 mg / ml, 20 mg / ml, 30 mg / ml, 40 mg / ml, 50 mg / ml, 60 mg / ml, 70 mg / ml, 80 mg / ml, 90 mg / ml, 100 mg / ml, 110 mg / ml, 120 mg / ml, 130 mg / ml, 140 mg / ml, or 150 mg / ml.

[0207] In some embodiments, a composition comprising an antibody or antigen-binding fragment thereof and a pharma- ceutically acceptable carrier is lyophilized and provided as a composition for reconstitution prior to administration.

[0208] Method of administration In some embodiments, the method described herein relates to treating subjects with or diagnosed as having CEACAM1-related disease or disorder with anti-CEACAM1 antibody agent.Subjects with CEACAM1-related disease or disorder can be identified using current methods for diagnosing CEACAM1-related disease or disorder, including but not limited to T cell exhaustion, changes in the infiltration pattern of immune cells in microenvironment, T cell refractoriness, low levels of T cells, increased regulatory T cells, and / or increased expression of CEACAM1.Symptoms and / or complications of CEACAM1-related disease or disorder that characterize and diagnose such conditions are well known in the art, and include but are not limited to fatigue, weight loss, abnormal swelling and / or mass, any pain of new or unknown origin, cough, hoarseness, abnormal bleeding, changes in bowel or bladder habits, fever, night sweats, headache, vision or hearing problems, or oral changes such as pain, bleeding, pain, or numbness.

[0209] The compositions and methods described herein can be administered to subjects who have or have been diagnosed as having a CEACAM1-associated disease or disorder. In some embodiments, the methods described herein include administering to a subject an effective amount of a composition described herein, such as, for example, a CEACAM1 antibody, to alleviate symptoms of a CEACAM1-associated disease or disorder. As used herein, "alleviating symptoms of a CEACAM1-associated disease or disorder" refers to improving any condition or symptom associated with a CEACAM1-associated disease or disorder. When compared to an equivalent untreated control, such reduction is at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95%, 99% or more reduction, as measured by any standard method. Various means for administering the compositions described herein to a subject are known to those skilled in the art. Such methods may include, but are not limited to, oral, parenteral, intravenous, intramuscular, subcutaneous, transdermal, airway (aerosol), pulmonary, dermal, topical, injection, or intratumoral administration. Administration may be local or systemic.

[0210] The term "effective amount" as used herein refers to the amount of hCEACAM1 antibody or antigen-binding fragment necessary to alleviate at least one or more symptoms of a disease or disorder, and relates to the amount of a pharmacological composition sufficient to provide a desired effect. Thus, the term "therapeutically effective amount" refers to the amount of a CEACAM1-related disease or disorder sufficient to provide a specific anti-CEACAM1 effect when administered to a typical subject. Effective amount, as used herein, may also include, in various contexts, an amount sufficient to slow down the onset of disease symptoms, an amount sufficient to change the course of a disease symptom (e.g., including but not limited to, delaying the progression of disease symptoms), or an amount sufficient to reverse disease symptoms. Thus, it is generally not possible to specify an exact "effective amount". Meanwhile, for any given case, the appropriate "effective amount" can be determined by those skilled in the art with only routine experimentation.

[0211] Effective doses, toxicity, and therapeutic efficacy can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, to determine the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). Dosages can vary depending on the dosage form employed and the route of administration utilized. The dose ratio between toxic and therapeutic effects is the therapeutic index, which can be expressed as the LD50 / ED50 ratio. Compositions and methods that exhibit large therapeutic indices are preferred. Therapeutically effective doses can be estimated initially from cell culture assays. Doses can also be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e., the concentration of CEACAM1 antibody or antigen-binding fragment that achieves half-maximal inhibition of symptoms) as determined in cell culture or a suitable animal model. Plasma levels can be measured, for example, by high performance liquid chromatography. The effect of any particular dosage can be monitored by a suitable bioassay, such as, for example, an assay for CEACAM1 binding, among others. Dosages can be determined by a physician and adjusted, if necessary, to meet the observed effects of treatment.

[0212] Effective dose, toxicity, and therapeutic efficacy can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, to determine the minimum effective dose and / or maximum tolerated dose. Dosage can vary depending on the dosage form used and the route of administration utilized. The therapeutically effective dose can be initially estimated from cell culture assays. The dose can also be formulated in animal models to achieve a dosage range between the minimum effective dose and the maximum tolerated dose. The effect of any particular dosage can be monitored by suitable bioassays, such as, for example, tumor growth and / or size assays, among others. Dosage can be determined by a physician and adjusted as necessary to meet the observed effect of treatment.

[0213] In some embodiments, the technology described herein relates to a pharmaceutical composition comprising the CEACAM1 antibody or antigen-binding fragment described herein and optionally a pharmaceutically acceptable carrier.In some embodiments, the active ingredient of the pharmaceutical composition comprises the CEACAM1 antibody or antigen-binding fragment described herein.In some embodiments, the active ingredient of the pharmaceutical composition essentially consists of the CEACAM1 antibody or antigen-binding fragment described herein.In some embodiments, the active ingredient of the pharmaceutical composition consists of the CEACAM1 antibody or antigen-binding fragment described herein.Pharmaceutically acceptable carriers and diluents include saline, aqueous buffer, solvent, and / or dispersion medium.The use of such carriers and diluents is well known in the art. Some non-limiting examples of materials that may serve as pharma- ceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, methylcellulose, ethylcellulose, microcrystalline cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) lubricants, such as magnesium stearate, sodium lauryl sulfate, and talc; (8) pharmaceutical additives, such as cocoa butter and suppository waxes. pharmaceutical additives;(9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil;(10) glycols, such as propylene glycol;(11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol (PEG);(12) esters, such as ethyl oleate and ethyl laurate;(13) agar;(14) buffers, such as magnesium hydroxide and aluminum hydroxide;(15) alginic acid;(16) pyrogen-free water;(17) isotonic saline;(18) Ringer's solution;(19) ethyl alcohol;(20) pH buffers;(21) polyesters, polycarbonates, and / or polyanhydrides;(22) bulking agents, such as polypeptides and amino acids;(23) serum components, such as serum albumin, HDL, and LDL;(22) C2-C3 fatty acids, such as ethanol. 12Alcohol; and (23) other non-toxic compatible substances used in pharmaceutical formulations. Wetting agents, coloring agents, releasing agents, coating agents, sweeteners, flavoring agents, perfumes, preservatives, and antioxidants may also be present in the formulation. Terms such as "pharmaceutical additives", "carriers", "pharmaceutical acceptable carriers" or the like are used interchangeably herein. In some embodiments, the carrier inhibits the degradation of the active agent, for example, the CEACAM1 antibody or antigen-binding fragment described herein.

[0214] Therapeutic compositions comprising antibodies or antigen-binding fragments thereof can be administered in any convenient manner, including by injection, infusion, placement, or implantation. The compositions described herein can be administered to a patient subcutaneously, intradermally, intratumorally, intranodally, intramedullary, intramuscularly, intracranially, intravenously or intralymphatically, or intraperitoneally.

[0215] In certain embodiments, the antibody or antigen-binding fragment thereof is administered to the mammal by intravenous infusion, i.e., by introduction of the antibody or antigen-binding fragment thereof into the vein of the mammal for a certain period of time, which in certain embodiments is about 5 minutes, about 10 minutes, about 30 minutes, about 1 hour, about 2 hours, about 4 hours, or about 8 hours.

[0216] In certain embodiments, a dose of the compound or composition is administered to the subject, for example, daily, every other day, every 2-3 days, every other day, once a week, twice a week, three times a week, once every two weeks, or once a month. In other embodiments, two, three, or four doses of the compound or composition are administered to the subject daily, every 2-3 days, every other day, once a week, once every two weeks, or once a month. In some embodiments, a dose(s) of the compound or composition is administered for 2 days, 3 days, 5 days, 7 days, 14 days, 21 days, or 28 days. In certain embodiments, a dose of the compound or composition is administered for 1 month, 1.5 months, 2 months, 2.5 months, 3 months, 4 months, 5 months, 6 months, or longer. The duration of treatment may depend on the outcome or benefit of the treatment.

[0217] Combination therapy In one aspect, the method described herein includes the administration of CEACAM1 antibody agent administered together with additional therapeutic agents. Such additional agents include, but are not limited to, cytotoxic agents, chemotherapeutic agents, growth inhibitors, anti-inflammatory agents, anti-cancer agents, anti-neurodegenerative agents, and anti-infective agents. The agents used in such combination therapy can be divided into one or more of the above-mentioned categories. The administration of the antibody agent and the additional therapeutic agent can be simultaneous or sequential. The administration of the antibody or its antigen-binding fragment and the additional therapeutic agent can be separate or as a mixture. Furthermore, the method of treatment contemplated by the present invention may relate to the treatment in combination with one or more cancer therapies selected from the group of antibody therapy, chemotherapy, cytokine therapy, dendritic cell therapy, gene therapy, hormone therapy, laser light therapy, and radiation therapy, among others.

[0218] Exemplary additional therapeutic agents also include radionuclides with high-energy ionizing radiation that are capable of producing multiple strand breaks in nuclear DNA and are therefore suitable for inducing cell death (e.g., in cancer). Exemplary high-energy radionuclides include: 90 Y, 125 I, 131 I,123 I, 111 In, 105 Rh, 153 Sm, 67 Cu, 67 Ga, 166 Ho, 177 Lu, 186 Re, and 188 These isotopes include Re. These isotopes typically generate short-pathway, high-energy alpha or beta particles. Such radionuclides kill cells in their immediate vicinity, for example, neoplastic cells to which the radionuclides are attached or contained. They have little or no effect on non-localized cells, and are essentially non-immunogenic.

[0219] Exemplary additional therapeutic agents also include cytotoxic agents such as cytostatic agents (e.g., alkylating agents, DNA synthesis inhibitors, DNA intercalators or crosslinkers, or DNA-RNA transcription regulators), enzyme inhibitors, gene regulators, cytotoxic nucleosides, tubulin binding agents, hormones and hormone antagonists, anti-angiogenic agents, and the like.

[0220] Exemplary additional therapeutic agents also include alkylating agents such as the anthracycline drug family (e.g., adriamycin, carminomycin, cyclosporine-A, chloroquine, methopterin, mithramycin, porfiromycin, streptonigrin, anthracenedione, and aziridine). In another embodiment, the chemotherapy moiety is a cytostatic agent such as a DNA synthesis inhibitor. Examples of DNA synthesis inhibitors include, but are not limited to, methotrexate and dichloromethotrexate, 3-amino-1,2,4-benzotriazine 1,4-dioxide, aminopterin, cytosine O-D-arabinofuranoside, 5-fluoro-5'-deoxyuridine, 5-fluorouracil, ganciclovir, hydroxyurea, actinomycin-D, and mitomycin C. Exemplary DNA intercalators or crosslinkers include, but are not limited to, bleomycin, carboplatin, carmustine, chlorambucil, cyclophosphamide, cis-diammineplatinum(II) dichloride (cisplatin), melphalan, mitoxantrone, and oxaliplatin.

[0221] Exemplary additional therapeutic agents also include transcriptional regulators such as cutinomycin D, daunorubicin, doxorubicin, homoharringtonine, and idarubicin. Other exemplary cytostatic agents compatible with the methods described herein include ansamycin benzoquinones, quinonoid derivatives (e.g., quinolones, genistein, bactacyclines), busulfan, ifosfamide, mechlorethamine, triazicon, diazicon, carbazylquinone, indoloquinone EO9, diaziridinyl-benzoquinone methyl DZQ, triethylene phosphoramide, and nitrosourea compounds (e.g., carmustine, lomustine, semustine).

[0222] Exemplary additional therapeutic agents include, for example, cytotoxic nucleosides such as adenosine arabinoside, cytarabine, cytosine arabinoside, 5-fluorouracil, fludarabine, floxuridine, ftorafur, and 6-mercaptopurine; taxoids (e.g., paclitaxel, docetaxel, taxanes), nocodazole, rhizoxin, dolastatins (e.g., dolastatin-10, dolastatin-11, or dolastatin-15), colchicine, and colchicinoids. (e.g., ZD6126), combretastatins (e.g., combretastatin A-4, AVE-6032), and tubulin binding agents such as vinca alkaloids (e.g., vinblastine, vincristine, vindesine, and vinorelbine); and antiangiogenic compounds such as angiostatin K1-3, DL-a-difluoromethyl-ornithine, endostatin, fumagillin, genistein, minocycline, staurosporine, and (±)-thalidomide.

[0223] Exemplary additional therapeutic agents also include hormones and hormone antagonists such as corticosteroids (e.g., prednisone), progestins (e.g., hydroxyprogesterone or medroprogesterone), estrogens, (e.g., diethylstilbestrol), antiestrogens (e.g., tamoxifen), androgens (e.g., testosterone), aromatase inhibitors (e.g., aminoglutethimide), 17-(allylamino)-17-demethoxygeldanamycin, 4-amino-1,8-naphthalimide, apigenin, brefeldin A, cimetidine, dichloromethylene-diphosphonic acid, leuprolide (leuprorelin), luteinizing hormone-releasing hormone, pifithrin-a, rapamycin, sex hormone-binding globulin, and thapsigargin.

[0224] Exemplary additional therapeutic agents also include enzyme inhibitors such as S(+)-camptothecin, curcumin, (-)-deguelin, 5,6-dichlorobenzo-imidazole ribofuranoside, etoposide, formestane, fostriecin, hispidin, 2-imino-l-imidazolidine acetic acid (cyclocreatine), mevinolin, trichostatin A, tyrophostin AG34, and tyrophostin AG879. Exemplary additional therapeutic agents also include enzyme inhibitors such as 5-aza-2 1 -Gene regulators such as deoxycytidine, 5-azacytidine, cholecalciferol (vitamin D3), 4-hydroxytamoxifen, melatonin, mifepristone, raloxifene, trans-retinal (vitamin A aldehyde), retinoic acid, vitamin A acid, 9-cis-retinoic acid, 13-cis-retinoic acid, retinol (vitamin A), tamoxifen, and troglitazone.

[0225] Exemplary additional therapeutic agents also include cytotoxic agents such as, for example, the pteridine drug family, diynenes, and podophyllotoxins. Particularly useful members of such classes include, for example, methopterin, podophyllotoxin, or podophyllotoxin derivatives such as etoposide or etoposide phosphate, leurocidin, vindesine, leurosine, and the like.

[0226] Still other additional therapeutic agents compatible with the teachings herein include auristatins (e.g., auristatin E and monomethylauristan E), calicheamicin, gramicidin D, maytansanoids (e.g., maytansine), neocarzinostatin, topotecan, taxanes, cytochalasin B, ethidium bromide, emetine, tenoposide, colchicine, dihydroxyanthracin dione, mitoxantrone, procaine, tetracaine, lidocaine, propranolol, puromycin, and analogs or homologs thereof.

[0227] In one embodiment, the CEACAM antibody agent is administered in combination with an agent that is a checkpoint inhibitor. Such inhibitors may include small molecule inhibitors, or may include antibodies or antigen-binding fragments that bind to and block or inhibit immune checkpoint receptors, or antibodies that bind to and block or inhibit immune checkpoint receptor ligands. Exemplary checkpoint molecules that can be targeted for blockade or inhibition include CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, GAL9, LAG3, TIM-3, VISTA, KIR, 2B4 (which belongs to the CD2 molecule family and is involved in the activation of all NK cells, y6 cells, and memory CD8 cells). + (4) T cells), CD160 (also called BY55), CGEN-15049, CHK1 and CHK2 kinases, A2aR, and various B-7 family ligands. B7 family ligands include, but are not limited to, B7-1, B7-2, B7-DC, B7-H1, B7-H2, B7-H3, B7-H4, B7-H5, B7-H6, and B7-H7. Checkpoint inhibitors include antibodies or antigen-binding fragments thereof, other binding proteins, biotherapeutics, or small molecules that bind to and block or inhibit the activity of one or more of CTLA-4, PDL1, PDL2, PD1, BTLA, HVEM, TIM-3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, and CGEN-15049. Illustrative immune checkpoint inhibitors include tremelimumab (a CTLA-4 blocking antibody), anti-OX40, and Yervoy / ipilimumab (anti-CTLA-4 checkpoint inhibitors), as well as the PD-1 and PD-L1 inhibitors mentioned above. Checkpoint protein ligands include, but are not limited to, PD-L1, PD-L2, B7-H3, B7-H4, CD28, and CD86.

[0228] In some embodiments, the CEACAM1 antibody agent is administered in combination with an agent that inhibits TIGIT, LAP, podoplanin, protein C receptor, ICOS, GITR, CD226, or CD160.

[0229] In some embodiments, the CEACAM1 antibody agent is administered in combination with an agent that inhibits CTLA-4, PD-1, PD-L1, or PD-L2. In some embodiments, the CEACAM1 antibody agent is administered in combination with an agent that inhibits TIM-3.

[0230] It will be understood that the invention is not limited to the specific molecules, compositions, methodologies, or protocols described, as they may vary. In practicing or testing the embodiments of the invention, any methods and materials similar or equivalent to those described herein can be used. It will be further understood that the disclosure of the invention herein includes all possible combinations of such specific features. For example, if a specific feature is disclosed in the context of a particular aspect or embodiment of the invention, or in a particular claim, that feature can be used in combination and / or in the context of other specific aspects and embodiments of the invention, and also in the invention generally, to the extent possible.

[0231] When reference is made herein to a method including two or more specified steps, the specified steps may be performed in any order or simultaneously (unless the context excludes this possibility), and the method may include one or more other steps that are performed before any specified step, between two of the specified steps, or after all the specified steps (unless the context excludes this possibility). EXAMPLES

[0232] Example 1: Companion diagnostics for therapeutic agents directed against human CEACAM1 A therapeutic monoclonal antibody candidate has been developed for human treatment (CP08; see, e.g., PCT / US2019 / 065212 CP08H03 / Vk8 S29A or CP08H03 / CP08F05). The structural basis for this antibody's binding to its target protein, human carcinoembryonic antigen cell adhesion molecule 1 (hCEACAM1), has been analyzed. These studies have shown that amino acids such as glutamine 89 (Q89) and tyrosine 34 (Y34) are involved in this antibody (CP08) binding. These two amino acids are affected by allelic variation, including the conversion of glutamine 89 to histidine (H) and tyrosine 34 to cysteine ​​(C). In addition, the alanine 49 residue is located near the CP08 contact site and was found to be affected by allelic variation (alanine 49 to valine (V)). CEACAM1 is unique in relation to other similar hCEACAM members (hCEACAM3, hCEACAM5, hCEACAM6) as it contains a valine at position 49. It was predicted that the valine at position 49 would affect CP08 binding. In addition to these SNPs (A49V, Q89H, Y34C), glutamine 44 to leucine (Q44L) is also contained within a continuous patch in the center of the CP08 antibody binding surface of hCEACAM1. Q44L may also affect drug binding to hCEACAM1 based on studies with the mouse parent of hCEACAM1 (5F4). This suggests that humans expressing natural variants at these positions may bind CP08 antibodies poorly and therefore respond poorly (or refractory) to treatment with this therapeutic antibody. This hypothesis was tested by generating an A49V and Q89H double mutant of hCEACAM1.

[0233] X-ray crystallography revealed that the A49V mutation causes a rotamer shift of the phenylalanine (F) 29 residue, which predicts the formation of a steric clash. This double mutant (A49V, Q89H) was found to be unable to bind CP08. This predicts that humans expressing one or both of these mutations would respond poorly or be refractory to CP08-based therapeutics, respectively. Furthermore, we modeled these four allelic variants on the surface of hCEACAM1 using a previously published hCEACAM1 structure. We found that all four of these allelic variants reside within a continuous patch located in the center of the GFCC' face where CP08 binds. Mutation of hCEACAM1 to the A48V / Q89H double mutant was found to abrogate CP08 binding to hCEACAM1. Recent studies suggest that 14% of Caucasians and 43% of Africans express the A49V and Q89H haplotypes, and 7% and 21% of these populations, respectively, are expected to be unable to respond to CP08-based drugs. Moreover, Q44L and Y34C also affect the binding of CP08 to hCEACAM1. Furthermore, it is expected that all therapeutic antibodies directed against hCEACAM1 that bind to the center of the GFCC' face will be affected by these mutations.

[0234] There is no guidance on how to accurately apply therapeutic antibodies against hCEACAM1 to populations in clinical use to predict responsive / non-responsive populations. It has not been previously recognized that human allelic variants that characterize the hCEACAM1 structure can be used as a means to determine responsiveness to anti-hCEACAM1 therapies and drugs that target the GFCC' face of human CEACAM1, the major ligand binding site. If an individual carries a particular allelic variant, binding of anti-hCEACAM1 therapeutic antibodies to its target (hCEACAM1) is expected to be absent (homozygous A49V / Q89H haplotype) or reduced (heterozygous A49V, heterozygous Q89H, or heterozygous or heterozygous Y34C and Q44L carriers). Knowing that a patient does or does not carry one of these variants allows for the selection of the treatment most likely to work for a given subject. If the subject is found to be homozygous wild type, then the CEACAM1-associated disease or disorder is likely to respond to treatment with a CP08-based antibody.

[0235] Group 1 patients are subjects who are found to be compound homozygous for the A49V and Q89H variants. These subjects are considered hCEACAM1 therapeutic "non-binders" and therefore unlikely to respond to therapeutic agents; these individuals would not be considered suitable for therapeutic administration of CP08-based therapy and would also likely be non-responders with any antibody that specifically binds to the GF-CC' face of wild-type hCEACAM1.

[0236] Group 2 patients are subjects who are found to be heterozygous for A49V and / or Q89H, or carriers of either zygosity Y34C or Q44L. In this case, such subjects are considered to be likely therapeutic candidates for administration of CP08-based antibody treatment, but may require increased dosages of anti-hCEACAM1 antibody treatment to demonstrate a response. In one embodiment, the dose of CP08-based antibody agent for such patients may be adjusted based on an assessment of the affinity of binding of the CP08-based antibody agent to the subject's hCEACAM1 (see below).

[0237] Group 3 patients are human subjects who have a homozygous wild-type configuration of the hCEACAM1 gene and thus the protein. These latter subjects are considered to be therapeutic candidates who are likely to benefit from treatment with a CP08-based antibody agent.

[0238] Groups 1 and 2 can be subjected to a second level of characterization, where tissue samples (blood and / or tumor) are evaluated for binding to the drug. Subjects in groups 1 and 2 who do not show evidence of binding to the therapeutic agent in such tissue sample testing (possibly due to the absence of hCEACAM1 protein, or a proportion of allelic variants that cannot bind, e.g., low levels of hCEACAM1 expression combined with specific genotypic variants) are not considered candidates for treatment. Thus, such findings are believed to inform treatment decision-making.

[0239] To define the genotype of such allelic variants, a body fluid (e.g., blood, saliva) or tissue sample (e.g., oral swab and / or tumor tissue) is obtained. DNA is extracted from the body fluid or tissue sample. A tissue sample (blood or tumor tissue) is prepared to define therapeutic antibody binding, which is known to those skilled in the art.

[0240] To define the genotype of a subject, DNA from the subject is evaluated for the presence of A49V, Q89H, Q44L and Y34C alleles, and their zygosity (homozygous or heterozygous) is determined. The methods used to achieve this may include Taqman assay, amplicon sequencing (all four single nucleotide polymorphisms are present on a single exon that codes for a protein of 110 amino acids, and therefore is approximately 300 bp), oligonucleotide binding determination, or other assays. Such methods are well known in the art and do not need to be detailed here. Individuals who are found to be compound homozygous for A49V and Q89H variants (group 1) are considered hCEACAM1 therapeutic non-binders, and therefore are likely to be non-responsive to treatment; these individuals are deemed unsuitable for administration of CP08-based antibody drug treatment, and will not be further tested. Such individuals would not be treated with an agent that is unlikely to provide benefit and would therefore benefit from early treatment with another anti-cancer agent, whether that be a different anti-CEACAM1 agent, one or more chemotherapy, immunotherapy, radiation, surgery, or any combination thereof.

[0241] Human subjects that are determined to be in group 2 or group 3 based on their genotype can then be evaluated for binding of drugs to their tissues or CEACAM protein extracted from their tissues. In this case, for example, cells from peripheral blood are evaluated in solution or after being mounted on a fixed surface (e.g., glass slide, matrix) for solution-based assay (flow cytometry, mass cytometry) or surface-based assay (immunohistochemistry [IHC] or immunofluorescence [IF]) to detect drug binding compared to a positive control that detects hCEACAM1 protein regardless of genotype. These techniques are also well known in the art and do not need to be detailed here. There are many antibodies that can be used for genotype-independent binding, including, for example, commercially available antibodies that recognize hCEACAM1 domains other than IgV domain. Tumor tissue can either be mechanically separated for solution-based assay as described above, or mounted in a matrix (e.g., OCT, formalin / paraffin) that allows therapeutic antibody binding to be compared to control by IHC or IF. In such assays, quantification can be performed to determine the cutoff of binding of the therapeutic antibody compared to the control antibody using tissues from genotypically characterized samples expressing hCEACAM1, cell lines transfected with genotypically defined proteins expressing wild type and allelic variants (e.g., human embryonic kidney 293T cells, HeLa cells), or cell lines expressing wild type hCEACAM1 and / or genotypic variants generated by Crispr-Cas9-mediated mutation or other methods as controls. Such transfected or engineered cell lines can be characterized for their binding to the therapeutic antibody (compared to the control antibody) by solution or matrix-based techniques using specific amounts of the therapeutic or control antibody to establish the expected lower limit of detection.

[0242] In an alternative approach, proteins can be extracted from tissues and assessed for binding to therapeutic antibodies compared to controls using ELISA, immunoblot, surface plasmon resonance, biolayer interference, or similar types of immunobinding assays that compare the signal detected with fixed concentrations of wild-type or mutant proteins (including combinations of various genotypes, in the latter case).

[0243] Subjects in groups 2 and 3 who have no binding to the drug and / or control (due to lack of hCEACAM1 expression or hCEACAM1 expression levels associated with a particular genotype showing reduced binding), or who show a particular level of binding to the drug below a certain limit, are not considered therapeutic candidates for treatment.

[0244] Thus, a method is disclosed for determining whether a patient is responsive or non-responsive to anti-hCEACAM1 therapeutic antibody administration. In this method, an individual who is considered for treatment with anti-human CEACAM1 therapeutic agent is examined for the expression of wild-type or mutant (allelic variant) of hCEACAM1 (A49V, Q89H, Q44L, and Y34C allele). Individuals who are compound homozygous for A49V and Q89H variants are considered hCEACAM1 therapeutic "non-binders" and are unlikely to respond to therapeutic agents. Individuals who are found to be heterozygous for A49V and / or Q89H variants, or carriers of Y34C or Q44L of either zygosity, may be binders and may respond to therapeutic agents when administered at appropriate dosages. Individuals with an hCEACAM1 gene in the wild-type configuration, i.e., the gene lacks the A49V, Q89H, Q44L, and Y34C variant alleles and therefore the hCEACAM1 protein is wild-type, are likely to respond to anti-hCEACAM1 monoclonal antibody therapy.

[0245] Anti-CEACAM1 antibodies and anti-CEACAM1 monoclonal antibodies are described in US Patent 9,556,271, WO2013 / 082366, and WO2020 / 118295, the latter of which describes CP08. For example, additional anti-CEACAM1 antibodies that may be used as alternatives for subjects for whom CP08-based Abs are ineffective are described in US2016 / 0376374 and US2020 / 0277398. Although the present invention is disclosed in certain embodiments, it will be understood by those skilled in the art that certain substitutions, modifications, and / or omissions may be made to the embodiments without departing from the spirit of the invention. Thus, the foregoing description is intended to be illustrative only and should not limit the scope of the present invention. All references, scientific papers, patent publications, and any other documents cited herein are incorporated herein by reference for the material disclosed therein.

Claims

1. (i) receiving the results of a hCEACAM1 genotyping assay performed on a sample from a subject in need of treatment for a CEACAM1-associated disease or disorder; and (ii) if the subject's hCEACAM1 genotype is homozygous wild type for A49, Q89, Q44, and Y34, or heterozygous for A49V, Q89H, Q44L, and / or Y34C, selecting treatment with an agent comprising an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1; and If the subject's hCEACAM1 genotype is homozygous for the A49V, Q89H, Q44L, and / or Y34C hCEACAM1 alleles, selecting treatment with an agent other than an antibody that binds to an epitope in the GF-CC' domain of CEACAM1 or an agent that contains an antigen-binding domain. The method includes:

2. (i) genotyping a sample from a subject in need of treatment for a CEACAM1-associated disease or disorder to detect the subject's hCEACAM1 genotype; and (ii) if the subject's hCEACAM1 genotype is homozygous wild type for A49, Q89, Q44, and Y34, or heterozygous for A49V, Q89H, Q44L, and / or Y34C, selecting treatment with an agent comprising an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1; and If the subject's hCEACAM1 genotype is homozygous for the A49V, Q89H, Q44L, and / or Y34C hCEACAM1 alleles, selecting treatment with an agent other than an antibody that binds to an epitope in the GF-CC' domain of CEACAM1 or an agent that contains an antigen-binding domain. The method includes:

3. 3. The method of claim 1 or claim 2, wherein the subject has or has been diagnosed with a chronic disease.

4. 5. The method of claim 4, wherein the chronic disease is selected from the group consisting of cancer; persistent infection; and chronic viral infection.

5. The method of any one of claims 1 to 5, wherein the subject is a human.

6. an antibody or antigen-binding domain that binds to an epitope of the GF-CC' domain of CEACAM1, comprising a heavy chain variable region and a light chain variable region; the heavy chain variable region and the light chain variable region each comprise CDR1, CDR2, and CDR3; The sequence of CDR1H is X 1 HX 2 X 3 S (SEQ ID NO:1); X 1 is A, D, N, or S; X 2 is A or G; X 3 is an amino acid having a hydrophobic side chain containing I or M; The sequence of CDR2H is The sequence: The sequence of CDR3H is The sequence: X 4 is D, G, or P; X 5 is F or P; X 6 is D or F; X 7 is A or Y; X 8 is L, H, or F; The sequence of CDR1L comprises the sequence RANSAVSYMY (SEQ ID NO: 4); The sequence of CDR2L comprises the sequence LTSNRAT (SEQ ID NO: 5); The sequence of CDR3L is The sequence: X 9 is W or N; X 10 is S or T; X 11 is A or an amino acid having a neutral hydrophilic side chain, including S, N, and T; X 12 is L, F, or N; X 13 is P or F, The method according to any one of claims 1 to 5.

7. The method of any one of claims 1 to 5, wherein the sample is selected from a blood sample, a tissue sample, a tumor sample, and a cell sample.

8. 6. The method of any one of claims 1 to 5, wherein the genotyping assay comprises direct probing, allele-specific hybridization, PCR, allele-specific amplification (ASA), single base extension, ARMS-PCR, Taqman, oligo ligation assay, single strand conformation analysis (SSCP), Genetic Bit Analysis and RFLP direct sequencing, mass spectrometry (MALDI-TOF), and DNA array.

9. The method of any one of claims 1 to 5, further comprising administering the selected treatment.

10. The method of any one of claims 1 to 9, further comprising the steps of: assaying a sample from the subject containing CEACAM1 to determine the extent to which the subject's CEACAM1 is bound by an agent comprising an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1 if the CEACAM1 genotype is heterozygous for A49V, Q89H, Q44L, and / or Y34C; and adjusting the amount and / or dosage of the agent to be administered as a function of the degree of binding of the agent to the subject's CEACAM1.

11. 11. The method of claim 10, wherein said assaying step comprises a method selected from flow cytometry, mass cytometry, immunohistochemistry, immunofluorescence or FRET assay, surface plasmon resonance, radioimmunoassay, and biolayer interferometry.

12. The method of claim 2, further comprising the step of extracting genomic DNA from a sample obtained from the subject prior to the genotyping step.

13. 1. A method of treating a subject in need of treatment for a CEACAM1 associated disease or disorder, comprising: (i) genotyping a sample from the subject to detect the subject's hCEACAM1 genotype; and (ii) administering a therapeutically effective amount of an agent comprising an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1 if the subject's hCEACAM1 genotype is homozygous wild type for A49, Q89, Q44, and Y34; and (iii) if the subject's hCEACAM1 genotype is heterozygous for A49V, Q89H, Q44L, and / or Y34C, assaying a sample from the subject containing CEACAM1 to determine the extent to which the subject's CEACAM1 is bound by an agent comprising an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1, and administering a therapeutically effective amount of the agent, wherein the amount or dosing schedule of the agent administered is adjusted as a function of the extent of binding of the agent to the subject's CEACAM1; and (iv) administering a therapeutically effective amount of treatment with an agent other than an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1 if the subject's hCEACAM1 genotype is homozygous for A49V, Q89H, Q44L, and / or Y34C. thereby treating the CEACAM1-associated disease or disorder.

14. 14. The method of claim 13, wherein the subject has or has been diagnosed with a chronic disease.

15. 15. The method of claim 14, wherein the chronic disease is selected from the group consisting of cancer; persistent infection; and chronic viral infection.

16. The method of any one of claims 13 to 15, wherein the subject is a human.

17. an antibody or antigen-binding domain that binds to an epitope of the GF-CC' domain of CEACAM1, comprising a heavy chain variable region and a light chain variable region; the heavy chain variable region and the light chain variable region each comprise CDR1, CDR2, and CDR3; The sequence of CDR1H is X 1 HX 2 X 3 S (SEQ ID NO:1); X 1 is A, D, N, or S; X 2 is A or G; X 3 is an amino acid having a hydrophobic side chain containing I or M; The sequence of CDR2H is The sequence: The sequence of CDR3H is The sequence: X 4 is D, G, or P; X 5 is F or P; X 6 is D or F; X 7 is A or Y; X 8 is L, H, or F; The sequence of CDR1L comprises the sequence RANSAVSYMY (SEQ ID NO: 4); The sequence of CDR2L comprises the sequence LTSNRAT (SEQ ID NO: 5); The sequence of CDR3L is The sequence: X 9 is W or N; X 10 is S or T; X 11 is A or an amino acid having a neutral hydrophilic side chain, including S, N, and T; X 12 is L, F, or N; X 13 is P or F, The method of any one of claims 13 to 16.

18. The method of any one of claims 13 to 17, wherein the sample is selected from a blood sample, a tissue sample, a tumor sample, and a cell sample.

19. The method of any one of claims 13 to 18, wherein the genotyping assay comprises direct probing, allele-specific hybridization, PCR, allele-specific amplification (ASA), single base extension, ARMS-PCR, Taqman, oligoligation assay, single-strand conformation analysis (SSCP), gene bit analysis and RFLP direct sequencing, mass spectrometry (MALDI-TOF), and DNA array.

20. The method of any one of claims 13 to 19, wherein the step of assaying a sample from the subject containing CEACAM1 to determine the extent to which the subject's CEACAM1 is bound by the agent comprises a method selected from flow cytometry, mass cytometry, immunohistochemistry, immunofluorescence or FRET assay, surface plasmon resonance, radioimmunoassay, and biolayer interferometry.

21. (i) extracting genomic DNA from a biological sample from a subject in need of treatment for a CEACAM1-associated disease or disorder; (ii) genotyping the subject's DNA for sequences corresponding to CEACAM A49, Q89, Q44, and Y34, thereby determining whether the subject is homozygous wild type for A49, Q89, Q44, and Y34, or homozygous or heterozygous for each of A49V, Q89H, Q44L, and Y34C. The assay comprising:

22. 22. The assay of claim 21, wherein the CEACAM1-associated disease or disorder is selected from the group consisting of cancer; persistent infection; and chronic viral infection.

23. 23. The assay of claim 21 or 22, wherein the subject is a human.

24. The assay of any one of claims 21 to 23, wherein the biological sample is selected from a blood sample, a tissue sample, a tumor sample, and a cell sample.

25. The assay of any one of claims 21 to 24, wherein the genotyping comprises direct probing, allele-specific hybridization, PCR, allele-specific amplification (ASA), single base extension, ARMS-PCR, Taqman, oligoligation assay, single-stranded conformation analysis (SSCP), gene bit analysis and RFLP direct sequencing, mass spectrometry (MALDI-TOF), and DNA arrays.

26. (iii) assaying the subject CEACAM1 protein to determine the extent to which the subject CEACAM1 is bound by an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1.

26. The assay of any one of claims 21 to 25, further comprising:

27. 28. The assay of claim 27, wherein the assaying step (iii) comprises a method selected from flow cytometry, mass cytometry, immunohistochemistry, immunofluorescence or FRET assay, surface plasmon resonance, radioimmunoassay, and biolayer interferometry.

28. (i) extracting a protein comprising CEACAM1 from a biological sample from a subject in need of treatment for a CEACAM1-associated disease or disorder; and (ii) assaying the subject CEACAM1 in the protein extract to determine the extent to which the subject CEACAM1 is bound by an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1. The assay comprising:

29. 30. The assay of claim 28, wherein the assaying step (ii) comprises a method selected from flow cytometry, mass cytometry, immunohistochemistry, immunofluorescence or FRET assay, surface plasmon resonance, radioimmunoassay, and biolayer interferometry.

30. (iii) genotyping the subject's DNA for sequences corresponding to CEACAM A49, Q89, Q44, and Y34, thereby determining whether the subject is homozygous wild type for A49, Q89, Q44, and Y34, or homozygous or heterozygous for each of A49V, Q89H, Q44L, and Y34C.

30. The assay of claim 28 or 29, further comprising:

31. 31. The assay of claim 30, wherein the genotyping step comprises direct probing, allele-specific hybridization, PCR, allele-specific amplification (ASA), single base extension assay, ARMS-PCR, Taqman, oligoligation assay, single-stranded conformation analysis (SSCP), gene bit analysis and RFLP direct sequencing, mass spectrometry (MALDI-TOF), and DNA arrays.

32. 1. A method of selecting said hCEACAM1 antibody therapy for a subject in need thereof, comprising: analyzing genomic DNA from a biological sample obtained from the subject for the subject's genotype for alleles of hCEACAM1 at sites corresponding to A49, Q89, Q44, and Y34; and If the genotype is found to be homozygous for A49, Q89, Q44, and Y34, or heterozygous for one or more of the A49V, Q89H, Q44L, and Y34C alleles of CEACAM1, selecting treatment with an agent comprising an hCEACAM1 antibody or an antigen-binding fragment thereof that binds to an epitope in the GF-CC' domain of CEACAM1; or If the genotype is found to be homozygous for one or more of the A49V, Q89H, Q44L, and Y34C alleles of CEACAM1, selecting a treatment with an agent other than an agent comprising an hCEACAM1 antibody or an antigen-binding fragment thereof that binds to an epitope in the GF-CC' domain of CEACAM1. The method comprising:

33. 33. The method of claim 32, wherein the subject has or has been diagnosed with a chronic disease.

34. 34. The method of claim 33, wherein the chronic disease is selected from the group consisting of cancer; persistent infection; and chronic viral infection.

35. The method of any one of claims 32 to 34, wherein the subject is a human.

36. an antibody or antigen-binding domain that binds to an epitope of the GF-CC' domain of CEACAM1, comprising a heavy chain variable region and a light chain variable region; the heavy chain variable region and the light chain variable region each comprise CDR1, CDR2, and CDR3; The sequence of CDR1H is X 1 HX 2 X 3 S (SEQ ID NO:1); X 1 is A, D, N, or S; X 2 is A or G; X 3 is an amino acid having a hydrophobic side chain containing I or M; The sequence of CDR2H is The sequence: The sequence of CDR3H is The sequence: X 4 is D, G, or P; X 5 is F or P; X 6 is D or F; X 7 is A or Y; X 8 is L, H, or F; The sequence of CDR1L comprises the sequence RANSAVSYMY (SEQ ID NO: 4); The sequence of CDR2L comprises the sequence LTSNRAT (SEQ ID NO: 5); The sequence of CDR3L is The sequence: X 9 is W or N; X 10 is S or T; X 11 is A or an amino acid having a neutral hydrophilic side chain, including S, N, and T; X 12 is L, F, or N; X 13 is P or F, The method of any one of claims 32 to 35.

37. The method of any one of claims 32 to 36, wherein the sample is selected from a blood sample, a tissue sample, a tumor sample, and a cell sample.

38. The method of any one of claims 32 to 37, wherein the genotype analysis comprises direct probing, allele-specific hybridization, PCR, allele-specific amplification (ASA), single base extension, ARMS-PCR, Taqman, oligoligation assay, single-strand conformation analysis (SSCP), gene bit analysis and RFLP direct sequencing, mass spectrometry (MALDI-TOF), and DNA array.

39. The method of any one of claims 32 to 38, further comprising administering the selected treatment.

40. The method of any one of claims 32 to 39, further comprising the steps of: assaying a sample from the subject containing CEACAM1 to determine the extent to which the subject's CEACAM1 is bound by an agent comprising an antibody or antigen-binding domain that binds to an epitope in the GF-CC' domain of CEACAM1 if the CEACAM1 genotype is heterozygous for A49V, Q89H, Q44L, and / or Y34C; and adjusting the amount and / or dosage of the agent to be administered as a function of the extent of binding of the agent to the subject's CEACAM1.

41. 41. The method of claim 40, wherein said assaying step comprises a method selected from flow cytometry, mass cytometry, immunohistochemistry, immunofluorescence or FRET assay, surface plasmon resonance, radioimmunoassay, and biolayer interferometry.

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