CEA assay for patient selection in cancer treatment

Abstract

Methods for identifying and treating patients with cancers expressing CEACAM5 are provided. The methods include measuring circulating CEA to identify patients who are likely to benefit from treatment with a CEACAM5-specific agent. Such patients may have high circulating CEA and only low or moderate expression of CEACAM5 on tumor cells as measured by immunohistochemistry (IHC). The CEACAM5-specific agent may be tusamitama ravtansine. Such agents may be used in combination with one or more additional agents to treat cancer. In certain embodiments, the cancer is non-squamous non-small cell lung cancer (NSQ NSCLC).

Description

[Technical field]

[0001] Sequence Listing Reference This application contains a Sequence Listing that has been submitted electronically in xml format, which is incorporated herein by reference in its entirety. The .xml copy created on November 28, 2022 is named PR94291_S287_WO_SANOFI_SEQUENCES.xml.

[0002] The present disclosure relates to the field of cancer treatment. [Background technology]

[0003] Despite recent advances in the treatment of cancer, new effective therapies are still needed when disease progresses after first-line treatment. Current treatment approaches for subsequent systemic options, combining angiogenesis inhibitors with systemic cytotoxic agents such as docetaxel, are accompanied by severe hematological and other toxicities. Docetaxel, pemetrexed or gemcitabine used as single cytotoxic agents provide very limited alternative options. Therefore, targeted cytotoxic therapy may provide improved safety and tolerability as well as efficacy.

[0004] One feature that can be used to target some tumor cells is the surface expression of carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5), which was first described in 1965 as a tumor-associated antigen in human colon cancer tissue extracts (Gold P et al., J Exp Med. 1965; 122(3): 467-481). High levels of CEACAM5 expression have since been observed in several epithelial tumors, but in normal adult tissues, its expression is restricted to a few tissues (Hammarstrom S., Semin Cancer Biol. 1999; 9(2): 67-81; Thompson JA., Tumor Biol. 1995; 16(1): 10-16).

[0005] Tusamitama ravtansine is an antibody against CEACAM5 conjugated to the cytotoxic maytansinoid agent N2'-deacetyl-N-2'(4-methyl-4-mercapto-1-oxopentyl)-maytansine (DM4). Ongoing studies have demonstrated promising preliminary antitumor activity of tusamitama ravtansine in participants heavily pretreated for NSQ NSCLC (TED13751). Summary of the Invention [Means for solving the problem]

[0006] The present disclosure is based, at least in part, on the observation that certain levels of circulating carcinoembryonic antigen (CEA) are useful as a biomarker for selecting patients for treatment of cancers that typically express CEA cell adhesion molecule 5 (CEACAM5) on their cells.

[0007] One aspect of the present disclosure is a method of treating cancer in a subject, comprising: Measuring circulating carcinoembryonic antigen (CEA) in a subject in need of treatment for cancer; administering to the subject an effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody if the subject has circulating CEA of 5 ng / mL or greater (5 μg / L or greater); thereby to treat cancer.

[0008] One aspect of the present disclosure is circulating carcinoembryonic antigen (CEA) for use as a biomarker in the methods disclosed herein.

[0009] In some embodiments, circulating carcinoembryonic antigen (CEA) may be for use as a biomarker in methods of treating cancer in a subject, as disclosed herein.

[0010] In some embodiments, circulating carcinoembryonic antigen (CEA) may be for use as a biomarker in methods of diagnosing cancer in a subject, as disclosed herein.

[0011] One aspect of the disclosure is circulating carcinoembryonic antigen (CEA) for use as a biomarker in a method of treating cancer in a subject, the method comprising: Measuring circulating carcinoembryonic antigen (CEA) in a subject in need of treatment for cancer; administering to the subject an effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody if the subject has circulating CEA of about 5 ng / mL or greater; thereby treating cancer.

[0012] In the uses disclosed herein, the biomarkers are measured in an isolated biological sample.

[0013] In some embodiments, the present disclosure relates to an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent for use in a method of treating cancer in a subject in need thereof, The anti-CEACAM5 antibody comprises an HCDR1 having the amino acid sequence of SEQ ID NO:1, an HCDR2 having the amino acid sequence of SEQ ID NO:2, an HCDR3 having the amino acid sequence of SEQ ID NO:3, an LCDR1 having the amino acid sequence of SEQ ID NO:4, an LCDR2 having the amino acid sequence NTR, and an LCDR3 having the amino acid sequence of SEQ ID NO:5, the cytotoxic agent is a maytansinoid or a maytansinoid analog, and The method includes measuring circulating carcinoembryonic antigen (CEA) in the subject, and if circulating CEA in the subject is greater than or equal to about 5 ng / mL, administering to the subject an effective amount of the ADC, thereby treating cancer.

[0014] In some embodiments, the present disclosure relates to the use of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent for the manufacture of a medicament for use in a method of treating cancer in a subject in need thereof, The anti-CEACAM5 antibody comprises an HCDR1 having the amino acid sequence of SEQ ID NO:1, an HCDR2 having the amino acid sequence of SEQ ID NO:2, an HCDR3 having the amino acid sequence of SEQ ID NO:3, an LCDR1 having the amino acid sequence of SEQ ID NO:4, an LCDR2 having the amino acid sequence NTR, and an LCDR3 having the amino acid sequence of SEQ ID NO:5, the cytotoxic agent is a maytansinoid or a maytansinoid analog, and The method includes measuring circulating carcinoembryonic antigen (CEA) in the subject, and if circulating CEA in the subject is greater than or equal to about 5 ng / mL, administering an effective amount of the pharmaceutical agent to the subject, thereby treating the cancer.

[0015] "Biomarker" is intended to refer to a biological molecule, e.g., a protein or metabolite, that is differentially present, increased or decreased in a biological sample obtained from a subject or group of subjects having a first phenotype, e.g., having a disease, such as cancer, compared to a biological sample from a subject or group of subjects having a second phenotype, e.g., not having the disease. In use, the biomarker is isolated from the subject.

[0016] A "sample" or "biological sample" is intended to refer to biological material isolated from a subject. A biological sample may contain any biological material suitable for detecting a biomarker, i.e., circulating carcinoembryonic antigen (CEA), and may include cells and / or non-cellular material from a subject. A sample may be isolated from any suitable biological tissue or body fluid, such as, for example, kidney tissue, blood, blood plasma (plasma), blood serum (serum), urine, or cerebrospinal fluid (CSF). In some embodiments, the biological sample is a plasma (plasma) or serum (serum) sample.

[0017] In the use disclosed herein, the circulating carcinoembryonic antigen (CEA) measured in a subject who needs to treat cancer can be compared with a reference value. The reference value can be 5ng / mL. A circulating carcinoembryonic antigen (CEA) measured in a subject of about 5ng / mL or more can indicate cancer.

[0018] "Reference value" is intended to refer to a level of circulating CEA that is indicative of a particular disease state, phenotype, such as cancer or lack thereof, as well as combinations of disease states, phenotypes, or lack thereof, in a relevant subject.

[0019] One aspect of the disclosure is a method of diagnosing cancer in a subject, the method comprising at least: a) measuring in a biological sample the amount of circulating carcinoembryonic antigen (CEA) in said subject; b) comparing the measured amount obtained in step a) with a reference value, said reference value being 5 ng / mL; wherein a measurement obtained in step a) of about 5 ng / mL or greater is indicative of cancer.

[0020] In certain embodiments, the anti-CEACAM5 antibody is tusamitamab.

[0021] In certain embodiments, the ADC is tusamitamaravtansine.

[0022] In certain embodiments, the cancer is selected from the group consisting of colorectal cancer, gastric cancer, gastroesophageal junction cancer, esophageal cancer, lung cancer, cervical cancer, pancreatic cancer, ovarian cancer, thyroid cancer, bladder cancer, endometrial cancer, breast cancer, liver cancer, biliary tract cancer (e.g., cholangiocarcinoma), prostate cancer, and skin cancer.

[0023] In certain embodiments, the cancer is gastric cancer, gastroesophageal junction cancer, or esophageal cancer.

[0024] In certain embodiments, the cancer is gastric cancer.

[0025] In certain embodiments, the cancer is lung cancer.

[0026] In certain embodiments, the cancer is non-squamous non-small cell lung cancer (NSQ NSCLC).

[0027] In certain embodiments, the cancer is advanced or metastatic.

[0028] In certain embodiments, the NSQ NSCLC is advanced or metastatic.

[0029] In certain embodiments, circulating CEA is measured after performing immunohistochemical analysis of CEACAM5 expression on the subject's tumor cells.

[0030] In certain embodiments, the subject has negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity ≧2+ in less than 1% of cells).

[0031] In certain embodiments, the subject has moderate CEACAM5 expression on tumor cells (intensity ≧2+ in greater than 1% and less than 50% of cells) as measured by immunohistochemistry.

[0032] In certain embodiments, the subject has high CEACAM5 expression on tumor cells (intensity ≧2+ in greater than or equal to 50% of cells) as measured by immunohistochemistry.

[0033] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 20 ng / mL.

[0034] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 25 ng / mL.

[0035] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 30 ng / mL.

[0036] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 35 ng / mL.

[0037] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 40 ng / mL.

[0038] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 45 ng / mL.

[0039] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 50 ng / mL.

[0040] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 55 ng / mL.

[0041] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 60 ng / mL.

[0042] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 61 ng / mL.

[0043] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 62 ng / mL.

[0044] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 63 ng / mL.

[0045] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 64 ng / mL.

[0046] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 65 ng / mL.

[0047] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 66 ng / mL.

[0048] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 67 ng / mL.

[0049] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 68 ng / mL.

[0050] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 69 ng / mL.

[0051] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 70 ng / mL.

[0052] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 71 ng / mL.

[0053] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 72 ng / mL.

[0054] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 73 ng / mL.

[0055] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 74 ng / mL.

[0056] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 75 ng / mL.

[0057] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 76 ng / mL.

[0058] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 77 ng / mL.

[0059] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 78 ng / mL.

[0060] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 79 ng / mL.

[0061] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 80 ng / mL.

[0062] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 85 ng / mL.

[0063] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 90 ng / mL.

[0064] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 95 ng / mL.

[0065] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 100 ng / mL.

[0066] In certain embodiments, tusamitamavutansine is administered at a dose of about 60 mg / m 2 ~about 190mg / m 2 (eg, about once every 2 or 3 weeks).

[0067] In certain embodiments, tusamitama ravtansine is administered about once every two weeks. In certain embodiments, tusamitama ravtansine is administered about once every three weeks. In certain embodiments, tusamitama ravtansine is administered about once every four weeks. In certain embodiments, tusamitama ravtansine is administered about once every five weeks. In certain embodiments, tusamitama ravtansine is administered about once every six weeks.

[0068] In certain embodiments, tusamitamavutansine is administered at a dose of about 60 mg / m 2 (eg, about once every 2 or 3 weeks).

[0069] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 (eg, about once every 2 or 3 weeks).

[0070] In certain embodiments, tusamitamavutansine is administered at about 120 mg / m 2 (eg, about once every 2 or 3 weeks).

[0071] In certain embodiments, tusamitama ravtansine is administered at a dose of about 150 mg / m 2 (eg, about once every 2 or 3 weeks).

[0072] In certain embodiments, tusamitamavutansine is administered at a dose of about 170 mg / m 2 (eg, about once every 2 or 3 weeks).

[0073] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 or more (e.g., 100 mg / m 2 ) administered approximately once every two weeks.

[0074] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 or more (e.g., 100 mg / m 2 ) administered approximately once every three weeks.

[0075] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 It is administered approximately once every two weeks at a dose of

[0076] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 It is administered approximately once every three weeks at a dose of

[0077] In certain embodiments, the methods further comprise administering to the subject an effective amount of at least one additional agent effective to treat cancer.

[0078] In certain embodiments, the additional agent is selected from the group consisting of immune checkpoint inhibitors (ICIs), platinum-based chemotherapy (e.g., cisplatin or carboplatin), pemetrexed, anti-VEGFR2, FOLFOX, FOLFIRI, TAS-102, anti-EGFR, and any combination thereof.

[0079] In certain embodiments, the ICI is an anti-PD-1 antibody.

[0080] In certain embodiments, the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, sintilimab, dostallimab, and ticerelizumab.

[0081] In certain embodiments, the anti-PD-1 antibody is pembrolizumab.

[0082] In certain embodiments, the ICI is an anti-PD-L1 antibody.

[0083] In certain embodiments, the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, durvalumab, emvafolimab, BMS-936559, CK-301, CS-1001, SHR-1316 (HTI-1088), CBT-502 (TQB-2450), and any combination thereof.

[0084] In certain embodiments, the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, and durvalumab.

[0085] In certain embodiments, the methods include administering to a subject an effective amount of tusamitamaravtansine and pembrolizumab.

[0086] In certain embodiments, the methods further comprise administering to the subject an effective amount of a platinum-based chemotherapy.

[0087] In certain embodiments, the platinum-based chemotherapy is selected from cisplatin and carboplatin.

[0088] In certain embodiments, the methods further comprise administering to the subject an effective amount of pemetrexed.

[0089] In certain embodiments, the methods include administering to a subject effective amounts of tusamitamaravtansine, pembrolizumab, and cisplatin.

[0090] In certain embodiments, the methods include administering to the subject effective amounts of tusamitamab ravtansine, pembrolizumab, cisplatin, and pemetrexed.

[0091] In certain embodiments, the methods include administering to a subject effective amounts of tusamitamaravtansine, pembrolizumab, and carboplatin.

[0092] In certain embodiments, the methods include administering to the subject effective amounts of tusamitamab ravtansine, pembrolizumab, carboplatin, and pemetrexed.

[0093] Another aspect of the present disclosure is a method of treating cancer in a subject, comprising: selecting a subject with cancer having a circulating carcinoembryonic antigen (CEA) of about 5 ng / mL or greater; administering to the subject an effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody; thereby treating cancer.

[0094] Another aspect of the disclosure is circulating carcinoembryonic antigen (CEA) for use as a biomarker in a method of treating cancer in a subject, the method comprising: selecting a subject with cancer having a circulating carcinoembryonic antigen (CEA) of about 5 ng / mL or greater; administering to the subject an effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody; thereby treating cancer.

[0095] In certain embodiments, the anti-CEACAM5 antibody is tusamitamab.

[0096] In certain embodiments, the ADC is tusamitamaravtansine.

[0097] In certain embodiments, the cancer is selected from the group consisting of colorectal cancer, gastric cancer, gastroesophageal junction cancer, esophageal cancer, lung cancer, cervical cancer, pancreatic cancer, ovarian cancer, thyroid cancer, bladder cancer, endometrial cancer, breast cancer, liver cancer, biliary tract cancer (e.g., cholangiocarcinoma), prostate cancer, and skin cancer.

[0098] In certain embodiments, the cancer is gastric cancer, gastroesophageal junction cancer, or esophageal cancer.

[0099] In certain embodiments, the cancer is gastric cancer.

[0100] In certain embodiments, the cancer is lung cancer.

[0101] In certain embodiments, the cancer is non-squamous non-small cell lung cancer (NSQ NSCLC).

[0102] In certain embodiments, the cancer is advanced or metastatic.

[0103] In certain embodiments, the NSQ NSCLC is advanced or metastatic.

[0104] In certain embodiments, circulating CEA is measured after performing immunohistochemical analysis of CEACAM5 expression on the subject's tumor cells.

[0105] In certain embodiments, the subject has negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity ≧2+ in less than 1% of cells).

[0106] In certain embodiments, the subject has moderate CEACAM5 expression on tumor cells (intensity ≧2+ in greater than 1% and less than 50% of cells) as measured by immunohistochemistry.

[0107] In certain embodiments, the subject has high CEACAM5 expression on tumor cells (intensity ≧2+ in greater than or equal to 50% of cells) as measured by immunohistochemistry.

[0108] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 20 ng / mL.

[0109] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 25 ng / mL.

[0110] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 30 ng / mL.

[0111] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 35 ng / mL.

[0112] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 40 ng / mL.

[0113] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 45 ng / mL.

[0114] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 50 ng / mL.

[0115] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 55 ng / mL.

[0116] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 60 ng / mL.

[0117] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 61 ng / mL.

[0118] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 62 ng / mL.

[0119] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 63 ng / mL.

[0120] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 64 ng / mL.

[0121] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 65 ng / mL.

[0122] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 66 ng / mL.

[0123] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 67 ng / mL.

[0124] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 68 ng / mL.

[0125] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 69 ng / mL.

[0126] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 70 ng / mL.

[0127] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 71 ng / mL.

[0128] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 72 ng / mL.

[0129] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 73 ng / mL.

[0130] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 74 ng / mL.

[0131] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 75 ng / mL.

[0132] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 76 ng / mL.

[0133] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 77 ng / mL.

[0134] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 78 ng / mL.

[0135] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 79 ng / mL.

[0136] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 80 ng / mL.

[0137] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 85 ng / mL.

[0138] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 90 ng / mL.

[0139] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 95 ng / mL.

[0140] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 100 ng / mL.

[0141] In certain embodiments, tusamitamavutansine is administered at a dose of about 60 mg / m 2 ~about 190mg / m 2 (eg, about once every 2 or 3 weeks).

[0142] In certain embodiments, tusamitama ravtansine is administered about once every two weeks. In certain embodiments, tusamitama ravtansine is administered about once every three weeks. In certain embodiments, tusamitama ravtansine is administered about once every four weeks. In certain embodiments, tusamitama ravtansine is administered about once every five weeks. In certain embodiments, tusamitama ravtansine is administered about once every six weeks.

[0143] In certain embodiments, tusamitamavutansine is administered at a dose of about 60 mg / m 2 (eg, about once every 2 or 3 weeks).

[0144] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 (eg, about once every 2 or 3 weeks).

[0145] In certain embodiments, tusamitamavutansine is administered at about 120 mg / m 2 (eg, about once every 2 or 3 weeks).

[0146] In certain embodiments, tusamitama ravtansine is administered at a dose of about 150 mg / m 2 (eg, about once every 2 or 3 weeks).

[0147] In certain embodiments, tusamitamavutansine is administered at a dose of about 170 mg / m 2 (eg, about once every 2 or 3 weeks).

[0148] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 or more (e.g., 100 mg / m 2 ) administered approximately once every two weeks.

[0149] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 or more (e.g., 100 mg / m 2 ) administered approximately once every three weeks.

[0150] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 It is administered approximately once every two weeks at a dose of

[0151] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 It is administered approximately once every three weeks at a dose of

[0152] In certain embodiments, the methods further comprise administering to the subject an effective amount of at least one additional agent effective to treat cancer.

[0153] In certain embodiments, the additional agent is selected from the group consisting of immune checkpoint inhibitors (ICIs), platinum-based chemotherapy (e.g., cisplatin or carboplatin), pemetrexed, anti-VEGFR2, FOLFOX, FOLFIRI, TAS-102, anti-EGFR, and any combination thereof.

[0154] In certain embodiments, the ICI is an anti-PD-1 antibody.

[0155] In certain embodiments, the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, sintilimab, dostallimab, and ticerelizumab.

[0156] In certain embodiments, the anti-PD-1 antibody is pembrolizumab.

[0157] In certain embodiments, the ICI is an anti-PD-L1 antibody.

[0158] In certain embodiments, the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, durvalumab, emvafolimab, BMS-936559, CK-301, CS-1001, SHR-1316 (HTI-1088), CBT-502 (TQB-2450), and any combination thereof.

[0159] In certain embodiments, the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, and durvalumab.

[0160] In certain embodiments, the methods include administering to a subject an effective amount of tusamitamaravtansine and pembrolizumab.

[0161] In certain embodiments, the methods further comprise administering to the subject an effective amount of a platinum-based chemotherapy.

[0162] In certain embodiments, the platinum-based chemotherapy is selected from cisplatin and carboplatin.

[0163] In certain embodiments, the methods further comprise administering to the subject an effective amount of pemetrexed.

[0164] In certain embodiments, the methods include administering to a subject effective amounts of tusamitamaravtansine, pembrolizumab, and cisplatin.

[0165] In certain embodiments, the methods include administering to the subject effective amounts of tusamitamab ravtansine, pembrolizumab, cisplatin, and pemetrexed.

[0166] In certain embodiments, the methods include administering to a subject effective amounts of tusamitamaravtansine, pembrolizumab, and carboplatin.

[0167] In certain embodiments, the methods include administering to the subject effective amounts of tusamitamab ravtansine, pembrolizumab, carboplatin, and pemetrexed.

[0168] Another aspect of the present disclosure is a method of treating cancer in a subject, comprising: Measuring first circulating carcinoembryonic antigen (CEA) in a subject in need of treatment for cancer; administering to the subject a first effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody if circulating CEA is initially measured in the subject at about 5 ng / mL or greater prior to administration; Measuring a second circulating CEA in the subject after the administering step; if the second circulating CEA measured in the subject is less than the first measured circulating CEA, administering to the subject a second effective amount of an ADC. thereby treating cancer.

[0169] Another aspect of the disclosure is circulating carcinoembryonic antigen (CEA) for use as a biomarker in a method of treating cancer in a subject, the method comprising: Measuring first circulating carcinoembryonic antigen (CEA) in a subject in need of treatment for cancer; administering to the subject a first effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody if circulating CEA is initially measured in the subject at about 5 ng / mL or greater prior to administration; Measuring a second circulating CEA in the subject after the administering step; if the second circulating CEA measured in the subject is less than the first measured circulating CEA, administering to the subject a second effective amount of an ADC. thereby treating cancer.

[0170] In certain embodiments, the anti-CEACAM5 antibody is tusamitamab.

[0171] In certain embodiments, the ADC is tusamitamaravtansine.

[0172] In certain embodiments, the cancer is selected from the group consisting of colorectal cancer, gastric cancer, gastroesophageal junction cancer, esophageal cancer, lung cancer, cervical cancer, pancreatic cancer, ovarian cancer, thyroid cancer, bladder cancer, endometrial cancer, breast cancer, liver cancer, biliary tract cancer (e.g., cholangiocarcinoma), prostate cancer, and skin cancer.

[0173] In certain embodiments, the cancer is gastric cancer.

[0174] In certain embodiments, the cancer is lung cancer.

[0175] In certain embodiments, the cancer is non-squamous non-small cell lung cancer (NSQ NSCLC).

[0176] In certain embodiments, the cancer is advanced or metastatic.

[0177] In certain embodiments, the NSQ NSCLC is advanced or metastatic.

[0178] In certain embodiments, circulating CEA is measured after performing immunohistochemical analysis of CEACAM5 expression on the subject's tumor cells.

[0179] In certain embodiments, the subject has negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity ≧2+ in less than 1% of cells).

[0180] In certain embodiments, the subject has moderate CEACAM5 expression on tumor cells (intensity ≧2+ in greater than 1% and less than 50% of cells) as measured by immunohistochemistry.

[0181] In certain embodiments, the subject has high CEACAM5 expression on tumor cells (intensity ≧2+ in greater than or equal to 50% of cells) as measured by immunohistochemistry.

[0182] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 20 ng / mL.

[0183] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 25 ng / mL.

[0184] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 30 ng / mL.

[0185] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 35 ng / mL.

[0186] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 40 ng / mL.

[0187] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 45 ng / mL.

[0188] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 50 ng / mL.

[0189] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 55 ng / mL.

[0190] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 60 ng / mL.

[0191] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 61 ng / mL.

[0192] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 62 ng / mL.

[0193] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 63 ng / mL.

[0194] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 64 ng / mL.

[0195] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 65 ng / mL.

[0196] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 66 ng / mL.

[0197] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 67 ng / mL.

[0198] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 68 ng / mL.

[0199] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 69 ng / mL.

[0200] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 70 ng / mL.

[0201] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 71 ng / mL.

[0202] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 72 ng / mL.

[0203] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 73 ng / mL.

[0204] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 74 ng / mL.

[0205] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 75 ng / mL.

[0206] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 76 ng / mL.

[0207] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 77 ng / mL.

[0208] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 78 ng / mL.

[0209] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 79 ng / mL.

[0210] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 80 ng / mL.

[0211] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 85 ng / mL.

[0212] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 90 ng / mL.

[0213] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 95 ng / mL.

[0214] In certain embodiments, prior to treatment, circulating CEA is greater than or equal to about 100 ng / mL.

[0215] In certain embodiments, tusamitamavutansine is administered at a dose of about 60 mg / m 2 ~about 190mg / m 2 (eg, about once every 2 or 3 weeks).

[0216] In certain embodiments, tusamitama ravtansine is administered about once every two weeks. In certain embodiments, tusamitama ravtansine is administered about once every three weeks. In certain embodiments, tusamitama ravtansine is administered about once every four weeks. In certain embodiments, tusamitama ravtansine is administered about once every five weeks. In certain embodiments, tusamitama ravtansine is administered about once every six weeks.

[0217] In certain embodiments, tusamitamavutansine is administered at a dose of about 60 mg / m 2 (eg, about once every 2 or 3 weeks).

[0218] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 (eg, about once every 2 or 3 weeks).

[0219] In certain embodiments, tusamitamavutansine is administered at about 120 mg / m 2 (eg, about once every 2 or 3 weeks).

[0220] In certain embodiments, tusamitama ravtansine is administered at a dose of about 150 mg / m 2 (eg, about once every 2 or 3 weeks).

[0221] In certain embodiments, tusamitamavutansine is administered at a dose of about 170 mg / m 2 (eg, about once every 2 or 3 weeks).

[0222] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 or more (e.g., 100 mg / m 2 ) administered approximately once every two weeks.

[0223] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 or more (e.g., 100 mg / m 2) administered approximately once every three weeks.

[0224] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 It is administered approximately once every two weeks at a dose of

[0225] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 It is administered approximately once every three weeks at a dose of

[0226] In certain embodiments, the methods further comprise administering to the subject an effective amount of at least one additional agent effective to treat cancer.

[0227] In certain embodiments, the additional agent is selected from the group consisting of immune checkpoint inhibitors (ICIs), platinum-based chemotherapy (e.g., cisplatin or carboplatin, pemetrexed, anti-VEGFR2, FOLFOX, FOLFIRI, TAS-102, anti-EGFR, and any combination thereof.

[0228] In certain embodiments, the ICI is an anti-PD-1 antibody.

[0229] In certain embodiments, the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, sintilimab, dostallimab, and ticerelizumab.

[0230] In certain embodiments, the anti-PD-1 antibody is pembrolizumab.

[0231] In certain embodiments, the ICI is an anti-PD-L1 antibody.

[0232] In certain embodiments, the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, durvalumab, emvafolimab, BMS-936559, CK-301, CS-1001, SHR-1316 (HTI-1088), CBT-502 (TQB-2450), and any combination thereof.

[0233] In certain embodiments, the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, and durvalumab.

[0234] In certain embodiments, the methods include administering to a subject an effective amount of tusamitamaravtansine and pembrolizumab.

[0235] In certain embodiments, the methods further comprise administering to the subject an effective amount of a platinum-based chemotherapy.

[0236] In certain embodiments, the platinum-based chemotherapy is selected from cisplatin and carboplatin.

[0237] In certain embodiments, the methods further comprise administering to the subject an effective amount of pemetrexed.

[0238] In certain embodiments, the methods include administering to a subject effective amounts of tusamitamaravtansine, pembrolizumab, and cisplatin.

[0239] In certain embodiments, the methods include administering to the subject effective amounts of tusamitamab ravtansine, pembrolizumab, cisplatin, and pemetrexed.

[0240] In certain embodiments, the methods include administering to a subject effective amounts of tusamitamaravtansine, pembrolizumab, and carboplatin.

[0241] In certain embodiments, the methods include administering to the subject effective amounts of tusamitamab ravtansine, pembrolizumab, carboplatin, and pemetrexed.

[0242] In embodiment 1, the disclosure relates to circulating carcinoembryonic antigen (CEA) for use as a biomarker in a method of treating cancer in a subject, the method comprising: Measuring circulating carcinoembryonic antigen (CEA) in a subject in need of treatment for cancer; administering to the subject an effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody if the subject has circulating CEA of about 5 ng / mL or greater; thereby treating cancer.

[0243] In embodiment 2, the disclosure relates to circulating carcinoembryonic antigen (CEA) for use as a biomarker in a method of treating cancer in a subject, the method comprising: selecting a subject with cancer having a circulating carcinoembryonic antigen (CEA) of about 5 ng / mL or greater; administering to the subject an effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody; thereby treating cancer.

[0244] In embodiment 3, the disclosure relates to circulating carcinoembryonic antigen (CEA) for use as a biomarker in a method of treating cancer in a subject, the method comprising: Measuring first circulating carcinoembryonic antigen (CEA) in a subject in need of treatment for cancer; administering to the subject a first effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody if circulating CEA is initially measured in the subject at about 5 ng / mL or greater prior to administration; Measuring a second circulating CEA in the subject after the administering step; if the second circulating CEA measured in the subject is less than the first measured circulating CEA, administering to the subject a second effective amount of an ADC. thereby treating cancer.

[0245] In embodiment 4, the present disclosure relates to an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent for use in a method of treating cancer in a subject in need thereof, The anti-CEACAM5 antibody comprises an HCDR1 having the amino acid sequence of SEQ ID NO:1, an HCDR2 having the amino acid sequence of SEQ ID NO:2, an HCDR3 having the amino acid sequence of SEQ ID NO:3, an LCDR1 having the amino acid sequence of SEQ ID NO:4, an LCDR2 having the amino acid sequence NTR, and an LCDR3 having the amino acid sequence of SEQ ID NO:5, the cytotoxic agent is a maytansinoid or a maytansinoid analog, and The method includes measuring circulating carcinoembryonic antigen (CEA) in the subject, and if circulating CEA in the subject is greater than or equal to about 5 ng / mL, administering to the subject an effective amount of the ADC, thereby treating cancer.

[0246] In embodiment 5, the present disclosure relates to the use of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent for the manufacture of a medicament for use in a method of treating cancer in a subject in need thereof, The anti-CEACAM5 antibody comprises an HCDR1 having the amino acid sequence of SEQ ID NO:1, an HCDR2 having the amino acid sequence of SEQ ID NO:2, an HCDR3 having the amino acid sequence of SEQ ID NO:3, an LCDR1 having the amino acid sequence of SEQ ID NO:4, an LCDR2 having the amino acid sequence NTR, and an LCDR3 having the amino acid sequence of SEQ ID NO:5, the cytotoxic agent is a maytansinoid or a maytansinoid analog, and The method includes measuring circulating carcinoembryonic antigen (CEA) in the subject, and if circulating CEA in the subject is greater than or equal to about 5 ng / mL, administering an effective amount of the pharmaceutical agent to the subject, thereby treating the cancer.

[0247] In embodiment 6, the present disclosure relates to a method of treating cancer in a subject in need thereof, the method comprising: measuring circulating carcinoembryonic antigen (CEA) in the subject; if the subject's circulating CEA is greater than or equal to about 5 ng / mL, administering to the subject an effective amount of an antibody-drug conjugate ADC, thereby treating the cancer. Including, The antibody-drug conjugate (ADC) comprises an anti-CEACAM5 antibody conjugated to a cytotoxic agent, The anti-CEACAM5 antibody comprises an HCDR1 having the amino acid sequence of SEQ ID NO:1, an HCDR2 having the amino acid sequence of SEQ ID NO:2, an HCDR3 having the amino acid sequence of SEQ ID NO:3, an LCDR1 having the amino acid sequence of SEQ ID NO:4, an LCDR2 having the amino acid sequence NTR, and an LCDR3 having the amino acid sequence of SEQ ID NO:5, and The cytotoxic agent is a maytansinoid or a maytansinoid analogue.

[0248] In embodiment 7, the present disclosure relates to any one of embodiments 1 to 6, wherein the cancer is selected from the group consisting of colorectal cancer, gastric cancer, gastroesophageal junction cancer, esophageal cancer, lung cancer, cervical cancer, pancreatic cancer, ovarian cancer, thyroid cancer, bladder cancer, endometrial cancer, breast cancer, liver cancer, biliary tract cancer (e.g., cholangiocarcinoma), prostate cancer, and skin cancer.

[0249] In embodiment 8, the present disclosure relates to any one of embodiments 1 to 7, wherein the cancer is gastric cancer.

[0250] In embodiment 9, the present disclosure relates to any one of embodiments 1 to 7, wherein the cancer is lung cancer.

[0251] In embodiment 10, the present disclosure relates to embodiment 9, wherein the cancer is non-squamous non-small cell lung cancer (NSQ NSCLC).

[0252] In embodiment 11, the present disclosure relates to embodiment 10, wherein the NSQ NSCLC is advanced or metastatic.

[0253] In embodiment 12, the present disclosure relates to any one of embodiments 1 to 11, wherein circulating CEA is measured after performing an immunohistochemical analysis of CEACAM5 expression on tumor cells of the subject.

[0254] In embodiment 13, the present disclosure relates to any one of embodiments 1 to 12, wherein the subject has negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in less than 1% of cells).

[0255] In embodiment 14, the present disclosure relates to any one of embodiments 1 to 12, wherein the subject has moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in more than 1% and less than 50% of cells).

[0256] In embodiment 15, the present disclosure relates to any one of embodiments 1 to 12, wherein the subject has high CEACAM5 expression on tumor cells (intensity of ≧2+ in ≧50% of cells) as measured by immunohistochemistry.

[0257] In embodiment 16, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 20 ng / mL.

[0258] In embodiment 17, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 25 ng / mL.

[0259] In embodiment 18, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 30 ng / mL.

[0260] In embodiment 19, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 35 ng / mL.

[0261] In embodiment 20, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA is greater than or equal to about 40 ng / mL before treatment.

[0262] In embodiment 21, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA is greater than or equal to about 45 ng / mL before treatment.

[0263] In embodiment 22, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 50 ng / mL.

[0264] In embodiment 23, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 55 ng / mL.

[0265] In embodiment 24, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 60 ng / mL.

[0266] In embodiment 25, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 61 ng / mL.

[0267] In embodiment 26, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 62 ng / mL.

[0268] In embodiment 27, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 63 ng / mL.

[0269] In embodiment 28, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 64 ng / mL.

[0270] In embodiment 29, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 65 ng / mL.

[0271] In embodiment 30, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 66 ng / mL.

[0272] In embodiment 31, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 67 ng / mL.

[0273] In embodiment 32, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 68 ng / mL.

[0274] In embodiment 33, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 69 ng / mL.

[0275] In embodiment 34, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 70 ng / mL.

[0276] In embodiment 35, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 71 ng / mL.

[0277] In embodiment 36, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 72 ng / mL.

[0278] In embodiment 37, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 73 ng / mL.

[0279] In embodiment 38, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 74 ng / mL.

[0280] In embodiment 39, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 75 ng / mL.

[0281] In embodiment 40, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 76 ng / mL.

[0282] In embodiment 41, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 77 ng / mL.

[0283] In embodiment 42, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 78 ng / mL.

[0284] In embodiment 43, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 79 ng / mL.

[0285] In embodiment 44, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 80 ng / mL.

[0286] In embodiment 45, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 80 ng / mL.

[0287] In embodiment 46, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 85 ng / mL.

[0288] In embodiment 47, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 90 ng / mL.

[0289] In embodiment 48, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 95 ng / mL.

[0290] In embodiment 49, the present disclosure relates to any one of embodiments 1 to 15, wherein circulating CEA before treatment is greater than or equal to about 100 ng / mL.

[0291] In embodiment 50, the present disclosure relates to any one of embodiments 1 to 49, wherein the anti-CEACAM5 antibody is tusamitamab.

[0292] In embodiment 51, the present disclosure relates to any one of embodiments 1 to 50, wherein the ADC is tusamitamaravtansine.

[0293] In embodiment 52, the present disclosure provides a method for administering tusamitamab ravtansine at a concentration of about 100 mg / m2 The above doses are administered about once every two weeks.

[0294] In embodiment 53, the present disclosure provides a method for administering tusamitamab ravtansine at a concentration of about 100 mg / m 2 The above doses are administered about once every three weeks.

[0295] In embodiment 54, the present disclosure relates to any one of embodiments 1 to 53, further comprising administering to the subject an effective amount of at least one additional agent effective to treat cancer.

[0296] In embodiment 55, the present disclosure relates to embodiment 54, wherein the additional agent is selected from the group consisting of immune checkpoint inhibitors (ICIs), platinum-based chemotherapy, pemetrexed, anti-VEGFR2, FOLFOX, FOLFIRI, TAS-102, anti-EGFR, and any combination thereof.

[0297] In embodiment 56, the present disclosure relates to embodiment 55, wherein the ICI is an anti-PD-1 antibody.

[0298] In embodiment 57, the present disclosure relates to embodiment 56, wherein the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, sintilimab, dostallimab, and tislelizumab.

[0299] In embodiment 58, the present disclosure relates to embodiment 56, in which the anti-PD-1 antibody is pembrolizumab.

[0300] In embodiment 59, the present disclosure relates to embodiment 55, wherein the ICI is an anti-PD-L1 antibody.

[0301] In embodiment 60, the present disclosure relates to embodiment 59, wherein the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, and durvalumab.

[0302] In embodiment 61, the present disclosure relates to any one of embodiments 1 to 58, comprising administering to a subject effective amounts of tusamitamaravtansine and pembrolizumab.

[0303] In embodiment 62, the present disclosure relates to embodiment 61, further comprising administering to the subject an effective amount of a platinum-based chemotherapeutic agent.

[0304] In embodiment 63, the present disclosure relates to embodiment 62, wherein the platinum-based chemotherapy is selected from cisplatin and carboplatin.

[0305] In embodiment 64, the present disclosure relates to embodiment 63, further comprising administering to the subject an effective amount of pemetrexed. [Brief description of the drawings]

[0306] [Figure 1] 1 is a graph showing circulating CEACAM5 at centrally assessed baseline and circulating CEA at locally assessed baseline according to an embodiment of the present teachings. [Diagram 2] FIG. 1 is a schematic diagram showing the study design. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0307] The present disclosure provides methods for identifying and treating patients having cancers that express CEACAM5, such as patients who would otherwise not be considered likely to respond to treatment due to low or moderate expression of CEACAM5 on tumor tissue, as measured by immunohistochemistry (IHC).

[0308] In accordance with the present disclosure, it has been discovered that patients with cancer and high levels of circulating CEA (e.g., greater than about 20 ng / mL, greater than about 50 ng / mL, greater than about 80 ng / mL, or greater than about 100 ng / mL) may be responsive to treatment despite having low or moderate expression of CEACAM5 on tumor tissue (≧2+ in less than 50% of tumor cells) as measured by immunohistochemistry.

[0309] In accordance with the present disclosure, it has been discovered that patients with cancer and high levels of circulating CEA (e.g., greater than about 20 ng / mL, greater than about 50 ng / mL, greater than about 80 ng / mL, or greater than about 100 ng / mL) may be responsive to treatments specifically directed against CEACAM5, despite having low or moderate expression of CEACAM5 on tumor tissue (≧2+ in less than 50% of tumor cells) as measured by immunohistochemistry.

[0310] Tusamitamab ravtansine (CAS Registry Number 2254086-60-5) is an immunoconjugate ADC that combines a humanized anti-CEACAM5 antibody (tusamitamab) with maytansinoid derivative 4 (DM4) [N2'-deacetyl-N2'-(4-methyl-4-mercapto-1-oxopentyl)-maytansine], a potent mitotic inhibitor that inhibits microtubule assembly. DM4 is stable in plasma and covalently attached to the antibody via an optimized linker SPDB [N-succinimidyl 4-(2-pyridyldithio)-butyrate] that is cleavable intracellularly. After binding and internalization in targeted cancer cells, the ADC is degraded, releasing the cytotoxic DM4 metabolites.

[0311] Tusamitama ravtansine specifically binds to the A3B3 domain of human CEACAM5 and does not recognize other CEACAMs that present A or / and B domains in their structure (CEACAM1, CEACAM6, CEACAM7 and CEACAM8). The naked antibody and ADC bind recombinant human CEACAM5 with an affinity of approximately 0.02 nM (ELISA) and show high affinity for CEACAM5-expressing tumor cells (K D APP Preliminary experiments indicate that tusamitamab ravtansine lacks effector activity.

[0312] After binding to the CEACAM5 antigen, tusamitamaravtansine is internalized by cancer cells via antigen-mediated endocytosis, delivered to lysosomes, and degraded to the lysine-linked derivative lysine-SPDB-DM4, which is further degraded in DM4 and subsequently S-methylated to form methyl-DM4 [Me-DM4], and all three metabolites have potent cytotoxic activity through binding to tubulin and inhibition of microtubule polymerization.

[0313] As used herein, high CEACAM5 cancer refers to several types of cancer, including colorectal cancer, lung cancer, gastric cancer, gastroesophageal junction cancer, esophageal cancer, lung cancer, uterine cancer, cervical cancer, pancreatic cancer, ovarian cancer, thyroid cancer, bladder cancer, endometrial cancer, breast cancer, liver cancer, prostate cancer and skin cancer. In some embodiments, the lung cancer is non-squamous non-small cell lung cancer. In some embodiments, high CEACAM5 expresser has an intensity of more than 2+ in at least 50% of expressing tumor cell population. High CEACAM5 expresser represents about 20% of lung cancer.

[0314] The ADC was analyzed in a Phase 1 / 2 study in heavily pretreated high CEACAM5 expressers for NSCLC. The ADC demonstrated competitive overall response rates (ORR) and duration of response (DoR). The most common adverse drug reactions (ADRs) were ocular toxicity (reversible without treatment discontinuation) and minimal hematological / neurological toxicity.

[0315] The majority of patients with NSCLC present with advanced stage disease at the time of diagnosis. The median overall survival (OS) for these patients is 8-12 months, with a 5-year survival rate of approximately 25% in 2015. Approximately 15%-20% of patients with NSCLC have tumors with significant genomic alterations amenable to targeted therapy, including epidermal growth factor receptor (EGFR) mutations and ROS receptor tyrosine kinase 1 (ROS1) and anaplastic lymphoma kinase (ALK) rearrangements.

[0316] Until recently, the only treatment option available for advanced or metastatic NSQ NSCLC lacking targetable mutations was chemotherapy. Systemic therapy with a platinum-based doublet regimen, with or without maintenance therapy, was the current first-line treatment for patients with advanced NSCLC.

[0317] More recently, immunotherapy has initiated a new paradigm for the treatment of NSCLC. For example, monoclonal antibodies targeting the programmed death-1 receptor (PD-1) / PD ligand-1 (PD-L1) pathway have emerged as a powerful new therapeutic tool in several clinical trials. Four drugs targeting the PD-1 pathway (nivolumab, pembrolizumab, atezolizumab, and cemiplimab) have been approved for the treatment of both chemotherapy-naïve and / or previously treated advanced-stage NSCLC, but only a small proportion of patients (20%-30%) respond to these therapies. Despite improved outcomes with these new lines of treatment, including anti-PD-1 / PD-L1 antibodies, the disease progresses in many cases.

[0318] The standard second-line treatment for NSCLC is docetaxel, whose activity has been shown to be enhanced by the addition of ramucirumab. Although there are other options available as monotherapy, such as pemetrexed or gemcitabine, there remains an unmet medical need for subsequent systemic treatment.

[0319] Non-small cell lung cancer is a disease in which malignant (cancer) cells form in the tissues of the lungs. Smoking is the main cause of the disease. It is a type of epithelial lung cancer other than small cell lung cancer. There are several types of non-small cell lung cancer. Each type of non-small cell lung cancer has different types of cancer cells. Each type of cancer cell grows and spreads in a different way. The types of non-small cell lung cancer are named for the type of cells found in the cancer and how the cells look under a microscope. (1) Squamous cell carcinoma: Cancer that begins in squamous cells (thin, flat cells that look like fish scales). This is also called epidermoid carcinoma. (2) Large cell carcinoma: Cancer that can begin in several types of larger cells. And (3) Adenocarcinoma: Cancer that begins in the cells that line the alveoli and make substances such as mucus.

[0320] As demonstrated by the recent approval of brentuximab vedotin for the treatment of Hodgkin's lymphoma and trastuzumab emtansine (T-DM1) for the treatment of recurrent metastatic HER2+ breast cancer, selective targeting of potent cytotoxic drugs to tumor cells using antibody-drug conjugates (ADCs) has now been shown to be an effective strategy for the treatment of cancer (Young A. et al., 2012 J Clin Oncol. 30(18):2183-9; Verma, S. et al., 2012 N Engl J Med. 19:1783-91). Many other malignant diseases with unmet medical needs could benefit from such treatment options. The mechanism of action of ADCs begins with binding to a specific antigen that is well expressed on tumor cells to achieve selective and efficient internalization of the drug.

[0321] Curative surgery (e.g., pneumonectomy, lobectomy, segmentectomy or wedge resection, sleeve resection) is the standard of care for patients with stage I NSCLC. Adjuvant treatment should only be offered as part of an investigational trial. Stage II and IIIA adjuvant cisplatin-based chemotherapy remains the gold standard for completely resected NSCLC tumors. Other chemotherapeutic agents used in combination with cisplatin or in combination with each other may include carboplatin, paclitaxel (TAXOL®), albumin-bound paclitaxel (nab-paclitaxel, ABRAXANE®), docetaxel (TAXOTERE®), gemcitabine (GEMZAR®), vinorelbine (NAVELBINE®), irinotecan (CAMPTOSAR®), etoposide (VP-16), vinblastine, and pemetrexed (ALIMTA®). Additionally, radiation therapy may be used for patients with N2 lymph nodes. For patients with advanced stage IIIB / IV or inoperable NSCLC, treatment may include multiple cycles of cisplatin-based chemotherapy plus third generation cytotoxic or cytostatic drugs (anti-EGFR, anti-VEGFR). (See Zarogoulidis et al., J Thorac Dis. 2013 Sep;5(Suppl4):S389-S396)

[0322] Treatment of cancer, including lung cancer, may include angiogenesis inhibitors, epidermal growth factor receptor (EGFR) inhibitors, and immune checkpoint inhibitors.

[0323] Angiogenesis inhibitors may include, but are not limited to, axitinib (INLYTA®), bevacizumab (AVASTIN®), cabozantinib (COMETRIQ®), everolimus (AFINITOR®, ZORTRESS®), lenalidomide (REVLIMID®), pazopanib (VOTRIENT®), ramucirumab (CYRAMZA®), regorafenib (STIVARGA®), sorafenib (NEXAVAR®), sunitinib (SUTENT®), thalidomide (SYNOVIR®, THALOMID®), vandetanib (CAPRELSA®), and Ziv-aflibercept (ZALTRAP®).

[0324] Epidermal growth factor receptor (EGFR) inhibitors can include, but are not limited to, gefitinib (IRESSA®), erlotinib (TARCEVA®), lapatinib (TYKERB®), cetuximab (ERBITUX®), neratinib (NERLYNX®), osimertinib (TAGRISSO®), panitumumab (VECTIBIX®), vandetanib (CAPRELSA®), necitumumab (PROTRAZZA®), and dacomitinib (VIZIMPRO®).

[0325] Immune checkpoint inhibitors include programmed death receptor 1 (PD-1) binders (e.g., pembrolizumab (KEYTRUDA®), nivolumab (OPDIVO®), cemiplimab (LIBTAYO®), programmed death-ligand 1 (PD-L1) binders (e.g., atezolizumab (TECENTRIQ®), avelumab (BAVENCIO®), durvalumab (IMFINZI®), CTLA-4 binders (e.g., ipilimumab (YERVOY®), OX40 or OX40L binders, adenosine A2A receptor binders, B7-H3 binders, B7-H4 These may include, but are not limited to, BTLA binders, indoleamine 2,3-dioxygenase binders, killer cell immunoglobulin-like receptor (KIR) binders, lymphocyte activation gene 3 (LAG-3) binders, nicotinamide adenine dinucleotide phosphate NADPH oxidase isoform (NOX2) binders, T cell immunoglobulin domain and mucin domain 3 (TIM-3) binders, T cell V domain Ig inhibitor of activation (VISTA) binders, glucocorticoid-induced TNFR family related gene (GITR) binders and sialic acid-binding immunoglobulin-type lectin 7 (SIGLEC7) binders.

[0326] CEACAM5 and its indications Carcinoembryonic antigen (CEA) is a glycoprotein involved in cell adhesion. CEA was first identified in 1965 as a protein normally expressed by fetal intestine during the first 6 months of pregnancy (Gold and Freedman, J Exp Med, 121, 439, 1965) and has been found in cancers of the pancreas, liver and colon. The CEA family belongs to the immunoglobulin superfamily. The CEA family, consisting of 18 genes, is subdivided into two subgroups of proteins: the carcinoembryonic antigen-related cell adhesion molecule (CEACAM) subgroup and the pregnancy-specific glycoprotein subgroup (Kammerer & Zimmermann, BMC Biology 2010, 8:12).

[0327] In humans, the CEACAM subgroup consists of seven members: CEACAM1, CEACAM3, CEACAM4, CEACAM5, CEACAM6, CEACAM7, CEACAM8. Numerous studies have shown that CEACAM5, identical to the first identified CEA, is highly expressed on the surface of colorectal, gastric, lung, breast, prostate, ovarian, cervical and bladder tumor cells and weakly expressed in a few normal epithelial tissues such as columnar and goblet cells of the colon, mucous cervical cells of the stomach and squamous cells of the esophagus and cervix (Hammarstroem et al, 2002, in "Tumor markers, Physiology, Pathobiology, Technology and Clinical Applications" Eds. Diamandis EP et al., AACC Press, Washington pp 375). CEACAM5 may therefore constitute a suitable therapeutic target for tumor-specific targeting approaches such as immunoconjugates. The extracellular domain of CEACAM family members is composed of repeated immunoglobulin-like (Ig-like) domains classified according to sequence homology into three types: A, B and N. CEACAM5 contains seven such domains, namely N, A1, B1, A2, B2, A3 and B3.

[0328] The CEACAM5 A1, A2 and A3 domains on the one hand and the B1, B2 and B3 domains on the other hand show high sequence homology, with the A domain of human CEACAM5 showing a pairwise sequence similarity of 84-87% and the B domain showing 69-80%. Furthermore, other human CEACAM members presenting A and / or B domains in their structure, namely CEACAM1, CEACAM6, CEACAM7 and CEACAM8, show homology with human CEACAM5. In particular, the A and B domains of the human CEACAM6 protein show sequence homology with either the A1 and A3 domains and the B1-B3 domains of human CEACAM5, respectively, which is even higher than that observed between the A and B domains of human CEACAM5.

[0329] Anti-CEACAM5 antibody: Considering the diagnostic or therapeutic purpose of targeting CEA, a number of anti-CEA antibodies have been generated. Specificity to related antigens has always been mentioned as a concern in the field, for example by Sharkey et al (1990, Cancer Research 50, 2823). Due to the above-mentioned homology, some of the previously described antibodies may demonstrate binding to repeated epitopes of CEACAM5 present in different immunoglobulin domains, may show cross-reactivity to other CEACAM members such as CEACAM1, CEACAM6, CEACAM7 or CEACAM8, and lack specificity for CEACAM5. The specificity of anti-CEACAM5 antibodies is desirable in terms of CEA-targeted therapy, as they bind to human CEACAM5-expressing tumor cells but not to some normal tissues expressing other CEACAM members. It is noteworthy that CEACAM1, CEACAM6 and CEACAM8 have been described to be expressed by neutrophils from humans and non-human primates (Ebrahimmnejad et al, 2000, Exp Cell Res, 260, 365; Zhao et al, 2004, J Immunol Methods 293, 207; Strickland et al, 2009 J Pathol, 218, 380) and have been shown to regulate granule formation and play a role in the immune response.

[0330] It has been shown that the ADC tusamitama ravtansine can be internalized into cells expressing CEACAM5 after binding and can induce cytotoxic activity against tumor cells in vitro. tusamitama ravtansine can also significantly inhibit tumor growth in vivo in mice bearing human primary colon and gastric tumors. See WO 2014 / 079886, which is incorporated herein in its entirety.

[0331] As used herein, the term "about" in a quantitative term refers to ±10% of the value it modifies (if the value is not divisible, such as the number of molecules or nucleotides, it is rounded up to the nearest whole number). For example, the phrase "about 100 mg" would encompass 90 mg to 110 mg, inclusive. The phrase "about 2500 mg" encompasses 2250 mg to 2750 mg. When applied to a percentage, the term "about" refers to plus or minus 10% to that percentage. For example, the phrase "about 20%" encompasses 18 to 22%, and "about 80%" encompasses 72 to 88%, inclusive. Furthermore, when "about" is used herein in conjunction with a quantitative term, it is understood that in addition to a value of plus or minus 10%, the exact value of the quantitative term is also contemplated and described. For example, the term "about 23%" expressly intends, describes, and includes exactly 23%.

[0332] It should be noted that the term "a" or "an" entity refers to one or more of that entity. For example, "symptoms" is understood to refer to one or more symptoms. Thus, the terms "a" (or "an"), "one or more," and "at least one" can be used interchangeably herein.

[0333] Furthermore, as used herein, "and / or" should be understood as specifically disclosing each of the two specified features or components with or without the other. Thus, as used herein in phrases such as "A and / or B," the term "and / or" is intended to include "A and B," "A or B," "A" (single) and "B" (single). Similarly, the term "and / or" when used in phrases such as "A, B and / or C" is intended to encompass each of the following embodiments: A, B and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A (single); B (single); and C (single).

[0334] Whenever an embodiment is described herein with the term "comprising," it is understood that other similar embodiments described with the terms "consisting of" and / or "consisting essentially of" are also provided.

[0335] As used herein, "CEACAM5" refers to "carcinoembryonic antigen-related cell adhesion molecule 5," also known as "CD66e" (cluster of differentiation 66e) or CEA. CEACAM5 is a glycoprotein involved in cell adhesion. CEACAM5 is highly expressed on the surface of, for example, colorectal cancer, gastric cancer, gastroesophageal junction cancer, esophageal cancer, lung and uterine tumor cells.

[0336] The reference sequence of full-length human CEACAM5, including the signal peptide (positions 1-34) and the propeptide (positions 686-702), is available from the GenBank database under the accession number AAA51967.1. Five nonsynonymous SNPs have been identified with a frequency of more than 2% in the Caucasian population, four of which are located in the N domain (positions 80, 83, 112, 113) and the last one in the A2 domain (position 398) of human CEACAM5.

[0337] Whenever an aspect or embodiment is described herein with the term "comprising," it is understood that other similar embodiments described with the terms "consisting of" and / or "consisting essentially of" are also provided.

[0338] The term "antibody" as used herein also includes antigen-binding fragments of an intact antibody molecule. As used herein, the terms "antigen-binding portion" of an antibody, "antigen-binding fragment" of an antibody, and the like include any naturally occurring, enzymatically derived, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex. Antigen-binding fragments of antibodies can be derived from intact antibody molecules using any suitable standard technique, such as, for example, proteolytic digestion or recombinant genetic engineering techniques, including the manipulation and expression of DNA encoding antibody variable domains and, optionally, constant domains. Such DNA is known and / or readily available, for example, from commercial sources, DNA libraries (including, for example, phage antibody libraries), or can be synthesized. The DNA can be sequenced and manipulated, for example, chemically or using molecular biology techniques, to place one or more variable and / or constant domains in the appropriate configuration or to introduce codons, create cysteine ​​residues, modify, add, or delete amino acids, and the like.

[0339] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single chain Fv (scFv) molecules; (vi) dAb fragments; (vii) a minimal recognition unit consisting of amino acid residues that mimic a hypervariable region of an antibody (e.g., an isolated complementarity determining region (CDR) such as a CDR3 peptide or a constraining FR3-CDR3-FR4 peptide. Other engineered molecules such as domain-specific antibodies, single domain antibodies, domain deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, VHHs or NANOBODYs (e.g., monovalent VHHs and bivalent VHHs), small modular immunopharmaceuticals (SMIPs) and shark variable IgNAR domains are also encompassed by the expression "antigen-binding fragment" as used herein.

[0340] Antigen-binding fragments of antibodies typically contain at least one variable domain. The variable domain may be of any size or amino acid composition and generally contains at least one CDR adjacent to or in frame with one or more framework sequences. In antigen-binding fragments having a VH domain associated with a VL domain, the VH domain and the VL domain may be positioned relative to each other in any suitable configuration. For example, the variable region may be dimeric and contain VH-VH, VH-VL or VL-VL dimers. Alternatively, the antigen-binding fragments of antibodies may contain monomeric VH or VL domains.

[0341] In certain embodiments, an antigen-binding fragment of an antibody may comprise at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains that may be found in an antigen-binding fragment of an antibody include: (i) VH-CH1; (ii) VH-CH2; (iii) VH-CH3; (iv) VH-CH1-CH2; (v) VH-CH1-CH2-CH3; (vi) VH-CH2-CH3; (vii) VH-CL; (viii) VL-CH1; (ix) VL-CH2; (x) VL-CH3; (xi) VL-CH1-CH2; (xii) VL-CH1-CH2-CH3; (xiii) VL-CH2-CH3; and (xiv) VL-CL. In any configuration of variable and constant domains, including any of the exemplary configurations listed above, the variable and constant domains may be directly linked to each other or may be linked by a complete or partial hinge or linker region. The hinge region may, in various embodiments, consist of at least two (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids that provide a flexible or semi-flexible linkage between adjacent variable and / or constant domains within a single polypeptide molecule. Furthermore, an antigen-binding fragment of an antibody may, in various embodiments, comprise a homodimer or heterodimer (or other multimer) of any of the variable and constant domain configurations listed above in non-covalent association with each other and / or with one or more monomeric VH or VL domains (e.g., by disulfide bonds).

[0342] In some embodiments, antibodies or antibody fragments for use in the methods of the present disclosure may be multispecific antibodies that may be specific for different epitopes of one target polypeptide or may include antigen binding domains specific for epitopes of two or more target polypeptides. An exemplary bispecific antibody format that can be used in connection with the present disclosure includes the use of a first immunoglobulin (Ig) CH3 domain and a second Ig CH3 domain, where the first and second Ig CH3 domains differ from each other in at least one amino acid, and the at least one amino acid difference reduces binding of the bispecific antibody to Protein A compared to a bispecific antibody lacking the amino acid difference. In one embodiment, the first Ig CH3 domain binds Protein A and the second Ig CH3 domain includes a mutation that reduces or eliminates Protein A binding, such as an H95R modification (according to IMGT exon numbering; H435R by EU numbering). The second CH3 may further include a Y96F modification (according to IMGT; Y436F by EU). Additional modifications that may be found within the second CH3 include: D16E, L18M, N44S, K52N, V57M and V82I for IgG1 antibodies (by IMGT; D356E, L358M, N384S, K392N, V397M and V422I by EU); N44S, K52N and V82I for IgG2 antibodies (by IMGT; N384S, K392N and V422I by EU); Q15R, N44S, K52N, V57M, R69K, E79Q and V82I for IgG4 antibodies (by IMGT; Q355R, N384S, K392N, V397M, R409K, E419Q and V422I by EU). Variations of the above bispecific antibody formats are contemplated within the scope of this disclosure. Any of the multispecific antibody formats, including the exemplary bispecific antibody formats disclosed herein, in various embodiments, can be adapted for use in the context of antigen-binding fragments of anti-CEACAM5 antibodies using routine techniques available in the art.

[0343] The CEACAM5 antibodies disclosed herein may contain one or more amino acid substitutions, insertions and / or deletions in the framework and / or CDR regions of the heavy and light chain variable domains compared to the corresponding germline sequences. Such mutations can be readily ascertained by comparing the amino acid sequences disclosed herein to germline sequences available, for example, from public antibody sequence databases. The present disclosure includes antibodies and antigen-binding fragments thereof derived from any of the amino acid sequences disclosed herein, in which one or more amino acids in one or more framework and / or CDR regions are backmutated to the corresponding germline residue or a conservative amino acid substitution (natural or non-natural) of the corresponding germline residue (such sequence changes are referred to herein as "germline backmutations"). Starting from the heavy and light chain variable region sequences disclosed herein, one skilled in the art can readily produce a large number of antibodies and antigen-binding fragments containing one or more individual germline backmutations or combinations thereof. In certain embodiments, all of the framework and / or CDR residues in the VH and / or VL domains are backmutated to the germline sequence. In other embodiments, only certain residues are mutated back to the germline sequence, for example only mutated residues found within the first 8 amino acids of FR1 or the last 8 amino acids of FR4, or only mutated residues found within CDR1, CDR2, or CDR3. Furthermore, the antibodies of the present disclosure may contain any combination of two or more germline reversions within the framework and / or CDR regions, i.e., certain individual residues are mutated back to the germline sequence, while certain other residues that differ from the germline sequence are maintained. Once obtained, antibodies and antigen-binding fragments containing one or more germline reversions can be readily tested for one or more desired properties, such as improved binding specificity, increased binding affinity, improved or enhanced antagonist or agonist biological properties (as the case may be), reduced immunogenicity, etc. Antibodies and antigen-binding fragments obtained in this general manner are encompassed within the present disclosure.

[0344] The constant region of an antibody is important in the ability of the antibody to fix complement and mediate cell-dependent cytotoxicity. Thus, an antibody isotype can be selected based on whether it is desirable for the antibody to mediate cytotoxicity.

[0345] The term "human antibody" as used herein is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Nevertheless, human antibodies featured in this disclosure may, in various embodiments, include amino acid residues, such as the CDRs and in some embodiments the CDR3, that are not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.

[0346] The term "recombinant human antibody" as used herein is intended to include all human antibodies that are prepared, expressed, created or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell (described further below), antibodies isolated from a recombinant combinatorial human antibody library (described further below), antibodies isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes (see, e.g., Taylor et al., (1992) Nucl. Acids Res. 20:6287-6295, incorporated herein by reference in its entirety), or antibodies prepared, expressed, created or isolated by any other means, including splicing of human immunoglobulin gene sequences into other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies have been subjected to in vitro mutagenesis (or in vivo somatic mutagenesis when using animals transgenic for human Ig sequences) such that the amino acid sequences of the VH and VL regions of the recombinant antibodies are derived from and related to human germline VH and VL sequences, but are sequences that may not naturally occur within the human antibody germline repertoire in vivo.

[0347] Human antibodies can exist in two forms related to hinge heterogeneity. In one embodiment, the immunoglobulin molecule comprises a stable four-chain construct of approximately 150-160 kDa in which the dimers are held together by interchain heavy chain disulfide bonds. In another embodiment, the dimers are not linked via interchain disulfide bonds and the approximately 75-80 kDa molecule is composed of covalently linked light and heavy chains (half antibodies). These embodiments / forms have been extremely difficult to separate, even after affinity purification.

[0348] The term "humanized antibody" refers to an antibody that is wholly or partially of non-human origin and that has been modified, for example by substituting certain amino acids in the framework regions of the VH and VL domains, to avoid or minimize an immune response in humans. The constant domains of a humanized antibody are most often human CH and CL domains.

[0349] Numerous methods for humanisation / humanisation of antibody sequences are known in the art; see for example the review by Almagro & Fransson (2008) Front Biosci. 13:1619-1633. One commonly used method is CDR grafting or antibody reshaping, which involves grafting the CDR sequences of a donor antibody, typically a murine antibody, onto the framework scaffold of a human antibody of different specificity. Since CDR grafting can reduce the binding specificity and affinity, and therefore the biological activity, of the CDR-grafted non-human antibody, back mutations can be introduced at selected positions of the CDR-grafted antibody to retain the binding specificity and affinity of the parent antibody. Identification of possible back mutation positions can be performed using information available in the literature and antibody databases. Amino acid residues that are candidates for back mutations are typically those located on the surface of the antibody molecule, whereas buried or less surface-exposed residues are usually not altered. An alternative humanization technique to CDR grafting and backmutation is resurfacing, in which non-surface exposed residues of non-human origin are retained and surface residues are changed to human residues. Another alternative technique, known as "guided selection" (Jespers et al. (1994) Biotechnology 12, 899), can be used to derive fully human antibodies from murine antibodies that preserve the epitope and binding specificity of the parent antibody.

[0350] The frequency of occurrence of the second form in various intact IgG isotypes is due to, but not limited to, structural differences associated with the hinge region isotype of the antibody. A single amino acid substitution in the hinge region of a human IgG4 hinge can significantly reduce the occurrence of the second form (Angal et al., (1993) Molecular Immunology 30:105, incorporated by reference in its entirety) to the level typically observed using a human IgG1 hinge. The present disclosure encompasses, in various embodiments, antibodies with one or more mutations in the hinge, CH2 or CH3 regions that may be desirable, for example, in production, to improve the yield of the desired antibody form.

[0351] As used herein, an "isolated antibody" refers to an antibody that has been identified and separated and / or recovered from at least one component of its natural environment. For example, an antibody that has been separated or removed from at least one component of an organism or tissue or cell in which it naturally occurs or is naturally produced is an "isolated antibody." In various embodiments, an isolated antibody also includes an antibody in situ within a recombinant cell. In other embodiments, an isolated antibody is an antibody that has been subjected to at least one purification or isolation step. In various embodiments, an isolated antibody may be substantially free of other cellular material and / or chemicals.

[0352] Terms such as "specifically bind" mean that an antibody or an antigen-binding fragment thereof forms a complex with an antigen that is relatively stable under physiological conditions. Methods for determining whether an antibody specifically binds to an antigen are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. For example, an antibody that "specifically binds" to CEACAM5 as used herein includes an antibody that binds to CEACAM5 or a portion thereof with a KD of less than about 1000 nM, less than about 500 nM, less than about 300 nM, less than about 200 nM, less than about 100 nM, less than about 90 nM, less than about 80 nM, less than about 70 nM, less than about 60 nM, less than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 4 nM, less than about 3 nM, less than about 2 nM, less than about 1 nM, or less than about 0.5 nM, as measured by a surface plasmon resonance assay. Specific binding is at least about 1×10 -6 In other embodiments, the dissociation constant is at least about 1×10 -7 M, 1×10 -8 M or 1×10 -9 An isolated antibody that specifically binds human CEACAM5 may, however, have cross-reactivity to other antigens, such as CEACAM5 molecules from other (non-human) species.

[0353] As used herein, the term "surface plasmon resonance" refers to an optical phenomenon that allows analysis of real-time interactions by detecting changes in protein concentration within a biosensor matrix, for example using a BIACORE™ system (Biacore Life Sciences division of GE Healthcare, Piscataway, NJ).

[0354] The term "KD," as used herein, is intended to refer to the equilibrium dissociation constant of an antibody-antigen interaction.

[0355] "Affinity" is theoretically defined by the equilibrium association between the whole antibody and the antigen. It can be experimentally evaluated by various known methods, such as measuring the association and dissociation rates by surface plasmon resonance or measuring the EC50 (or apparent KD) in immunochemical assays (ELISA, FACS). In these assays, EC50 is the concentration of antibody that induces a response halfway between the baseline and maximum after a specific exposure time to a defined concentration of antigen by ELISA (enzyme-linked immunosorbent assay) or cells expressing the antigen by FACS (fluorescence-activated cell sorting).

[0356] A monoclonal antibody that binds to antigen 1 (Ag1) is "cross-reactive" to antigen 2 (Ag2) if the EC50 is in the similar range for both antigens. In the present application, a monoclonal antibody that binds to Ag1 is cross-reactive to Ag2 if the ratio of the affinity of Ag2 to the affinity of Ag1 is less than or equal to 10 (e.g., 5, 2, 1 or 0.5), where affinity is measured in the same way for both antigens.

[0357] Affinity for human CEACAM5 or cynomolgus CEACAM5 can be determined as EC50 value in ELISA using soluble recombinant CEACAM5 as a capture antigen.

[0358] The antibodies of the disclosure may also have an apparent dissociation constant (apparent KD) of 25 nM or less, e.g., 20 nM or less, 10 nM or less, 5 nM or less, 3 nM or less, or 1 nM or less, as can be determined by FACS analysis on the tumor cell line MKN45 (DSMZ, ACC409) or patient-derived xenograft tumor cells (CR-IGR-034P available from Oncodesign Biotechnology, tumor collection CReMEC). D APP ) may be in the range of 0.01 to 20 nM, or may be in the range of 0.1 to 20 nM, 0.1 to 10 nM, or 0.1 to 5 nM.

[0359] Furthermore, antibodies according to the present disclosure have been shown to be capable of detecting CEACAM5 expression by immunohistochemistry in frozen and formalin-fixed and paraffin-embedded (FFPE) tissue sections.

[0360] The term "epitope" refers to an antigenic determinant that interacts with a specific antigen-binding site in the variable region of an antibody molecule known as the paratope. A single antigen may have more than one epitope. Thus, different antibodies may bind to different regions on an antigen and have different biological effects. Epitopes can be either conformational or linear. Conformational epitopes are generated by spatially juxtaposed amino acids from different segments of a linear polypeptide chain. Linear epitopes are those generated by adjacent amino acid residues in a polypeptide chain. In certain circumstances, epitopes may include portions of sugars, phosphoryl groups, or sulfonyl groups on an antigen.

[0361] Anti-CEACAM5 antibodies useful in the methods described herein may, in various embodiments, contain one or more amino acid substitutions, insertions, and / or deletions in the framework and / or CDR regions of the heavy and light chain variable domains compared to the corresponding germline sequences from which the antibody is derived. Such mutations can be readily ascertained by comparing the amino acid sequences disclosed herein to germline sequences available, for example, from public antibody sequence databases. The disclosure, in various embodiments, includes methods that include the use of antibodies and antigen-binding fragments thereof derived from any of the amino acid sequences disclosed herein, in which one or more amino acids in one or more framework and / or CDR regions are mutated to the corresponding residue in the germline sequence from which the antibody is derived or to the corresponding residue in another human germline sequence or to a conservative amino acid substitution of the corresponding germline residue (such sequence changes are collectively referred to herein as "germline mutations"). Numerous antibodies and antigen-binding fragments can be constructed that contain one or more individual germline mutations or combinations thereof. In certain embodiments, all of the framework and / or CDR residues in the VH and / or VL domains are mutated back to the residue found in the original germline sequence from which the antibody is derived. In other embodiments, only certain residues are mutated back to the original germline sequence, e.g., only mutated residues found within the first 8 amino acids of FR1 or the last 8 amino acids of FR4, or only mutated residues found within CDR1, CDR2, or CDR3. In other embodiments, one or more of the framework and / or CDR residues are mutated to the corresponding residue in a different germline sequence (i.e., a germline sequence different from the germline sequence from which the antibody was originally derived). Furthermore, an antibody may contain any combination of two or more germline mutations within the framework and / or CDR regions, e.g., certain individual residues are mutated to the corresponding residue in a particular germline sequence, while certain other residues that differ from the original germline sequence are maintained or mutated to the corresponding residue in a different germline sequence.Once obtained, antibodies and antigen-binding fragments containing one or more germline mutations can be readily tested for one or more desired properties, e.g., improved binding specificity, increased binding affinity, improved or enhanced antagonist or agonist biological properties (as the case may be), reduced immunogenicity, etc. Uses of antibodies and antigen-binding fragments obtained in this general manner are encompassed by the present disclosure.

[0362] The present disclosure also includes methods involving the use of anti-CEACAM5 antibodies that include variants of any of the HCVR, LCVR and / or CDR amino acid sequences disclosed herein having one or more conservative substitutions. For example, the present disclosure includes the use of anti-CEACAM5 antibodies that have HCVR, LCVR and / or CDR amino acid sequences that have, for example, 10 or less, 8 or less, 6 or less, 4 or less, etc., conservative amino acid substitutions compared to any of the HCVR, LCVR and / or CDR amino acid sequences disclosed herein.

[0363] According to the present disclosure, an anti-CEACAM5 antibody or antigen-binding fragment thereof, in various embodiments, comprises a heavy chain variable region (HCVR), a light chain variable region (LCVR) and / or a complementarity determining region (CDR) comprising any of the amino acid sequences of the anti-CEACAM5 antibodies described in WO 2014 / 079886, the entire contents of which are incorporated herein by reference.

[0364] Amino acid sequence modifications of the antibodies described herein are contemplated. For example, it may be desirable to improve the binding affinity and / or other biological properties of the antibody. It is known that when a humanized antibody is produced by simply grafting only the CDRs in the VH and VL of an antibody derived from a non-human animal into the FRs of the VH and VL of a human antibody, the antigen-binding activity is reduced compared to the original antibody derived from the non-human animal. It is believed that some amino acid residues in the VH and VL of the non-human antibody, not only in the CDRs but also in the FRs, may be directly or indirectly related to the antigen-binding activity. Therefore, when these amino acid residues are replaced with different amino acid residues from the FRs of the VH and VL of the human antibody, the binding activity is reduced. To solve this problem, an attempt must be made to identify amino acid residues in the amino acid sequences of the FRs of the VH and VL of the human antibody that are directly involved in the binding of the antibody, interact with the amino acid residues of the CDRs, or maintain the three-dimensional structure of the antibody and are directly involved in binding to the antigen in the human antibody with the non-human CDRs grafted therein. The reduction in antigen-binding activity could be increased by replacing the identified amino acids with amino acid residues of the original antibody derived from the non-human animal.

[0365] Modifications and variations can be made in the structure of the antibodies of the present disclosure and the DNA sequences encoding them and still result in a functional antibody or polypeptide possessing desirable characteristics.

[0366] A further object of the present disclosure also encompasses function-conserving variants of the polypeptides of the present disclosure. For example, certain amino acids can be substituted with other amino acids in the protein structure without any appreciable loss of activity. Because the interaction capacity and nature of a protein defines its biological functional activity, certain amino acid substitutions can be made in the protein sequence and, of course, in its DNA coding sequence, and still obtain a protein with similar properties. It is therefore believed that various changes can be made in the antibody sequences of the present disclosure or the corresponding DNA sequences encoding said polypeptides without appreciable loss of their biological activity. It is known in the art that certain amino acids can be substituted with other amino acids with similar hydropathic indexes or scores, resulting in proteins that still have similar biological activity, i.e., still obtain biologically functional equivalent proteins. In the antibodies or polypeptides of the present disclosure, it is also possible to use well-established techniques, such as the alanine scanning approach, to identify all amino acids that can be substituted without significant loss of binding to the antigen. Such residues can qualify as neutral, since they are not involved in antigen binding or maintaining the structure of the antibody. One or more of these neutral positions can be substituted with alanine or another amino acid without altering the main characteristics of the antibodies or polypeptides of the disclosure.

[0367] Neutral positions can be seen as positions where any amino acid substitution can be incorporated into the antibody. In fact, in the principle of alanine scanning, alanine is selected because this residue does not have any particular structural or chemical characteristics. It is generally accepted that if alanine can be substituted for a particular amino acid without changing the properties of the protein, then it is likely that many, if not all, other amino acid substitutions will also be neutral. In the opposite case, where alanine is the wild-type amino acid, if a particular substitution can be shown as neutral, it is likely that other substitutions will also be neutral. Thus, as outlined above, amino acid substitutions are generally based on the relative similarity of the amino acid side-chain substituents, such as their hydrophobicity, hydrophilicity, charge, size, etc. Exemplary substitutions that take into account any of the aforementioned characteristics are well known to those of skill in the art and include: arginine and lysine; glutamate and aspartate; serine and threonine; glutamine and asparagine; and valine, leucine and isoleucine.

[0368] It may also be desirable to modify the antibodies of the present disclosure with respect to effector function, for example to enhance antigen-dependent cell-mediated cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC) of the antibody. This can be achieved by introducing one or more amino acid substitutions into the Fc region of the antibody. Alternatively or additionally, cysteine ​​residues can be introduced into the Fc region, thereby allowing interchain disulfide bond formation in this region. The homodimeric antibody thus generated may have improved internalization capability and / or increased complement-mediated cell killing and / or antibody-dependent cellular cytotoxicity (ADCC) (Caron PC. et al. 1992; and Shopes B. 1992).

[0369] Another type of amino acid modification of the antibodies of the disclosure may be useful for altering the original glycosylation pattern of the antibody, i.e., by deleting one or more carbohydrate moieties found in the antibody and / or adding one or more glycosylation sites that are not present in the antibody. The presence of either of the tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, creates a potential glycosylation site. Addition or deletion of glycosylation sites to the antibody is conveniently accomplished by altering the amino acid sequence such that it contains one or more of the above tripeptide sequences (for N-linked glycosylation sites).

[0370] Another type of modification involves the removal of sequences identified in silico or experimentally as potentially contributing to degradation products or heterogeneity of antibody preparations. As an example, deamidation of asparagine and glutamine residues can occur depending on factors such as pH and surface exposure. Asparagine residues are primarily susceptible to deamidation when present in the sequence Asn-Gly, and less susceptible to deamidation in other dipeptide sequences such as Asn-Ala. Thus, when such deamidation sites, particularly Asn-Gly, are present in an antibody or polypeptide of the present disclosure, it may be desirable to remove the site, typically by conservative substitution, to remove one of the involved residues. Such substitutions in the sequence to remove one or more of the involved residues are also intended to be encompassed by the present disclosure.

[0371] Another type of covalent modification involves chemically or enzymatically coupling glycosides to the antibody. These procedures are advantageous in that they do not require production of the antibody in a host cell that has glycosylation capabilities for N- or O-linked glycosylation. Depending on the attachment mode used, the sugar(s) can be attached to (a) arginine and histidine, (b) free carboxyl groups, (c) free sulfhydryl groups such as those of cysteine, (d) free hydroxyl groups such as those of serine, threonine or hydroxyproline, (e) aromatic residues such as those of phenylalanine, tyrosine or tryptophan, or (f) the amide group of glutamine. For example, such methods are described in WO 87 / 05330.

[0372] Removal of any carbohydrate moieties present on an antibody can be accomplished chemically or enzymatically. Chemical deglycosylation requires exposure of the antibody to the compound trifluoromethanesulfonic acid, or an equivalent compound. This treatment results in the cleavage of most or all sugars except the linking sugar (N-acetylglucosamine or N-acetylgalactosamine), while leaving the antibody intact. Chemical deglycosylation has been described by Sojahr H. et al. (1987) and Edge, A. S. et al. (1981). Enzymatic cleavage of carbohydrate moieties on antibodies can be achieved by the use of various endo- and exoglycosidases as described by Thotakura, N. R. et al. (1987).

[0373] Another type of covalent modification of the antibody involves linking it to any of a variety of nonproteinaceous polymers, such as polyethylene glycol, polypropylene glycol, or polyoxyalkylenes, by the methods described in U.S. Pat. Nos. 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192; or 4,179,337.

[0374] In one embodiment, the anti-CEACAM5 antibody is tusamitamab (CAS Registry Number 2349294-95-5).

[0375] Tusamitamab comprises an HCDR1 having the amino acid sequence of SEQ ID NO: 1, an HCDR2 having the amino acid sequence of SEQ ID NO: 2, an HCDR3 having the amino acid sequence of SEQ ID NO: 3, an LCDR1 having the amino acid sequence of SEQ ID NO: 4, an LCDR2 having the amino acid sequence NTR, and an LCDR3 having the amino acid sequence of SEQ ID NO: 5. HCDR1 GFVFSSYD (SEQ ID NO: 1) HCDR2 ISSGGGIT (SEQ ID NO:2) HCDR3 AAHYFGSSGPFAY (SEQ ID NO: 3) LCDR1 ENIFSY (SEQ ID NO: 4) LCDR2 NTR LCDR3 QHHYGTPFT (SEQ ID NO:5)

[0376] Tusamitamab comprises a heavy chain variable domain (VH) having the amino acid sequence of SEQ ID NO:6 and a light chain variable domain (VL) having the amino acid sequence of SEQ ID NO:7. [ka]

[0377] Tusamitamab comprises a heavy chain (HC) having the amino acid sequence of SEQ ID NO:8 and a light chain (LC) having the amino acid sequence of SEQ ID NO:9. [ka]

[0378] Cytotoxic Payloads and Immunoconjugates The present disclosure also includes cytotoxic conjugates, or immunoconjugates, or antibody-drug conjugates, or conjugates. As used herein, these terms all have the same meaning and are interchangeable.

[0379] Accordingly, the present disclosure relates to an "immunoconjugate" comprising an antibody of the present disclosure (eg, an anti-CEACAM5 antibody) linked or conjugated to at least one growth inhibitory agent, such as a cytotoxic agent or a radioisotope.

[0380] "Growth inhibitory agent" or "anti-proliferative agent" may be used interchangeably and refer to a compound or composition that inhibits the growth of cells, especially tumor cells, either in vitro or in vivo.

[0381] The term "cytotoxic agent" as used herein refers to a substance that inhibits or prevents the function of a cell and / or causes destruction of a cell. The term "cytotoxic agent" is intended to include chemotherapeutic agents, enzymes, antibiotics, and toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant, or animal origin, fragments and / or variants thereof, as well as various antitumor or anticancer agents disclosed below. In some embodiments, the cytotoxic agent is a taxoid, a vinca, a maytansinoid or a maytansinoid analog, such as DM1 or DM4, a small drug, a tomaymycin or a pyrrolobenzodiazepine derivative, a cryptophycin derivative, a leptomycin derivative, an auristatin or a dolastatin analog, a prodrug, a topoisomerase II inhibitor, a DNA alkylating agent, an antitubulin agent, CC-1065 or a CC-1065 analog.

[0382] As used herein, "maytansinoids" refers to maytansinoids and maytansinoid analogs. Maytansinoids are drugs that inhibit microtubule formation and are highly toxic to mammalian cells.

[0383] Examples of suitable maytansinoids include maytansinol and maytansinol analogues.

[0384] Examples of suitable maytansinol analogues include those with modified aromatic rings and those with modifications at other positions.

[0385] Such suitable maytansinoids are described in U.S. Patent Nos. 4,424,219; 4,256,746; 4,294,757; 4,307,016; 4,313,946; 4,315,929; 4,331,598; 4,361,650; Nos. 4,362,663; 4,364,866; 4,450,254; 4,322,348; 4,371,533; 6,333,410; 5,475,092; 5,585,499; and 5,846,545.

[0386] Examples of suitable analogues of maytansinol having modified aromatic rings include the following: (1) C-19-dechloro (U.S. Pat. No. 4,256,746) (prepared by LAH reduction of ansamitocin P2); (2) C-20-hydroxy (or C-20-demethyl) + / - C-19-dechloro (U.S. Pat. Nos. 4,361,650 and 4,307,016) (prepared by demethylation with Streptomyces or Actinomyces or by dechlorination with LAH); and (3) C-20-demethoxy, C-20-acyloxy (-OCOR), + / -dechloro (U.S. Pat. No. 4,294,757) (prepared by acylation using acyl chlorides).

[0387] Examples of suitable analogues of maytansinol with modifications at other positions include the following: (1) C-9-SH (U.S. Pat. No. 4,424,219) (prepared by reaction of maytansinol with H2S or P2S5); (2) C-14-alkoxymethyl(demethoxy / CH2OR) (U.S. Pat. No. 4,331,598); (3) C-14-hydroxymethyl or acyloxymethyl (CHOH or CHOAc) (U.S. Pat. No. 4,450,254) (prepared from Nocardia); (4) C-15-hydroxy / acyloxy (U.S. Pat. No. 4,364,866) (prepared by conversion of maytansinol by Streptomyces); (5) C-15-methoxy (U.S. Pat. Nos. 4,313,946 and 4,315,929) (isolated from Trewia nudiflora); (6) C-18-N-demethyl (U.S. Pat. Nos. 4,362,663 and 4,322,348) (prepared by demethylation of maytansinol by Streptomyces); and (7) 4,5-deoxy (U.S. Pat. No. 4,371,533) (prepared by titanium trichloride / LAH reduction of maytansinol).

[0388] In one embodiment of the present disclosure, the cytotoxic conjugates of the present disclosure utilize as the cytotoxic agent a thiol-containing maytansinoid, formally referred to as N2'-deacetyl-N2'-(3-mercapto-1-oxopropyl)-maytansine (DM1), which is represented by the following structural formula (I): [ka]

[0389] In another embodiment, the cytotoxic conjugates of the present disclosure utilize as the cytotoxic agent the thiol-containing maytansinoid DM4, formally referred to as N2'-deacetyl-N-2'(4-methyl-4-mercapto-1-oxopentyl)-maytansine, which is represented by the following structural formula (II): [ka]

[0390] In further embodiments of the disclosure, other maytansines can be used, including thiol- and disulfide-containing maytansinoids with mono- or di-alkyl substitutions on the carbon atom bearing the sulfur atom. These include maytansinoids with acylated amino acid side chains with acyl groups bearing hindered sulfhydryl groups at C-3, C-14 hydroxymethyl, C-15 hydroxy, or C-20 desmethyl, where the carbon atom of the acyl group bearing the thiol functionality bears one or two substituents, said substituents being CH3, CH5, linear or branched alkyl or alkenyl with 1-10 groups and any aggregates that may exist in solution.

[0391] Examples of these cytotoxic agents and methods of conjugation are further provided in WO 2008 / 010101, which is incorporated by reference.

[0392] The term "radioisotope" means At 211 , Bi 212 , Er 169 , I 131 , I 125 , Y 90 , In 111 , P 32 , Re 186 , Re 188 , Sm 153 , Sr 89 The present disclosure is intended to include radioisotopes suitable for treating cancer, such as radioisotopes of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 112, 113, 114, 115, 120, 121, 136, 140, 141, 152, 163, 170, 175, 182, 196, 197, 198, 199,

[0393] Immunoconjugates according to the present disclosure can be prepared as described in WO 2004 / 091668, the contents of which are incorporated herein by reference in their entirety.

[0394] In some embodiments, the antibodies of the present disclosure are covalently attached to at least one growth inhibitory agent, either directly or via a cleavable or non-cleavable linker.

[0395] "Linker," as used herein, means a chemical moiety comprising a covalent bond or chain of atoms that covalently attaches a polypeptide (e.g., an antibody) to a drug (or prodrug) moiety. Suitable linkers are well known in the art and include disulfide groups, thioether groups, acid labile groups, photolabile groups, peptidase labile groups, and esterase labile groups. Exemplary linkers include, but are not limited to, heterobifunctional cross-linking reagents, such as N-succinimidyl pyridyldithiobutyrate (SPDB), butanoic acid 4-[(5-nitro-2-pyridinyl)dithio]-2,5-dioxo-1-pyrrolidinyl ester (nitro-SPDB), 4-(pyridin-2-yldisulfanyl)-2-sulfo-butyric acid (sulfo-SPDB), N-succinimidyl (2-pyridyldithio)propionate (SPDP), succinimidyl (N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC). , iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCL), active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bis-azido compounds (such as bis(p-azidobenzoyl)-hexanediamine), bis-diazonium derivatives (such as bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as toluene 2,6-diisocyanate) and bis-active fluorine compounds (such as 1,5-difluoro-2,4-dinitrobenzene). For example, ricin immunotoxins can be prepared as described in Vitetta et al (1987). Carbon-labeled 1-isothiocyanatobenzylmethyldiethylenetriaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for conjugation of radionucleotides to antibodies (WO 94 / 11026).

[0396] The linker may be a "cleavable linker" that facilitates the release of the cytotoxic or growth inhibitory agent in cells. For example, an acid-labile linker, a peptidase-sensitive linker, an esterase-labile linker, a photolabile linker, or a disulfide-containing linker (see, for example, U.S. Pat. No. 5,208,020) may be used. The linker may also be a "non-cleavable linker" (e.g., an SMCC linker), which may provide better resistance in some cases.

[0397] Alternatively, a fusion protein comprising an antibody of the disclosure and a cytotoxic or growth inhibitory polypeptide can be produced by recombinant techniques or peptide synthesis. The length of DNA can include respective regions encoding the two portions of the conjugate, either adjacent to each other or separated by a region encoding a linker peptide that does not destroy the desired properties of the conjugate.

[0398] The antibodies of the disclosure may also be used in dependent enzyme-mediated prodrug therapy by conjugating the polypeptide to a prodrug-activating enzyme that converts a prodrug (e.g., a peptidyl chemotherapeutic agent, see WO 81 / 01145) into an active anticancer drug (see, e.g., WO 88 / 07378 and U.S. Pat. No. 4,975,278). The enzyme component of the immunoconjugate useful for ADEPT includes any enzyme that can act on a prodrug to convert it into its more active, cytotoxic form. Enzymes useful in the methods of the disclosure include alkaline phosphatases, useful for converting phosphate-containing prodrugs to free drugs; arylsulfatases, useful for converting sulfate-containing prodrugs to free drugs; cytosine deaminases, useful for converting the non-toxic fluorocytosine to the anticancer drug, 5-fluorouracil; proteases, such as Serratia protease, thermolysin, subtilisin, carboxypeptidase, and cathepsins (e.g., cathepsins B and L), useful for converting peptide-containing prodrugs to free drugs; Examples of suitable enzymes include, but are not limited to, D-alanyl carboxypeptidase, useful for converting prodrugs to free drugs; carbohydrate cleaving enzymes such as O-galactosidase and neuraminidase, useful for converting glycosylated prodrugs to free drugs; P-lactamases, useful for converting P-lactam derivatized drugs to free drugs; and penicillin amidases, such as penicillin V amidase or penicillin G amidase, useful for converting drugs derivatized at their amine nitrogens bearing a phenoxyacetyl or phenylacetyl group, respectively, to free drugs. Enzymes can be covalently attached to the polypeptides of the present disclosure by techniques well known in the art, such as the use of heterobifunctional crosslinking reagents described above.

[0399] According to one embodiment, in the conjugate of the present disclosure, the growth inhibitory agent is a maytansinoid, in one embodiment DM1 or DM4.

[0400] In the conjugate, the antibody is conjugated to at least one growth inhibitory agent by a linking group, which in one embodiment is a cleavable or non-cleavable linker such as SPDB, sulfo-SPDB, or SMCC.

[0401] The conjugate may be selected from the group consisting of: Antibody-SPDB-DM4 conjugate of formula (III) [ka] ; Antibody-Sulfo-SPDB-DM4 Conjugate of Formula (IV) [ka] and Antibody-SMCC-DM1 conjugate of formula (V) [ka]

[0402] In one embodiment, the conjugate is a conjugate of formula (III), (IV) or (V) as defined above and the antibody is an antibody as described herein.

[0403] In formulas (III), (IV) and (V) above, "n" corresponds to the number of molecules of chemotherapeutic agent conjugated per antibody molecule. This corresponds to the "drug-to-antibody ratio" (or "DAR"), defined below, and can range from 1 to 10.

[0404] In one embodiment, the conjugate is tusamitamaravtansine (CAS Registry Number 2254086-60-5).

[0405] In general, the conjugates are (i) contacting an optionally buffered aqueous solution of an antibody according to the present disclosure with a solution of a linker and a cytotoxic compound; (ii) then, optionally, separating the conjugate formed in (i) from unreacted antibody, linker and cytotoxic compound; The composition can be obtained from a process including:

[0406] The aqueous solution of the cell-binding agent can be buffered with a buffer such as, for example, potassium phosphate, acetate, citrate, or N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES buffer). The buffer depends on the nature of the cell-binding agent. The cytotoxic compound is dissolved in an organic polar solvent, such as dimethylsulfoxide (DMSO) or dimethylacetamide (DMA).

[0407] The reaction temperature is usually comprised between 20 and 40°C. The reaction time can vary from 1 to 24 hours. The reaction between the cell-binding agent and the cytotoxic agent can be monitored by refractometry and / or size-exclusion chromatography (SEC) using a UV detector. If the conjugate yield is too low, the reaction time can be increased.

[0408] To carry out the separation of step (ii), several different chromatographic methods can be used by those skilled in the art. The conjugate can be purified, for example, by SEC, adsorption chromatography (ion exchange chromatography, IEC, etc.), hydrophobic interaction chromatography (HIC), affinity chromatography, mixed support chromatography such as hydroxyapatite chromatography, or high performance liquid chromatography (HPLC). Purification by dialysis or diafiltration can also be used.

[0409] As used herein, the term "aggregate" refers to an association that can be formed between two or more cell-binding agents, the agents being modified or not modified by conjugation. Aggregates can be formed under the influence of a number of parameters, such as high concentration of cell-binding agents in solution, pH of the solution, high shear force, the number of bound dimers and their hydrophobicity, temperature (see Wang & Gosh, 2008, J. Membrane Sci., 318:311-316 and references cited therein). It should be noted that the relative influence of some of these parameters has not been clearly established. In the case of proteins and antibodies, those skilled in the art are referred to Cromwell et al. (2006, AAPS Journal, 8(3):E572-E579). The content in aggregates can be determined using techniques well known to those skilled in the art, such as SEC (see Walter et al., 1993, Anal. Biochem., 212(2):469-480).

[0410] After step (i) or (ii), the conjugate-containing solution may be subjected to a further step (iii) of chromatography, ultrafiltration and / or diafiltration.

[0411] The conjugate is recovered at the end of these steps in aqueous solution.

[0412] According to one embodiment, a conjugate according to the present disclosure is characterized by a "drug-to-antibody ratio" (or "DAR") ranging from 1 to 10, such as from 2 to 5, such as from 3 to 4. This is generally the case for conjugates comprising a maytansinoid molecule.

[0413] This DAR number may vary depending on the nature of the antibody and drug (i.e. growth inhibitor) used together with the experimental conditions used for the conjugate (reaction time, nature of the solvent and nature of the co-solvent, if any, like the growth inhibitor / antibody ratio). Thus, contact between the antibody and the growth inhibitor results in a mixture containing several conjugates, which differ from each other by different drug-to-antibody ratios; optionally naked antibodies; optionally aggregates. The DAR determined is therefore an average value.

[0414] The method that can be used to determine the DAR is D and measuring the ratio of the absorbance of the solution of the substantially purified conjugate at 280 nm using a spectrophotometer. 280 nm is a wavelength commonly used to measure protein concentrations, such as antibody concentrations. D is selected to be able to distinguish the drug from the antibody, i.e., λ D is the drug has high absorbance and λ D λ is a wavelength that is sufficiently far from 280 nm to avoid substantial overlap of the drug and antibody absorbance peaks. D may be selected as 252 nm for maytansinoid molecules. Methods for DAR calculation can be obtained from Antony S. Dimitrov (ed), LLC, 2009, Therapeutic Antibodies and Protocols, vol 525, 445, Springer Science.

[0415] λ D (A λD ) and 280 nm (A 280 The absorbance of the conjugate at 100 nm is measured either by size exclusion chromatography (SEC) analysis of the monomer peak (allowing for the calculation of the "DAR(SEC)" parameter) or by a classical spectrophotometer device (allowing for the calculation of the "DAR(UV)" parameter). The absorbance can be expressed as: A λD =(c D xε DλD )+(cA xε AλD ) A 280 =(c D xε D280 )+(c A xε A280 ) During the ceremony, c D and c A are the concentrations of the drug and antibody in solution, respectively, ε DλD and ε D280 are λ D and the molar extinction coefficient of the drug at 280 nm, ε AλD and ε A280 are λ D and the molar extinction coefficient of the antibody at 280 nm.

[0416] Resolution of these two equations with two unknowns results in the following equation: c D =[(ε A280 xA λD )-(ε AλD xA 280 )] / [(ε DλD xε A280 )-(ε AλD xε D280 )] c A =[A 280 -(c D xε D280 )] / ε A280

[0417] The average DAR is then calculated from the ratio of the drug concentration to the antibody drug concentration. DAR=c D / c A .

[0418] Tsusami Tama Love Tanshin Tusamitamab ravtansine (CAS Registry Number 2254086-60-5) is an immunoconjugate ADC that combines a humanized anti-CEACAM5 antibody (tusamitamab) with maytansinoid derivative 4 (DM4) [N2-deacetyl-N2-(4-methyl-4-mercapto-1-oxopentyl)-maytansine], a potent mitotic inhibitor that inhibits microtubule assembly. DM4 is stable in plasma and covalently attached to the antibody via an optimized linker SPDB [N-succinimidyl 4-(2-pyridyldithio)-butyrate] that is cleavable intracellularly. After binding and internalization in targeted cancer cells, the ADC is degraded, releasing the cytotoxic DM4 metabolites.

[0419] The ADC tsusamitama ravtansine specifically binds to the A3B3 domain of human CEACAM5 and does not recognize other CEACAMs that present A or / and B domains in their structure (CEACAM1, CEACAM6, CEACAM7 and CEACAM8). The naked antibody and ADC bind recombinant human CEACAM5 with an affinity of approximately 0.02 nM (ELISA) and inhibit CEACAM5-expressing tumor cells (K D APP It exhibits high affinity for vasopressin (0.24-0.68 nM). Preliminary experiments indicate that vasopressin lacks effector activity.

[0420] After binding to the CEACAM5 antigen, tusamitamaravtansine is internalized by cancer cells via antigen-mediated endocytosis, delivered to lysosomes, and degraded to the lysine-linked derivative lysine-SPDB-DM4, which is further degraded in DM4 and subsequently S-methylated to form methyl-DM4 [Me-DM4], and all three metabolites have potent cytotoxic activity through binding to tubulin and inhibition of microtubule polymerization.

[0421] Pharmaceutical Compositions The antibodies or immunoconjugates of the disclosure can be combined with pharma- ceutically acceptable excipients, and optionally sustained release matrices (such as biodegradable polymers), to form therapeutic compositions.

[0422] Accordingly, another object of the present disclosure relates to a pharmaceutical composition comprising an antibody or immunoconjugate of the present disclosure and a pharma- ceutically acceptable carrier or excipient.

[0423] The present disclosure also relates to an antibody or immunoconjugate according to the present disclosure for use as a medicament.

[0424] The present disclosure also relates to an antibody, immunoconjugate, or compound according to the present disclosure for use in treating cancer.

[0425] "Pharmaceutical" or "Pharmaceutically acceptable" refers to molecular entities and compositions that do not produce adverse allergic or other untoward reactions when administered to a mammal, particularly a human, as appropriate. A pharmaceutically acceptable carrier or excipient refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type.

[0426] As used herein, a "pharmaceutical acceptable carrier" includes any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, etc. Examples of suitable carriers, diluents and / or excipients include water, amino acids, saline, phosphate buffered saline, phosphate buffer, acetate, citrate, succinate; amino acids and derivatives such as histidine, arginine, glycine, proline, glycylglycine, etc.; inorganic salts NaCl, calcium chloride; sugars or polyalcohols such as dextrose, glycerol, ethanol, sucrose, trehalose, mannitol; surfactants such as polysorbate 80, polysorbate 20, poloxamer 188, etc., and combinations thereof. In many cases, it is preferable to include an isotonic agent, such as a sugar, polyalcohol or sodium chloride, in the composition, and the formulation may also contain an antioxidant, such as tryptamine, and a stabilizer, such as Tween 20.

[0427] The form of the pharmaceutical composition, the route of administration, the dosage and the regimen will naturally depend on the condition to be treated, the severity of the disease, the age, weight and sex of the patient, etc.

[0428] The pharmaceutical compositions of the present disclosure may be formulated for topical, oral, parenteral, intranasal, intravenous, intramuscular, subcutaneous or intraocular administration, and the like.

[0429] In one embodiment, the pharmaceutical composition contains a pharma- ceutically acceptable vehicle for injectable preparations. These may be isotonic sterile saline (monosodium or disodium phosphate, sodium, potassium, calcium or magnesium chloride, etc., or mixtures of such salts) or a dry composition, in particular a lyophilized composition, which allows the constitution of an injectable solution upon addition of sterile water or saline, if appropriate.

[0430] The pharmaceutical composition may be administered via a drug combination device.

[0431] The dose used for administration can be adapted as a function of various parameters, for example as a function of the mode of administration used, the pathology involved or the duration of the treatment desired.

[0432] To prepare a pharmaceutical composition, an effective amount of the antibodies or immunoconjugates of the disclosure can be dissolved or dispersed in a pharma- ceutically acceptable carrier or aqueous medium.

[0433] Pharmaceutical forms suitable for injection use include sterile aqueous solutions or dispersions; formulations containing sesame oil, peanut oil or aqueous propylene glycol; sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and injectable in a suitable device or system for delivery without degradation. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi.

[0434] The solution of the active compound as a free base or a pharmacologically acceptable salt can be prepared in water mixed appropriately with a surfactant.Dispersions can also be prepared in glycerol, liquid polyethylene glycols and their mixtures and in oils.Under normal conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

[0435] The polypeptides, antibodies or immunoconjugates of the present disclosure can be formulated into compositions in neutral or salt form. Pharmaceutically acceptable salts include acid addition salts (formed with the free amino groups of the protein) and are formed with inorganic acids, such as hydrochloric or phosphoric acids, or organic acids, such as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases, such as sodium, potassium, ammonium, calcium, or ferric hydroxides, and organic bases, such as isopropylamine, trimethylamine, glycine, histidine, procaine, and the like.

[0436] The carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. Proper fluidity can be maintained, for example, by the use of a coating material such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. Prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it is preferable to include an isotonic agent, for example, sugar or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0437] Sterile injection solution is prepared by incorporating the required amount of active compound into a suitable solvent together with any other components listed above as necessary, followed by filtration sterilization.Generally, dispersion is prepared by incorporating various sterilized active components into a sterile vehicle that contains basic dispersion medium and other necessary components listed above.In the case of sterile powder for preparing sterile injection solution, the preferred preparation method is vacuum drying and freeze-drying technology, which obtains powder of active component + any additional desired component from its previously sterile filtered solution.

[0438] Preparation of more concentrated or highly concentrated solutions for direct injection is also contemplated, and the use of DMSO as a solvent is envisioned to result in extremely rapid penetration, delivering high concentrations of active agent to small tumor areas.

[0439] Upon formulation, solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective. The formulations are easily administered in a variety of dosage forms, such as the types of injectable solutions described above, although drug release capsules and the like can also be used.

[0440] For parenteral administration in aqueous solution, for example, the solution should be appropriately buffered if necessary, and the liquid diluent should first be rendered isotonic with sufficient saline or glucose. These aqueous solutions are suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration, among others. In this regard, the sterile aqueous media that can be used will be known to those skilled in the art in light of the present disclosure. For example, one dose can be dissolved in 1 ml of isotonic NaCl solution and added to 1000 ml of subcutaneous infusion fluid or injected at the proposed injection site (see, for example, "Remington's Pharmaceutical Sciences" 15th Edition, pages 1035-1038 and 1570-1580). Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject.

[0441] The antibodies or immunoconjugates of the disclosure may be formulated into a therapeutic mixture to contain about 0.01 to 100 milligrams per dose or the like.

[0442] In addition to antibodies or immunoconjugates formulated for parenteral administration, such as intravenous or intramuscular injection, other pharma- ceutically acceptable forms include, for example, tablets or other solids for oral administration; time-release capsules; and any other form currently in use.

[0443] In certain embodiments, the use of liposomes and / or nanoparticles is contemplated for the introduction of polypeptides into host cells. The formation and use of liposomes and / or nanoparticles are known to those of skill in the art.

[0444] Nanocapsules can generally entrap compounds in a stable and reproducible manner. To avoid side effects due to intracellular polymer overload, such ultrafine particles (size of about 0.1 μm) are generally designed using polymers that can be degraded in vivo. Biodegradable polyalkyl-cyanoacrylate nanoparticles or biodegradable polylactide or polylactide-co-glycolide nanoparticles that meet these requirements are contemplated for use in the present disclosure, and such particles can be easily made.

[0445] Liposomes are formed from phospholipids that disperse in aqueous media and spontaneously form multilamellar concentric bilayer vesicles, also called multilamellar vesicles (MLVs). MLVs generally have diameters between 25 nm and 4 μm. Sonication of MLVs results in the formation of small unilamellar vesicles (SUVs) with diameters ranging from 200 to 500 Å, which contain aqueous solution in their core. The physical properties of liposomes depend on pH, ionic strength and the presence of divalent cations.

[0446] Methods of administration and formulations The methods described herein include administering a therapeutically effective amount of an anti-CEACAM5 ADC to a subject. As used herein, an "effective amount" or a "therapeutically effective amount" is a dose of a therapeutic agent that results in the treatment of a CEACAM5-expressing cancer (e.g., lung cancer, gastric cancer, gastroesophageal junction cancer, or esophageal cancer). As used herein, "treating" refers to causing a detectable improvement in one or more symptoms associated with a CEACAM5-expressing cancer (e.g., lung cancer) or causing a biological effect (e.g., a reduction in the level of a particular biomarker) that correlates with the underlying pathological mechanism causing the condition or symptom. For example, a dose of an anti-CEACAM5 ADC that causes an improvement in any of the following symptoms or conditions associated with a CEACAM5-expressing cancer is considered a "therapeutically effective amount."

[0447] In another example, if a dose of an anti-CEACAM5 ADC does not result in a detectable improvement in one or more parameters or symptoms associated with a CEACAM5 expressing cancer (e.g., lung cancer, gastric cancer, gastroesophageal junction cancer or esophageal cancer) or does not cause a biological effect that correlates with the underlying pathological mechanism causing the cancerous condition or symptom, then the treatment was not effective.

[0448] According to some of these embodiments, the anti-CEACAM5 ADC is administered intravenously.

[0449] According to the methods of the disclosure, the therapeutically effective amount of an anti-CEACAM5 ADC administered to a subject will vary depending on the subject's age and size (e.g., body weight or body surface area), as well as the route of administration and other factors known to those of skill in the art.

[0450] In certain embodiments, the dose of the ADC varies depending on the body surface area of ​​the subject. In certain embodiments, the dose of the anti-CEACAM5 ADC administered to a subject is about 1 mg / m 2 ~about 500mg / m 2 In some embodiments, the dose of ADC administered to a subject is about 5 mg / m 2 ~about 300mg / m 2In various embodiments, the dose of ADC administered to a subject is about 5 mg / m 2 ~about 250mg / m 2 In various embodiments, the dose of the ADC administered to a subject is about 60 mg / m 2 ~about 190mg / m 2 It is.

[0451] In various embodiments, the dose is about 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, or 210 mg / m based on the subject's body surface area. 2 In certain embodiments, the dose of the ADC is about 100 mg / m 2 In certain embodiments, the dose of the ADC is about 150 mg / m 2 In certain embodiments, the dose of the ADC is about 170 mg / m 2 In certain embodiments, the dose of the ADC is about 190 mg / m 2 It is.

[0452] In various embodiments, the dose of the ADC is based on the subject's body surface area and is 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, or 210 mg / m 2 In certain embodiments, the dose of the ADC is 100 mg / m 2 In certain embodiments, the dose of the ADC is 150 mg / m 2 In certain embodiments, the dose of ADC is 170 mg / m 2 In certain embodiments, the dose of ADC is 190 mg / m 2 It is.

[0453] One aspect of the present disclosure is a method of treating cancer in a subject, comprising: Measuring circulating carcinoembryonic antigen (CEA) in a subject in need of treatment for cancer; administering to the subject an effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody if the subject has circulating CEA of about 5 ng / mL or greater; thereby treating cancer.

[0454] One aspect of the disclosure is circulating carcinoembryonic antigen (CEA) for use as a biomarker in a method of treating cancer in a subject, the method comprising: Measuring circulating carcinoembryonic antigen (CEA) in a subject in need of treatment for cancer; administering to the subject an effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody if the subject has circulating CEA of about 5 ng / mL or greater; thereby treating cancer.

[0455] One aspect of the disclosure is a method of diagnosing cancer in a subject, the method comprising at least: a) measuring in a biological sample the amount of circulating carcinoembryonic antigen (CEA) in said subject; b) comparing the measured amount obtained in step a) with a reference value, said reference value being 5 ng / mL; a) a measured amount of about 5 ng / mL or more indicative of cancer; Includes.

[0456] Circulating CEA can be measured in a sample selected from serum, plasma, and whole blood using any suitable method, such as enzyme-linked immunosorbent assay (ELISA). In certain embodiments, circulating CEA is measured in a serum sample using ELISA. In certain embodiments, circulating CEA is measured in a plasma sample using ELISA.

[0457] Circulating CEA can be measured at any time before, during or after treatment. In certain embodiments, circulating CEA is measured after performing immunohistochemical analysis of CEACAM5 expression on tumor cells of the subject. In certain embodiments, circulating CEA is first measured after performing immunohistochemical analysis of CEACAM5 expression on tumor cells of the subject.

[0458] In certain embodiments, the cancer is selected from the group consisting of colorectal cancer, gastric cancer, gastroesophageal junction cancer, esophageal cancer, lung cancer, cervical cancer, pancreatic cancer, ovarian cancer, thyroid cancer, bladder cancer, endometrial cancer, breast cancer, liver cancer, biliary tract cancer (e.g., cholangiocarcinoma), prostate cancer, and skin cancer.

[0459] In certain embodiments, the cancer is advanced or metastatic.

[0460] In certain embodiments, the cancer is gastric cancer.

[0461] In certain embodiments, the cancer is gastroesophageal junction cancer.

[0462] In certain embodiments, the cancer is esophageal cancer.

[0463] In certain embodiments, the cancer is lung cancer.

[0464] In certain embodiments, the cancer is small cell lung cancer (SCLC).

[0465] In certain embodiments, the cancer is non-small cell lung cancer (NSCLC).

[0466] In certain embodiments, the cancer is non-squamous non-small cell lung cancer (NSQ NSCLC).

[0467] In certain embodiments, the NSQ NSCLC is advanced or metastatic.

[0468] In certain embodiments, the subject has negative CEACAM5 expression on tumor cells (intensity ≧2+ in less than 1% of cells) as measured by immunohistochemistry.

[0469] In certain embodiments, the subject has moderate CEACAM5 expression on tumor cells (intensity ≧2+ in greater than 1% and less than 50% of cells) as measured by immunohistochemistry.

[0470] In certain embodiments, the subject has high CEACAM5 expression on tumor cells (intensity ≧2+ in greater than or equal to 50% of cells) as measured by immunohistochemistry.

[0471] In certain embodiments, prior to treatment, the subject's circulating CEA is about 5 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 10 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 15 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 20 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 25 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 30 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 35 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 40 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 45 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 50 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 55 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 60 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 61 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 62 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 63 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 64 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 65 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 66 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 67 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 68 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 69 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 70 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is greater than or equal to about 71 ng / mL. In certain embodiments, prior to treatment, the subject's circulating CEA is greater than or equal to about 72 ng / mL. In certain embodiments, prior to treatment, the subject's circulating CEA is greater than or equal to about 73 ng / mL.In certain embodiments, prior to treatment, the subject's circulating CEA is about 74 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 75 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 76 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 77 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 78 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 79 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 80 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 85 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 90 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 95 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 100 ng / mL or greater.

[0472] In one embodiment, the anti-CEACAM5 antibody is tusamitamab.

[0473] In certain embodiments, the ADC is tusamitamaravtansine.

[0474] In certain embodiments, tusamitamavutansine is administered at a dose of about 60 mg / m 2 ~about 190mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 60 mg / m 2 ~190mg / m 2 is administered at a dose of

[0475] In certain embodiments, tusamitamavutansine is administered at a dose of about 60 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 70 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 80 mg / m 2In certain embodiments, tusamitama ravtansine is administered at a dose of about 90 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 110 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 120 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 130 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 140 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 150 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 160 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 170 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 180 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 190 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 200 mg / m 2 is administered at a dose of

[0476] In certain embodiments, tusamitamavutansine is administered at a dose of 90 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 100 mg / m 2 In certain embodiments, tusamitamavutansine is administered at a dose of 110 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 120 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 130 mg / m 2In certain embodiments, tusamitama ravtansine is administered at a dose of 140 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 150 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 160 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 170 mg / m 2 In certain embodiments, tusamitamavutansine is administered at a dose of 180 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 190 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 200 mg / m 2 is administered at a dose of

[0477] In certain embodiments, tusamitama ravtansine is administered about once every two weeks. In certain embodiments, tusamitama ravtansine is administered about once every three weeks. In certain embodiments, tusamitama ravtansine is administered about once every four weeks. In certain embodiments, tusamitama ravtansine is administered about once every five weeks. In certain embodiments, tusamitama ravtansine is administered about once every six weeks.

[0478] In certain embodiments, tusamitama ravtansine is administered once every two weeks. In certain embodiments, tusamitama ravtansine is administered once every three weeks. In certain embodiments, tusamitama ravtansine is administered once every four weeks. In certain embodiments, tusamitama ravtansine is administered once every five weeks. In certain embodiments, tusamitama ravtansine is administered once every six weeks.

[0479] In certain embodiments, tusamitamavutansine is administered at a dose of about 100-200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of about 100-210 mg / m2 It is administered once every two weeks at a dose of

[0480] In certain embodiments, tusamitamavutansine is about 100-200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of 100 to 200 mg / m 2 It is administered once every two weeks at a dose of

[0481] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 100 mg / m 2 It is administered once every two weeks at a dose of

[0482] In certain embodiments, tusamitamavutansine is about 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 100 mg / m 2 It is administered once every two weeks at a dose of

[0483] In certain embodiments, tusamitamavutansine is administered at a dose of about 100-200 mg / m 2 In certain embodiments, tusamitamavutansine is administered at a dose of about 100-210 mg / m about once every three weeks. 2 It is administered once every three weeks at a dose of

[0484] In certain embodiments, tusamitamavutansine is administered at a dose of 100 to 200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of 100 to 200 mg / m 2 It is administered once every three weeks at a dose of

[0485] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 100 mg / m2 It is administered in doses of once every three weeks.

[0486] In certain embodiments, tusamitamavutansine is administered at a dose of 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 100 mg / m 2 It is administered once every three weeks at a dose of

[0487] In certain embodiments, tusamitamavutansine is administered at a dose of about 100-200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of about 100-200 mg / m 2 It is administered once every six weeks at a dose of

[0488] In certain embodiments, tusamitamavutansine is administered at a dose of 100 to 200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of 100-200 mg / m 2 It is administered once every six weeks at a dose of

[0489] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 100 mg / m 2 It is administered once every six weeks at a dose of

[0490] In certain embodiments, tusamitamavutansine is administered at a dose of 100 mg / m 2 In certain embodiments, tusamitamavutansine is administered at a dose of 100 mg / m 2 It is administered once every six weeks at a dose of

[0491] In certain embodiments, the methods further comprise administering to the subject an effective amount of at least one additional agent effective to treat cancer.

[0492] In certain embodiments, the additional agent is selected from the group consisting of immune checkpoint inhibitors (ICIs), platinum-based chemotherapy (e.g., cisplatin or carboplatin), pemetrexed, anti-VEGFR2, FOLFOX, FOLFIRI, TAS-102, anti-EGFR, and any combination thereof.

[0493] In certain embodiments, the ICI is an anti-PD-1 antibody.

[0494] In certain embodiments, the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, sintilimab, dostallimab, and ticerelizumab.

[0495] In certain embodiments, the anti-PD-1 antibody is pembrolizumab.

[0496] In certain embodiments, the ICI is an anti-PD-L1 antibody.

[0497] In certain embodiments, the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, durvalumab, emvafolimab, BMS-936559, CK-301, CS-1001, SHR-1316 (HTI-1088), CBT-502 (TQB-2450), and any combination thereof.

[0498] In certain embodiments, the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, and durvalumab.

[0499] In certain embodiments, the methods include administering to a subject an effective amount of tusamitamaravtansine and pembrolizumab.

[0500] In certain embodiments, the methods further comprise administering to the subject an effective amount of a platinum-based chemotherapy.

[0501] In certain embodiments, the platinum-based chemotherapy is selected from cisplatin and carboplatin.

[0502] In certain embodiments, the methods further comprise administering to the subject an effective amount of pemetrexed.

[0503] In certain embodiments, the methods include administering to a subject effective amounts of tusamitamaravtansine, pembrolizumab, and cisplatin.

[0504] In certain embodiments, the methods include administering to the subject effective amounts of tusamitamab ravtansine, pembrolizumab, cisplatin, and pemetrexed.

[0505] In certain embodiments, the methods include administering to a subject effective amounts of tusamitamaravtansine, pembrolizumab, and carboplatin.

[0506] In certain embodiments, the methods include administering to the subject effective amounts of tusamitamab ravtansine, pembrolizumab, carboplatin, and pemetrexed.

[0507] In certain embodiments, the additional agent is an anti-VEGFR2. In one embodiment, the anti-VEGFR2 is ramucirumab (CYRAMZA®).

[0508] In certain embodiments, the additional agent is FOLFOX, a chemotherapy regimen that includes folinic acid (leucovorin), fluorouracil (5-FU), and oxaliplatin.

[0509] In certain embodiments, the additional agent is FOLFIRI, a chemotherapy regimen that includes folinic acid (leucovorin), fluorouracil (5-FU), and irinotecan (ONIVYDE®).

[0510] In certain embodiments, the additional agent is TAS-102. TAS-102 is a combination of trifluridine and tipiracil hydrochloride.

[0511] In certain embodiments, the additional agent is an anti-EGFR, hi certain embodiments, the anti-EGFR is selected from cetuximab (ERBITUX®) and necitumumab (PORTRAZZA®).

[0512] In certain embodiments, the subject has moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in greater than 1% and less than 50% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamab ravtansine.

[0513] In certain embodiments, the subject has advanced or metastatic cancer, has moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in greater than 1% and less than 50% of cells), circulating CEA is greater than or equal to about 100 ng / mL, and the ADC is tusamitamaravtansine.

[0514] In certain embodiments, the subject has negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in less than 1% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamab ravtansine.

[0515] In certain embodiments, the subject has advanced or metastatic cancer, has negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in less than 1% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0516] In certain embodiments, the subject has NSQ NSCLC, moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in greater than 1% and less than 50% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0517] In certain embodiments, the subject has progressive or metastatic NSQ NSCLC, has moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in greater than 1% and less than 50% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0518] In certain embodiments, the subject has NSQ NSCLC, negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in less than 1% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0519] In certain embodiments, the subject has progressive or metastatic NSQ NSCLC, has negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in less than 1% of cells), and has circulating CEA of about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0520] In certain embodiments, the subject has progressive or metastatic NSQ NSCLC, has moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in greater than 1% and less than 50% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0521] One aspect of the present disclosure is a method of treating cancer in a subject, comprising: selecting a subject with cancer having a circulating carcinoembryonic antigen (CEA) of about 5 ng / mL or greater; administering to the subject an effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody; thereby treating cancer.

[0522] In one embodiment, the anti-CEACAM5 antibody is tusamitamab.

[0523] Another aspect of the disclosure is circulating carcinoembryonic antigen (CEA) for use as a biomarker in a method of treating cancer in a subject, the method comprising: selecting a subject with cancer having a circulating carcinoembryonic antigen (CEA) of about 5 ng / mL or greater; administering to the subject an effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody; thereby treating cancer.

[0524] Circulating CEA can be measured in a sample selected from serum, plasma, and whole blood using any suitable method, such as enzyme-linked immunosorbent assay (ELISA). In certain embodiments, circulating CEA is measured in a serum sample using ELISA. In certain embodiments, circulating CEA is measured in a plasma sample using ELISA.

[0525] Circulating CEA can be measured at any time before, during or after treatment. In certain embodiments, circulating CEA is measured after performing immunohistochemical analysis of CEACAM5 expression on tumor cells of the subject. In certain embodiments, circulating CEA is first measured after performing immunohistochemical analysis of CEACAM5 expression on tumor cells of the subject.

[0526] In certain embodiments, the cancer is selected from the group consisting of colorectal cancer, gastric cancer, gastroesophageal junction cancer, esophageal cancer, lung cancer, cervical cancer, pancreatic cancer, ovarian cancer, thyroid cancer, bladder cancer, endometrial cancer, breast cancer, liver cancer, biliary tract cancer (e.g., cholangiocarcinoma), prostate cancer, and skin cancer.

[0527] In certain embodiments, the cancer is advanced or metastatic.

[0528] In certain embodiments, the cancer is gastric cancer.

[0529] In certain embodiments, the cancer is lung cancer.

[0530] In certain embodiments, the cancer is small cell lung cancer (SCLC).

[0531] In certain embodiments, the cancer is non-small cell lung cancer (NSCLC).

[0532] In certain embodiments, the cancer is non-squamous non-small cell lung cancer (NSQ NSCLC).

[0533] In certain embodiments, the NSQ NSCLC is advanced or metastatic.

[0534] In certain embodiments, the subject has negative CEACAM5 expression on tumor cells (intensity ≧2+ in less than 1% of cells) as measured by immunohistochemistry.

[0535] In certain embodiments, the subject has moderate CEACAM5 expression on tumor cells (intensity ≧2+ in greater than 1% and less than 50% of cells) as measured by immunohistochemistry.

[0536] In certain embodiments, the subject has high CEACAM5 expression on tumor cells (intensity ≧2+ in greater than or equal to 50% of cells) as measured by immunohistochemistry.

[0537] In certain embodiments, prior to treatment, the subject's circulating CEA is about 5 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 10 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 15 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 20 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 25 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 30 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 35 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 40 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 45 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 50 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 55 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 60 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 61 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 62 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 63 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 64 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 65 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 66 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 67 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 68 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 69 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 70 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is greater than or equal to about 71 ng / mL. In certain embodiments, prior to treatment, the subject's circulating CEA is greater than or equal to about 72 ng / mL. In certain embodiments, prior to treatment, the subject's circulating CEA is greater than or equal to about 73 ng / mL.In certain embodiments, prior to treatment, the subject's circulating CEA is about 74 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 75 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 76 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 77 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 78 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 79 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 80 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 85 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 90 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 95 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 100 ng / mL or greater.

[0538] In certain embodiments, the ADC is tusamitamaravtansine.

[0539] In certain embodiments, tusamitamavutansine is administered at a dose of about 60 mg / m 2 ~about 190mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 60 mg / m 2 ~190mg / m 2 is administered at a dose of

[0540] In certain embodiments, tusamitamavutansine is administered at a dose of about 60 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 70 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 80 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 90 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 100 mg / m2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 110 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 120 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 130 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 140 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 150 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 160 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 170 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 180 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 190 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 200 mg / m 2 is administered at a dose of

[0541] In certain embodiments, tusamitamavutansine is administered at a dose of 90 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 100 mg / m 2 In certain embodiments, tusamitamavutansine is administered at a dose of 110 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 120 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 130 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 140 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 150 mg / m 2In certain embodiments, tusamitama ravtansine is administered at a dose of 160 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 170 mg / m 2 In certain embodiments, tusamitamavutansine is administered at a dose of 180 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 190 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 200 mg / m 2 is administered at a dose of

[0542] In certain embodiments, tusamitama ravtansine is administered about once every two weeks. In certain embodiments, tusamitama ravtansine is administered about once every three weeks. In certain embodiments, tusamitama ravtansine is administered about once every four weeks. In certain embodiments, tusamitama ravtansine is administered about once every five weeks. In certain embodiments, tusamitama ravtansine is administered about once every six weeks.

[0543] In certain embodiments, tusamitama ravtansine is administered once every two weeks. In certain embodiments, tusamitama ravtansine is administered once every three weeks. In certain embodiments, tusamitama ravtansine is administered once every four weeks. In certain embodiments, tusamitama ravtansine is administered once every five weeks. In certain embodiments, tusamitama ravtansine is administered once every six weeks.

[0544] In certain embodiments, tusamitamavutansine is administered at a dose of about 100-200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of about 100-200 mg / m 2 It is administered once every two weeks at a dose of

[0545] In certain embodiments, tusamitamavutansine is about 100-200 mg / m 2In certain embodiments, tusamitamab ravtansine is administered at a dose of 100 to 200 mg / m 2 It is administered once every two weeks at a dose of

[0546] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 100 mg / m 2 It is administered once every two weeks at a dose of

[0547] In certain embodiments, tusamitamavutansine is about 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 100 mg / m 2 It is administered once every two weeks at a dose of

[0548] In certain embodiments, tusamitamavutansine is administered at a dose of about 100-200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of about 100-200 mg / m 2 It is administered once every three weeks at a dose of

[0549] In certain embodiments, tusamitamavutansine is administered at a dose of 100 to 200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of 100 to 200 mg / m 2 It is administered once every three weeks at a dose of

[0550] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 100 mg / m 2 It is administered once every three weeks at a dose of

[0551] In certain embodiments, tusamitamavutansine is administered at a dose of 100 mg / m 2In certain embodiments, tusamitama ravtansine is administered at a dose of 100 mg / m 2 It is administered once every three weeks at a dose of

[0552] In certain embodiments, tusamitamavutansine is administered at a dose of about 100-200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of about 100-200 mg / m 2 It is administered once every six weeks at a dose of

[0553] In certain embodiments, tusamitamavutansine is administered at a dose of 100 to 200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of 100-200 mg / m 2 It is administered once every six weeks at a dose of

[0554] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 100 mg / m 2 It is administered once every six weeks at a dose of

[0555] In certain embodiments, tusamitamavutansine is administered at a dose of 100 mg / m 2 In certain embodiments, tusamitamavutansine is administered at a dose of 100 mg / m 2 It is administered once every six weeks at a dose of

[0556] In certain embodiments, tusamitamavutansine is administered at a dose of 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 100 mg / m 2 It is administered once every three weeks at a dose of

[0557] In certain embodiments, tusamitamavutansine is administered at a dose of 100 mg / m 2In certain embodiments, tusamitamavutansine is administered at a dose of 100 mg / m 2 It is administered once every six weeks at a dose of

[0558] In certain embodiments, the methods further comprise administering to the subject an effective amount of at least one additional agent effective to treat cancer.

[0559] In certain embodiments, the additional agent is selected from the group consisting of immune checkpoint inhibitors (ICIs), platinum-based chemotherapy (e.g., cisplatin or carboplatin), pemetrexed, anti-VEGFR2, FOLFOX, FOLFIRI, TAS-102, anti-EGFR, and any combination thereof.

[0560] In certain embodiments, the ICI is an anti-PD-1 antibody.

[0561] In certain embodiments, the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, sintilimab, dostallimab, and ticerelizumab.

[0562] In certain embodiments, the anti-PD-1 antibody is pembrolizumab.

[0563] In certain embodiments, the ICI is an anti-PD-L1 antibody.

[0564] In certain embodiments, the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, durvalumab, emvafolimab, BMS-936559, CK-301, CS-1001, SHR-1316 (HTI-1088), CBT-502 (TQB-2450), and any combination thereof.

[0565] In certain embodiments, the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, and durvalumab.

[0566] In certain embodiments, the methods include administering to a subject an effective amount of tusamitamaravtansine and pembrolizumab.

[0567] In certain embodiments, the methods further comprise administering to the subject an effective amount of a platinum-based chemotherapy.

[0568] In certain embodiments, the platinum-based chemotherapy is selected from cisplatin and carboplatin.

[0569] In certain embodiments, the methods further comprise administering to the subject an effective amount of pemetrexed.

[0570] In certain embodiments, the methods include administering to a subject effective amounts of tusamitamaravtansine, pembrolizumab, and cisplatin.

[0571] In certain embodiments, the methods include administering to the subject effective amounts of tusamitamab ravtansine, pembrolizumab, cisplatin, and pemetrexed.

[0572] In certain embodiments, the methods include administering to a subject effective amounts of tusamitamaravtansine, pembrolizumab, and carboplatin.

[0573] In certain embodiments, the methods include administering to the subject effective amounts of tusamitamab ravtansine, pembrolizumab, carboplatin, and pemetrexed.

[0574] In certain embodiments, the additional agent is an anti-VEGFR2. In one embodiment, the anti-VEGFR2 is ramucirumab (CYRAMZA®).

[0575] In certain embodiments, the additional agent is FOLFOX, a chemotherapy regimen that includes folinic acid (leucovorin), fluorouracil (5-FU), and oxaliplatin.

[0576] In certain embodiments, the additional agent is FOLFIRI, a chemotherapy regimen that includes folinic acid (leucovorin), fluorouracil (5-FU), and irinotecan (ONIVYDE®).

[0577] In certain embodiments, the additional agent is TAS-102. TAS-102 is a combination of trifluridine and tipiracil hydrochloride.

[0578] In certain embodiments, the additional agent is an anti-EGFR, hi certain embodiments, the anti-EGFR is selected from cetuximab (ERBITUX®) and necitumumab (PORTRAZZA®).

[0579] In certain embodiments, the subject has moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in greater than 1% and less than 50% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamab ravtansine.

[0580] In certain embodiments, the subject has advanced or metastatic cancer, has moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in greater than 1% and less than 50% of cells), circulating CEA is greater than or equal to about 100 ng / mL, and the ADC is tusamitamaravtansine.

[0581] In certain embodiments, the subject has negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in less than 1% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamab ravtansine.

[0582] In certain embodiments, the subject has advanced or metastatic cancer, has negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in less than 1% of cells), and circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0583] In certain embodiments, the subject has NSQ NSCLC, moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in greater than 1% and less than 50% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0584] In certain embodiments, the subject has progressive or metastatic NSQ NSCLC, has moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in greater than 1% and less than 50% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0585] In certain embodiments, the subject has NSQ NSCLC, negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in less than 1% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0586] In certain embodiments, the subject has progressive or metastatic NSQ NSCLC, has negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in less than 1% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0587] In certain embodiments, the subject has progressive or metastatic NSQ NSCLC, has moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in greater than 1% and less than 50% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0588] One aspect of the present disclosure is a method of treating cancer in a subject, comprising: Measuring first circulating carcinoembryonic antigen (CEA) in a subject in need of treatment for cancer; administering to the subject a first effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody if circulating CEA is initially measured in the subject at about 5 ng / mL or greater prior to administration; Measuring a second circulating CEA in the subject after the administering step; if the second circulating CEA measured in the subject is less than the first measured circulating CEA, administering to the subject a second effective amount of an ADC. thereby treating cancer.

[0589] Another aspect of the disclosure is circulating carcinoembryonic antigen (CEA) for use as a biomarker in a method of treating cancer in a subject, the method comprising: Measuring first circulating carcinoembryonic antigen (CEA) in a subject in need of treatment for cancer; administering to the subject a first effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody if circulating CEA is initially measured in the subject at about 5 ng / mL or greater prior to administration; Measuring a second circulating CEA in the subject after the administering step; if the second circulating CEA measured in the subject is less than the first measured circulating CEA, administering to the subject a second effective amount of an ADC. thereby treating cancer.

[0590] In one embodiment, the anti-CEACAM5 antibody is tusamitamab.

[0591] Circulating CEA can be measured in a sample selected from serum, plasma, and whole blood using any suitable method, such as enzyme-linked immunosorbent assay (ELISA). In certain embodiments, circulating CEA is measured in a serum sample using ELISA. In certain embodiments, circulating CEA is measured in a plasma sample using ELISA.

[0592] Circulating CEA can be measured at any time before, during or after treatment. In certain embodiments, circulating CEA is measured after performing immunohistochemical analysis of CEACAM5 expression on tumor cells of the subject. In certain embodiments, circulating CEA is first measured after performing immunohistochemical analysis of CEACAM5 expression on tumor cells of the subject.

[0593] In certain embodiments, the cancer is selected from the group consisting of colorectal cancer, gastric cancer, gastroesophageal junction cancer, esophageal cancer, lung cancer, cervical cancer, pancreatic cancer, ovarian cancer, thyroid cancer, bladder cancer, endometrial cancer, breast cancer, liver cancer, biliary tract cancer (e.g., cholangiocarcinoma), prostate cancer, and skin cancer.

[0594] In certain embodiments, the cancer is advanced or metastatic.

[0595] In certain embodiments, the cancer is gastric cancer.

[0596] In certain embodiments, the cancer is lung cancer.

[0597] In certain embodiments, the cancer is small cell lung cancer (SCLC).

[0598] In certain embodiments, the cancer is non-small cell lung cancer (NSCLC).

[0599] In certain embodiments, the cancer is non-squamous non-small cell lung cancer (NSQ NSCLC).

[0600] In certain embodiments, the NSQ NSCLC is advanced or metastatic.

[0601] In certain embodiments, the subject has negative CEACAM5 expression on tumor cells (intensity ≧2+ in less than 1% of cells) as measured by immunohistochemistry.

[0602] In certain embodiments, the subject has moderate CEACAM5 expression on tumor cells (intensity ≧2+ in greater than 1% and less than 50% of cells) as measured by immunohistochemistry.

[0603] In certain embodiments, the subject has high CEACAM5 expression on tumor cells (intensity ≧2+ in greater than or equal to 50% of cells) as measured by immunohistochemistry.

[0604] In certain embodiments, prior to treatment, the subject's circulating CEA is about 5 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 10 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 15 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 20 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 25 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 30 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 35 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 40 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 45 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 50 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 55 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 60 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 61 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 62 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 63 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 64 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 65 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 66 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 67 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 68 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 69 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 70 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is greater than or equal to about 71 ng / mL. In certain embodiments, prior to treatment, the subject's circulating CEA is greater than or equal to about 72 ng / mL. In certain embodiments, prior to treatment, the subject's circulating CEA is greater than or equal to about 73 ng / mL.In certain embodiments, prior to treatment, the subject's circulating CEA is about 74 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 75 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 76 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 77 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 78 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 79 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 80 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 85 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 90 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 95 ng / mL or greater. In certain embodiments, prior to treatment, the subject's circulating CEA is about 100 ng / mL or greater.

[0605] In one embodiment, the anti-CEACAM5 antibody is tusamitamab.

[0606] In certain embodiments, the ADC is tusamitamaravtansine.

[0607] In certain embodiments, tusamitamavutansine is administered at a dose of about 60 mg / m 2 ~about 190mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 60 mg / m 2 ~190mg / m 2 is administered at a dose of

[0608] In certain embodiments, tusamitamavutansine is administered at a dose of about 60 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 70 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 80 mg / m 2In certain embodiments, tusamitama ravtansine is administered at a dose of about 90 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 110 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 120 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 130 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 140 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 150 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 160 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 170 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 180 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 190 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 200 mg / m 2 is administered at a dose of

[0609] In certain embodiments, tusamitamavutansine is administered at a dose of 90 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 100 mg / m 2 In certain embodiments, tusamitamavutansine is administered at a dose of 110 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 120 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 130 mg / m 2In certain embodiments, tusamitama ravtansine is administered at a dose of 140 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 150 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 160 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 170 mg / m 2 In certain embodiments, tusamitamavutansine is administered at a dose of 180 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 190 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 200 mg / m 2 is administered at a dose of

[0610] In certain embodiments, tusamitama ravtansine is administered about once every two weeks. In certain embodiments, tusamitama ravtansine is administered about once every three weeks. In certain embodiments, tusamitama ravtansine is administered about once every four weeks. In certain embodiments, tusamitama ravtansine is administered about once every five weeks. In certain embodiments, tusamitama ravtansine is administered about once every six weeks.

[0611] In certain embodiments, tusamitama ravtansine is administered once every two weeks. In certain embodiments, tusamitama ravtansine is administered once every three weeks. In certain embodiments, tusamitama ravtansine is administered once every four weeks. In certain embodiments, tusamitama ravtansine is administered once every five weeks. In certain embodiments, tusamitama ravtansine is administered once every six weeks.

[0612] In certain embodiments, tusamitamavutansine is administered at a dose of about 100-200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of about 100-200 mg / m2 It is administered once every two weeks at a dose of

[0613] In certain embodiments, tusamitamavutansine is about 100-200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of 100 to 200 mg / m 2 It is administered once every two weeks at a dose of

[0614] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 100 mg / m 2 It is administered once every two weeks at a dose of

[0615] In certain embodiments, tusamitamavutansine is about 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 100 mg / m 2 It is administered once every two weeks at a dose of

[0616] In certain embodiments, tusamitamavutansine is administered at a dose of about 100-200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of about 100-200 mg / m 2 It is administered once every three weeks at a dose of

[0617] In certain embodiments, tusamitamavutansine is administered at a dose of 100 to 200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of 100 to 200 mg / m 2 It is administered once every three weeks at a dose of

[0618] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 100 mg / m2 It is administered once every three weeks at a dose of

[0619] In certain embodiments, tusamitamavutansine is administered at a dose of 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of 100 mg / m 2 It is administered once every three weeks at a dose of

[0620] In certain embodiments, tusamitamavutansine is administered at a dose of about 100-200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of about 100-200 mg / m 2 It is administered once every six weeks at a dose of

[0621] In certain embodiments, tusamitamavutansine is administered at a dose of 100 to 200 mg / m 2 In certain embodiments, tusamitamab ravtansine is administered at a dose of 100-200 mg / m 2 It is administered once every six weeks at a dose of

[0622] In certain embodiments, tusamitamavutansine is administered at a dose of about 100 mg / m 2 In certain embodiments, tusamitama ravtansine is administered at a dose of about 100 mg / m 2 It is administered once every six weeks at a dose of

[0623] In certain embodiments, tusamitamavutansine is administered at a dose of 100 mg / m 2 In certain embodiments, tusamitamavutansine is administered at a dose of 100 mg / m 2 It is administered once every six weeks at a dose of

[0624] In certain embodiments, the methods further comprise administering to the subject an effective amount of at least one additional agent effective to treat cancer.

[0625] In certain embodiments, the additional agent is selected from the group consisting of immune checkpoint inhibitors (ICIs), platinum-based chemotherapy (e.g., cisplatin or carboplatin), pemetrexed, anti-VEGFR2, FOLFOX, FOLFIRI, TAS-102, anti-EGFR, and any combination thereof.

[0626] In certain embodiments, the ICI is an anti-PD-1 antibody.

[0627] In certain embodiments, the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, sintilimab, dostallimab, and ticerelizumab.

[0628] In certain embodiments, the anti-PD-1 antibody is pembrolizumab.

[0629] In certain embodiments, the ICI is an anti-PD-L1 antibody.

[0630] In certain embodiments, the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, durvalumab, emvafolimab, BMS-936559, CK-301, CS-1001, SHR-1316 (HTI-1088), CBT-502 (TQB-2450), and any combination thereof.

[0631] In certain embodiments, the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, and durvalumab.

[0632] In certain embodiments, the methods include administering to a subject an effective amount of tusamitamaravtansine and pembrolizumab.

[0633] In certain embodiments, the methods further comprise administering to the subject an effective amount of a platinum-based chemotherapy.

[0634] In certain embodiments, the platinum-based chemotherapy is selected from cisplatin and carboplatin.

[0635] In certain embodiments, the methods further comprise administering to the subject an effective amount of pemetrexed.

[0636] In certain embodiments, the methods include administering to a subject effective amounts of tusamitamaravtansine, pembrolizumab, and cisplatin.

[0637] In certain embodiments, the methods include administering to the subject effective amounts of tusamitamab ravtansine, pembrolizumab, cisplatin, and pemetrexed.

[0638] In certain embodiments, the methods include administering to a subject effective amounts of tusamitamaravtansine, pembrolizumab, and carboplatin.

[0639] In certain embodiments, the methods include administering to the subject effective amounts of tusamitamab ravtansine, pembrolizumab, carboplatin, and pemetrexed.

[0640] In certain embodiments, the additional agent is an anti-VEGFR2. In one embodiment, the anti-VEGFR2 is ramucirumab (CYRAMZA®).

[0641] In certain embodiments, the additional agent is FOLFOX, a chemotherapy regimen that includes folinic acid (leucovorin), fluorouracil (5-FU), and oxaliplatin.

[0642] In certain embodiments, the additional agent is FOLFIRI, a chemotherapy regimen that includes folinic acid (leucovorin), fluorouracil (5-FU), and irinotecan (ONIVYDE®).

[0643] In certain embodiments, the additional agent is TAS-102. TAS-102 is a combination of trifluridine and tipiracil hydrochloride.

[0644] In certain embodiments, the additional agent is an anti-EGFR, hi certain embodiments, the anti-EGFR is selected from cetuximab (ERBITUX®) and necitumumab (PORTRAZZA®).

[0645] In certain embodiments, the subject has moderate CEACAM5 expression on tumor cells (intensity of ≧2+ in greater than 1% and less than 50% of cells) as measured by immunohistochemistry, circulating CEA of about 100 ng / mL or greater, and the ADC is tusamitamab ravtansine.

[0646] In certain embodiments, the subject has advanced or metastatic cancer, has moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in greater than 1% and less than 50% of cells), circulating CEA is greater than or equal to about 100 ng / mL, and the ADC is tusamitamaravtansine.

[0647] In certain embodiments, the subject has negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in less than 1% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamab ravtansine.

[0648] In certain embodiments, the subject has advanced or metastatic cancer, has negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in less than 1% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0649] In certain embodiments, the subject has NSQ NSCLC, moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in greater than 1% and less than 50% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0650] In certain embodiments, the subject has progressive or metastatic NSQ NSCLC, has moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in greater than 1% and less than 50% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0651] In certain embodiments, the subject has NSQ NSCLC, negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in less than 1% of cells), circulating CEA is about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0652] In certain embodiments, the subject has progressive or metastatic NSQ NSCLC, has negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in less than 1% of cells), has circulating CEA of about 100 ng / mL or greater, and the ADC is tusamitamaravtansine.

[0653] Various embodiments of the present disclosure In embodiment 1, the disclosure relates to circulating carcinoembryonic antigen (CEA) for use as a biomarker in a method of treating cancer in a subject, the method comprising: Measuring circulating carcinoembryonic antigen (CEA) in a subject in need of treatment for cancer; administering to the subject an effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody if the subject has circulating CEA of about 5 ng / mL or greater; thereby treating cancer.

[0654] In embodiment 2, the disclosure relates to circulating carcinoembryonic antigen (CEA) for use as a biomarker in a method of treating cancer in a subject, the method comprising: selecting a subject with cancer having a circulating carcinoembryonic antigen (CEA) of about 5 ng / mL or greater; administering to the subject an effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody; thereby treating cancer.

[0655] In embodiment 3, the disclosure relates to circulating carcinoembryonic antigen (CEA) for use as a biomarker in a method of treating cancer in a subject, the method comprising: Measuring first circulating carcinoembryonic antigen (CEA) in a subject in need of treatment for cancer; administering to the subject a first effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody if circulating CEA is initially measured in the subject at about 5 ng / mL or greater prior to administration; Measuring a second circulating CEA in the subject after the administering step; if the second circulating CEA measured in the subject is less than the first measured circulating CEA, administering to the subject a second effective amount of an ADC. thereby treating cancer.

[0656] In embodiment 4, the present disclosure relates to a circulating carcinoembryonic antigen (CEA) of any one of embodiments 1 to 3, wherein the anti-CEACAM5 antibody comprises an HCDR1 having the amino acid sequence of SEQ ID NO: 1, an HCDR2 having the amino acid sequence of SEQ ID NO: 2, an HCDR3 having the amino acid sequence of SEQ ID NO: 3, an LCDR1 having the amino acid sequence of SEQ ID NO: 4, an LCDR2 having the amino acid sequence NTR, and an LCDR3 having the amino acid sequence of SEQ ID NO: 5.

[0657] In embodiment 5, the present disclosure relates to the circulating carcinoembryonic antigen (CEA) of any one of embodiments 1 to 4, wherein the cytotoxic agent is a maytansinoid or a maytansinoid analog.

[0658] In embodiment 6, the present disclosure relates to an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent for use in a method of treating cancer in a subject in need thereof, The anti-CEACAM5 antibody comprises an HCDR1 having the amino acid sequence of SEQ ID NO:1, an HCDR2 having the amino acid sequence of SEQ ID NO:2, an HCDR3 having the amino acid sequence of SEQ ID NO:3, an LCDR1 having the amino acid sequence of SEQ ID NO:4, an LCDR2 having the amino acid sequence NTR, and an LCDR3 having the amino acid sequence of SEQ ID NO:5, the cytotoxic agent is a maytansinoid or a maytansinoid analog, and The method includes measuring circulating carcinoembryonic antigen (CEA) in the subject, and if circulating CEA in the subject is greater than or equal to about 5 ng / mL, administering to the subject an effective amount of the ADC, thereby treating cancer.

[0659] In embodiment 7, the present disclosure relates to the use of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent for the manufacture of a medicament for use in a method of treating cancer in a subject in need thereof, The anti-CEACAM5 antibody comprises an HCDR1 having the amino acid sequence of SEQ ID NO:1, an HCDR2 having the amino acid sequence of SEQ ID NO:2, an HCDR3 having the amino acid sequence of SEQ ID NO:3, an LCDR1 having the amino acid sequence of SEQ ID NO:4, an LCDR2 having the amino acid sequence NTR, and an LCDR3 having the amino acid sequence of SEQ ID NO:5, the cytotoxic agent is a maytansinoid or a maytansinoid analog, and The method includes measuring circulating carcinoembryonic antigen (CEA) in the subject, and if circulating CEA in the subject is greater than or equal to about 5 ng / mL, administering an effective amount of the pharmaceutical agent to the subject, thereby treating the cancer.

[0660] In embodiment 8, the present disclosure relates to a method of treating cancer in a subject in need thereof, the method comprising: measuring circulating carcinoembryonic antigen (CEA) in the subject; if the subject's circulating CEA is greater than or equal to about 5 ng / mL, administering to the subject an effective amount of an antibody-drug conjugate ADC, thereby treating the cancer. Including, The antibody-drug conjugate (ADC) comprises an anti-CEACAM5 antibody conjugated to a cytotoxic agent, The anti-CEACAM5 antibody comprises an HCDR1 having the amino acid sequence of SEQ ID NO:1, an HCDR2 having the amino acid sequence of SEQ ID NO:2, an HCDR3 having the amino acid sequence of SEQ ID NO:3, an LCDR1 having the amino acid sequence of SEQ ID NO:4, an LCDR2 having the amino acid sequence NTR, and an LCDR3 having the amino acid sequence of SEQ ID NO:5, and The cytotoxic agent is a maytansinoid or a maytansinoid analogue.

[0661] In embodiment 9, the present disclosure relates to any one of embodiments 4 to 8, wherein the method further comprises: selecting a subject with cancer having a circulating carcinoembryonic antigen (CEA) of about 5 ng / mL or greater; administering to the subject an effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody; thereby treating cancer.

[0662] In embodiment 10, the present disclosure relates to any one of embodiments 4 to 8, wherein the method further comprises: Measuring first circulating carcinoembryonic antigen (CEA) in a subject in need of treatment for cancer; administering to the subject a first effective amount of an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody if circulating CEA is initially measured in the subject at about 5 ng / mL or greater prior to administration; Measuring a second circulating CEA in the subject after the administering step; if the second circulating CEA measured in the subject is less than the first measured circulating CEA, administering to the subject a second effective amount of an ADC. thereby treating cancer.

[0663] In embodiment 11, the present disclosure relates to any one of embodiments 1 to 10, wherein the cancer is selected from the group consisting of colorectal cancer, gastric cancer, gastroesophageal junction cancer, esophageal cancer, lung cancer, cervical cancer, pancreatic cancer, ovarian cancer, thyroid cancer, bladder cancer, endometrial cancer, breast cancer, liver cancer, biliary tract cancer (e.g., cholangiocarcinoma), prostate cancer, and skin cancer.

[0664] In embodiment 12, the present disclosure relates to any one of embodiments 1 to 11, wherein the cancer is gastric cancer.

[0665] In embodiment 13, the present disclosure relates to any one of embodiments 1 to 12, wherein the cancer is lung cancer.

[0666] In embodiment 14, the present disclosure relates to any one of embodiments 1 to 13, wherein circulating CEA is measured after performing immunohistochemical analysis of CEACAM5 expression on tumor cells of the subject.

[0667] In embodiment 15, the present disclosure relates to a method for treating a subject having a tumor, the method comprising the steps of: (a) determining whether a subject has a tumor that is resistant to CEACAM5 expression; and (b) determining whether a subject has a tumor that is resistant to CEACAM5 expression. In embodiment 15, the present disclosure relates to a method for treating a subject having a tumor that is resistant to CEACAM5 expression; The subject has moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity ≧2+ in greater than 1% and less than 50% of cells); or Any one of embodiments 1-14, wherein the subject has high CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in ≧50% of cells).

[0668] In embodiment 16, the present disclosure relates to a method for treating a patient with a circulating CEA of about 20 ng / mL or greater prior to treatment; Prior to treatment, circulating CEA was greater than or equal to about 50 ng / mL, or Prior to treatment, circulating CEA was greater than or equal to about 80 ng / mL, or Any one of embodiments 1-15, wherein prior to treatment, circulating CEA is greater than or equal to about 100 ng / mL.

[0669] In embodiment 17, the present disclosure relates to any one of embodiments 1 to 16, wherein the anti-CEACAM5 antibody is tusamitamab.

[0670] In embodiment 18, the present disclosure relates to any one of embodiments 1 to 17, wherein the ADC is tusamitamaravtansine.

[0671] In embodiment 19, the present disclosure provides a method for administering tusamitamab ravtansine at a concentration of about 100 mg / m 2 or more once every 2 weeks, or tusamitamab ravtansine is administered at a dose of about 100 mg / m 2 The above doses are administered about once every three weeks.

[0672] In embodiment 20, the present disclosure relates to any one of embodiments 1 to 19, further comprising administering to the subject an effective amount of at least one additional agent effective to treat cancer.

[0673] In embodiment 21, the present disclosure relates to embodiment 20, wherein the additional agent is selected from the group consisting of immune checkpoint inhibitors (ICIs), platinum-based chemotherapy, pemetrexed, anti-VEGFR2, FOLFOX, FOLFIRI, TAS-102, anti-EGFR, and any combination thereof.

[0674] In embodiment 22, the present disclosure relates to embodiment 21, wherein the ICI is an anti-PD-1 antibody or an anti-PD-L1 antibody.

[0675] In embodiment 23, the present disclosure relates to embodiment 22, wherein the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, sintilimab, dostallimab, and tislelizumab.

[0676] In embodiment 24, the present disclosure relates to embodiment 22, wherein the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, and durvalumab.

[0677] In embodiment 25, the present disclosure relates to any one of embodiments 1 to 23, comprising administering to the subject effective amounts of tusamitamaravtansine and pembrolizumab.

[0678] In embodiment 26, the present disclosure relates to embodiment 25, further comprising administering to the subject an effective amount of a platinum-based chemotherapeutic agent.

[0679] In embodiment 27, the present disclosure relates to embodiment 26, wherein the platinum-based chemotherapy is selected from cisplatin and carboplatin.

[0680] In embodiment 28, the present disclosure relates to embodiment 27, further comprising administering to the subject an effective amount of pemetrexed. EXAMPLES

[0681] Example 1. Background and study rationale In the first-in-human (FIH) TED13751 study, a cohort of 64 participants with heavily pretreated NSQ NSCLC with high CEACAM5 expression (defined as intensity ≥2+ in at least 50% of tumor cells) using an IHC method applied in prescreening on archived tumor samples was treated with 100 mg / m 2Patients were treated with tusamitamazavvtansine Q2W, which has shown promising antitumor activity. This antitumor activity was associated with an overall response rate of 20.3% (13 participants had PR) per Response Evaluation Criteria in Solid Tumors (RECIST) 1.1 (95% CI: 12.27% to 31.71%), warranting further development of tusamitamazavvvtansine to treat this patient population. In this cohort of NSQ NSCLC participants with high CEACAM5 expression on their tumors (≥50% with intensity ≥2), 10 of the 13 participants who responded had high baseline circulating CEA (≥100 ng / mL). In a cohort of NSQ NSCLC participants with moderate CEACAM5 expression on their tumors (≥1% and <50% of tumor cells with intensity ≥2+), potential response to tusamitamaravtansine was also evaluated, although only a limited number of participants (7 of 28 participants) had high circulating CEA levels at baseline.

[0682] CEACAM5 expression status may have differed between the initial tumor at diagnosis and the tumor at enrollment in the clinical trial. Although a correlation between circulating CEA and CEACAM5 expression status on tumors is expected, there are no available data on concomitant CEACAM5 expression on fresh tumor biopsies and circulating CEA. Biomarkers in oncology are most accurate and best correlated with clinical outcome when assessed immediately prior to treatment.

[0683] Participants are pre-screened into the second or third line EFC15858 trial based on CEACAM5 expression assessed on archival biopsy at diagnosis prior to first line treatment, which could be months or years prior to enrollment. This is potentially important for participants who are pre-screened into EFC15858 and have a negative diagnostic biopsy but may have high CEACAM5 expression upon fresh biopsy due to either heterogeneous expression of CEACAM5 or upregulation of CEACAM5 expression after first line treatment.

[0684] The study proposes fresh biopsy at screening (optional) to verify the correlation of CEACAM5 expression at diagnosis and enrollment, as well as the correlation of high circulating CEA and high CEACAM5 expression at enrollment. Fresh biopsies will be retrospectively evaluated due to tumor heterogeneity, and participants with negative CEACAM5 expression may have high expression in other tumor locations.

[0685] The aim of this study is to evaluate whether participants with high circulating CEA levels at baseline can benefit from tusamitama ravtansine treatment despite negative-moderate expression of CEACAM5 on archival biopsies, assuming that high circulating CEA levels reflect high CEACAM5 expression on tumors at enrollment.To this end, we will evaluate antitumor activity in participants with NSQ NSCLC who have high circulating CEA levels despite negative-moderate CEACAM5 expression on tumors by IHC testing in third or fourth line treatment.

[0686] Example 2. Evaluation of CEACAM5 expression on tumors using CEACAM5 IHC testing and circulating CEA and CEACAM5 assays Expression of CEACAM5 on tumors was assessed in patients enrolled in clinical trial TED13751 (NCT02187848) using CEACAM5 IHC testing and circulating CEA and CEACAM5 assays.

[0687] method A. Tumor CEACAM5 Immunohistochemistry (IHC) Testing: CEACAM5 tumor expression was assessed at pre-screening in the most recent available archived tumor samples (i.e., tumor tissue preserved at diagnosis, tumor tissue preserved at surgery, or tumor samples preserved prior to study inclusion and not undergoing anti-cancer treatment). The level and pattern of CEACAM5 expression in tumor tissue was determined using either IHC performed locally at the clinical site (prospective studies) and / or IHC performed centrally in the laboratory (retrospective or prospective studies). For local evaluation, the assay was performed using the anti-CEACAM5 clone 769 antibody provided by Sanofi. Anti-CEACAM5 clone 769 is a mouse monoclonal antibody with the same specificity as tusamitama ravtansine for the CEACAM5 target. For central evaluation, samples were first analyzed with a validated assay using the Sanofi anti-CEACAM5 clone 769 antibody running on the Techmate platform. In a second analysis, a validated CEACAM5 IHC assay was then performed on the Dako / Agilent Autostainer Link 48 IHC platform using the same anti-CEACAM5 antibody at the same concentration. For both assays, interpretation of CEACAM5 reactivity was performed by a certified pathologist using a semiquantitative percent score (calculated by summing the percentage of intensities ≧2+) or an H-score for CEACAM5 plasma membrane staining (total or polarized) in tumor cells. Cytoplasmic staining was also assessed.

[0688] B. Circulating CEA Assay: Circulating CEA was determined locally using an assay routinely used at the institution. The in vitro diagnostic CEA assay measures CEA and makes no claims of specificity for CEACAM5. Samples of circulating CEA were collected at baseline (during screening and any time prior to the first dose of tusamitamab ravtansine), then during treatment and at disease progression according to the same schedule as tumor assessments.

[0689] C. Circulating CEACAM5 Assay: Circulating CEACAM5 was determined centrally at the same time points before prescreening, at baseline (either within 7 days of dosing or at cycle 1, day 1 (C1D1) and at PK sampling during cycle 1 and before each dose of tusamitama ravtansine. Central testing specific for circulating CEACAM5 levels was performed at the sponsor's laboratory. The assay was a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) method using a rat monoclonal anti-human CEACAM5 antibody (clone GFR44) for capture and a sheep polyclonal anti-human CEACAM5 antibody conjugated to biotin for detection. In vitro evaluation of interference with tusamitama ravtansine at concentrations ranging from 0.100 ng / mL to 500 μg / mL showed no effect on quantification of CEACAM5 in human plasma. The quantification range was 120 pg / mL to 750,000,000 pg / mL (750 μg / mL).

[0690] result A. Circulating CEA / Circulating CEACAM5 Correlation: Circulating CEA and CEACAM5 levels at baseline for the high and intermediate NSQ NSCLC cohorts are shown in Figure 1. The correlation between baseline circulating CEA and circulating CEACAM5 levels was very strong (Spearman's rank correlation coefficient 0.99), indicating that the entity measured as circulating CEA in this patient population is closely related to circulating CEACAM5.

[0691] B. Baseline circulating CEA levels and responses: Overall response rates (ORR) according to circulating CEA levels are shown in Table 1. Forty percent of high expressers had high circulating CEA levels at baseline (>100 μg / L). Of the 13 responders, 10 (77%) had baseline circulating CEA levels >100 μg / L, indicating that high circulating CEA levels may predict response.

[0692] [Table 1]

[0693] C. CEACAM5 Tumor Expression / Circulating CEA Correlation: The correlation between circulating CEA levels at baseline and CEACAM5 expression by immunohistochemistry for the high and intermediate NSQ NSCLC cohorts is shown in Table 2.

[0694] [Table 2]

[0695] The correlation between circulating CEA and CEACAM5 expression in tumor tissues was weak in the two cohorts (Spearman's rank correlation coefficient was 0.402 in the high expression cohort and 0.343 in the medium expression cohort). This difference may be due to the difference in collection time points between the three samples (archival samples for IHC that may have been sufficiently sampled before the start of the study) and because CEACAM5 expression may be heterogeneous in tumors or may differ between primary tumors and metastases. Circulating CEA as a systemic assessment may reflect the total CEACAM5 derived from several lesions (e.g., primary or metastases).

[0696] Example 3. Study Design This is an open-label, non-randomized dose-escalation, safety, pharmacokinetic and antitumor activity evaluation of tusamitamaravutansine administered as a single agent by IV infusion, q2w or q3w (1 cycle = 3 weeks) in adults with advanced solid tumors. The escalation phase has three different cohorts: the main dose-escalation cohort receiving tusamitamaravutansine every 2 weeks, the escalation bis cohort receiving tusamitamaravutansine every 2 weeks with a loading dose in cycle 1 only, and the dose-escalation q3w cohort receiving tusamitamaravutansine every 3 weeks. The study is divided into three parts: the escalation phase and the expansion phase.

[0697] During the titration phase, the treated population is enriched for, but not limited to, patients with tumor types known to express CEACAM5, and confirmation of CEACAM5 expression is performed retrospectively using recent archived tissue samples and IHC at a central laboratory. Expression of circulating CEACAM5 can be used for enrichment.

[0698] The expansion phase will be performed on a twice weekly dosing schedule. During the expansion phase, the treated population will be restricted to patients with advanced disease of colorectal cancer (CRC), NSQ NSCLC, small cell lung cancer (SCLC) and gastric adenocarcinoma (including signet ring cell subtype and Siewert type II and III esophagogastric junction (EGJ) adenocarcinoma). CEACAM5 expression of intensity ≥2+ involving ≥50% of the tumor cell population will be demonstrated using local IHC evaluation of patients potentially eligible for study treatment during pre-screening on recent archived tissue samples and in the gastric adenocarcinoma cohort. There will be two independent NSQ NSCLC expansion phase cohorts. The first one (lung cohort) will include patients with high CEACAM5 expression (intensity ≥2+ involving at least 50% of the tumor cell population). The second cohort (lung bis cohort) will include patients pre-screened as positive with intermediate intensity (≥2+ involving ≥1% to <50% of the tumor cell population). The SCLC cohort will include patients with CEACAM5 expression of intensity ≥2+ comprising ≥1% of the tumor cell population. Pre-screening assessment of tumor CEACAM5 expression will be performed centrally for study centers that do not have a CEACAM5 assay - based on the tusamitama ravtansine monoclonal antibody - implemented in their local IHC platform. Full screening will be performed only for archived cases that meet the above definition. CEACAM5 pre-screening will not be required for CRC patients potentially eligible for the CRC expansion cohort. The level of CEACAM5 expression will be recorded centrally essentially retrospectively on both archived and fresh (baseline sample) tumor tissues. Confirmation of CEACAM5 tumor expression will be performed retrospectively in a central laboratory on fresh tumor tissue collected at baseline (mandatory biopsy for all patients with CRC and gastric cancer as well as expansion phase only for NSQ NSCLC and SCLC patients with lesions that can undergo biopsy). If sufficient archived tumor material is available, a central evaluation will also be performed retrospectively to gain knowledge of the variability of the evaluation of expression. The results of the retrospective analysis will not affect patient treatment.This will be used for better interpretation of overall response and as a baseline to compare with CEACAM5 expression at progression (exploring loss of CEACAM5 as a mechanism of acquired resistance).

[0699] With this goal, circulating CEACAM5 will be collected in all pre-screened NSCLC and SCLC patients, and data will also be collected in pre-screened failing patients (ie, CEACAM5 negative or expression <2+ intensity).

[0700] Approximately 3 mL of blood will be drawn for this sample and tested at a central laboratory during the pre-screening visit. Because this is a prospective descriptive diagnostic analysis, there is no statistical power for potential patients to be included, and all new pre-screened patients with NSQ NSCLC and SCLC will be candidates for this pre-screening CEACAM5 evaluation portion. Based on the potential target patients enrolled in the NSQ NSCLC and SCLC cohorts, approximately 562 patients (334 NSQ NSCLC patients and 228 SCLC patients) will be pre-screened.

[0701] Dose escalation decisions are based on dose-limiting toxicities (DLTs) observed during cycle 1 + cycle 2 of q2w cycles and cycle 1 of q3w cycles if not related to disease progression, whether related to the investigational medicinal product (IMP) or not, in the absence of clear evidence to the contrary, after review by the study committee. Before escalating the tusamitama ravtansine dose to the next level, safety data, especially DLTs, ocular and cardiac adverse events (AEs), will be reviewed by the study committee. Cumulative toxicities observed with subsequent doses will also be considered for the dose escalation process and dose selection decisions. The decision to dose escalate or deescalate will be based on the evaluation of DLTs, at least by consensus between the sponsor and the investigator.

[0702] The safety of a fixed dose of tusamitamaravtansine administered once q2w (on day 1 of each 14-day cycle) will be evaluated using a primary dose escalation process using accelerated dose escalation of the first three dose levels (DL), followed by Bayesian escalation with overdose control (EWOC).

[0703] For the accelerated dose escalation phase and for Bayesian EWOC, a minimum of 3 cycles (approximately 4 weeks) is required between the last patient receiving Cycle 1 Day 1 on DLn and the first patient receiving Cycle 1 Day 1 on DLn+1. The dose determined to be the maximum tolerated dose (MTD) will be further tested in expansion cohorts.

[0704] Loading dose of tusamitamab ravtansine on day 1 of cycle 1, followed by 100 mg / m 2 The safety of dosing with 120 mg / m2 (determined to be the MTD in the main escalation) will be evaluated in a dose escalation phase that will use a Bayesian EWOC process and will be run in parallel with the expansion phase. The first loading dose to be tested is 120 mg / m2. 2 If this initial dose is safe, dose escalation will proceed to the next 135 mg / m 2 and 150 mg / m 2 Proceed to the test of 150mg / m 2 If no DLT occurs at 170, 190, and 210 mg / m, the study committee may choose to continue with the study at 170, 190, and 210 mg / m if Bayes' rule allows it. 2 It may be decided to evaluate a higher DL of 210 mg / m 2 If no DLT occurs at this dose, the safety of this loading dose (MTD or maximum safe administered dose (MAD)) will be considered for the recommendation of the second phase dose at the end of the study. The determined MTD or safe MAD will be used for the CRC-L cohort.

[0705] The safety of tusamitama ravtansine administered once q3w (day 1 of each 21-day cycle) will be evaluated in a dose-escalation q3w process with a Bayesian EWOC design used as the primary dose escalation. The DLs tested in the model are 120, 150, 170, 190 and 210 mg / m 2The DL is 190 and 210 mg / m 2 are optional DLs, and escalation to these DLs will be determined by the Study Committee based on DLTs during Cycle 1 and cumulative safety and PK data and Bayesian modeling.

[0706] Inclusion criteria Locally advanced or metastatic solid malignant tumor disease for which, in the opinion of the investigator, standard alternative therapies are not available and which meets the following inclusion criteria:

[0707] At least 6x5μm slides from formalin-fixed paraffin-embedded (FFPE) archived tissue, and an additional number of slides that may be either 3x10μm (best) or 6x5μm, or equivalent to maintain the same total amount of material required, should be available for local testing at the site and / or shipment to the sponsor or a laboratory designated by the sponsor, for evaluation of tumor CEACAM5 expression (retrospectively in the expansion phase and prospectively in the expansion growth phase) and exploration of other response predictive biomarkers. In cases where less material is available, patients may still be eligible after discussion with the sponsor to evaluate and confirm that there is sufficient relevant material for the primary evaluation.

[0708] For participants in the accrual phase cohorts (main, bis, and q3w), enrollment is enriched for tumors that express or are likely to express CEACAM5, including (but not limited to): Malignancies with a high prevalence of CEACAM5 expression, i.e. CRC of adenocarcinoma or large cell subtype, NSQ NSCLC, gastric adenocarcinoma (including Siewert types II and III signet ring cell carcinoma and EGJ adenocarcinoma), or Malignant diseases with various degrees of prevalence and intensity of CEACAM5 expression include: cervical squamous cell carcinoma, pancreatic adenocarcinoma, bladder transitional cell carcinoma, cholangiocarcinoma, endometrial adenocarcinoma and epithelial ovarian carcinoma; · Circulating CEA levels >5ng / mL as demonstrated by local testing. For participants in the expansion phase cohorts, enrollment is limited to the following: Patients with either NSCLC of non-squamous subtype or gastric adenocarcinoma including signet ring cell carcinoma subtype and Siewert type II and III gastroesophageal junction (EGJ; equally gastroesophageal junction (GEJ)) adenocarcinoma. For the gastric adenocarcinoma cohort, CEACAM5 expression in the most recent FFPE archived tumor tissue sample is ≥2+ intensity in at least 50% of the tumor cell population as prospectively demonstrated by local IHC assessment. There are three independent cohorts in NSQ NSCLC expansion phase based on local or central CEACAM5 expression assessment on archived tumor tissue. The first one (lung) includes patients with CEACAM5 expression of ≥2+ intensity involving at least 50% of the tumor cell population. The second independent cohort (lung bis) includes patients with prescreen positive with ≥2+ intensity in ≥1% to <50% of the tumor cell population. · SCLC patients with CEACAM5 expression in the most recent FFPE archival tumor tissue sample with intensity ≥ 2+ involving ≥ 1% of the tumor cell population as prospectively demonstrated by local or central IHC evaluation. · Or all patients with CRC (including patients prescreened CEACAM5 positive regardless of expression level). Once the expansion phase is initiated, CRC patients must be preferentially included in the expansion CRC cohort unless they are ineligible (no target lesion or lesion suitable for biopsy) and must not be included in the dose escalation bis employed in parallel. At least one measurable disease by Response Evaluation Criteria in Solid Tumors version 1.1 (RECISTv1.1) in expansion phase only. At least one lesion suitable for biopsy (expansion cohort only - CRC and gastric cancer only). Patients must consent to a baseline biopsy for retrospective confirmation of tumor CEACAM5 expression before treatment initiation (except for NSCLC or no lesion suitable for biopsy).

[0709] Exclusion criteria Life expectancy <12 weeks. Known or symptomatic brain metastases (other than completely resected or previously irradiated non-progressive / recurrent) or leptomeningeal carcinomatosis. Concomitant treatment with any other anti-cancer therapy. Prior maytansinoid treatment (DM1 or DM4 antibody drug conjugates).

[0710] treatment Tusamitamavutansine is supplied as a concentrate with a 25 mL extractable volume for solution for injection of 125 mg (5 mg / mL) contained in a 30 mL Type I glass vial.

[0711] The initial starting DL is 5 mg / m administered q2w. 2 In the dose escalation q3w cohort, the starting DL is 120 mg / m 2 The patient's body surface area (BSA) is calculated using the patient's height and actual weight. BSA > 2.2m 2 For patients with 2 Calculated based on BSA.

[0712] Premedication with a histamine H1 antagonist (diphenylhydramine 50 mg PO or equivalent [e.g., dexchlorpheniramine] administered approximately 1 hour prior to administration of tusamitamab ravtansine) is required for all patients. Using an infusion-controlled pump, tusamitamab ravtansine is administered by IV infusion at a rate of 2.5 mg / min for the first 30 minutes, then increased to 5 mg / min in the absence of a hypersensitivity reaction.

[0713] Effectiveness Pharmacodynamic biomarker evaluation Tusamitama ravtansine is a cytotoxic agent with a selective delivery system that shows promise in inducing objective responses in the clinic. A single dose of tusamitama ravtansine induces regression in relevant experimental models.

[0714] However, the specificity associated with CEACAM5 targeting would potentially allow more accurate tracking of the impact of treatment on tumor growth by monitoring CEA levels in the blood. Indeed, measurement of circulating CEA levels is somewhat useful for CRC treatment monitoring. Usually, CEA returns to normal within 1-2 months after surgery, and if it does not return, it indicates disease recurrence. Similarly, modulation of circulating CEA may be a useful PDy marker of response to tusamitama ravtansine. An internal preclinical study evaluating circulating CEA in mice orthotopically implanted with patient-derived CRC xenografts expressing CEACAM5 showed a good correlation between levels of CEA and tumor burden, and treatment with a single active dose of tusamitama ravtansine prevented the rise in circulating CEA that occurs with tumor growth. The utility of monitoring circulating CEA will be explored in this clinical study.

[0715] Plasma samples of 2 mL or 3 mL will be collected at baseline and on treatment for all patients participating in the expansion cohort to determine the modulation of CEA levels under tusamitama ravtansine treatment. In cycle 1, sample collection will begin 6 hours after infusion to coincide with PK collection after tusamitama ravtansine. Samples for circulating CEA will be collected once before each subsequent dose, at the end of treatment, and at follow-up as needed. Determination of circulating CEACAM5 levels in PDy will be performed in a central laboratory designated by the sponsor using a commercially available ELISA kit. To assess intrapatient variability, one plasma sample will be collected within 7 days of enrollment (screening period), then pre-dose on day 1 of cycle 1.

[0716] Efficacy Evaluation Criteria In the escalation cohort, objective response information is obtained when patients have disease that can be easily measured and reassessed.

[0717] In the expansion cohort, the primary objective is to evaluate response to tusamitama ravtansine. All patients treated in the expansion phase must have at least one measurable lesion for enrollment. Tumor assessments will be performed at least every 4 cycles. The decision to continue treatment will be based on response assessment by the investigator, while lesion measurements will be collected in an electronic case report form for response determination by the sponsor. A partial or complete response must be confirmed with a second examination performed at least 4 weeks apart to be recorded as a confirmed response to treatment.

[0718] RECIST v1.1 criteria will be followed for evaluation of tumor response.

Claims

1. 1. An antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent for use in a method of treating cancer in a subject in need thereof, comprising: the anti-CEACAM5 antibody comprises an HCDR1 having the amino acid sequence of SEQ ID NO: 1, an HCDR2 having the amino acid sequence of SEQ ID NO: 2, an HCDR3 having the amino acid sequence of SEQ ID NO: 3, an LCDR1 having the amino acid sequence of SEQ ID NO: 4, an LCDR2 having the amino acid sequence NTR, and an LCDR3 having the amino acid sequence of SEQ ID NO: 5; The method includes measuring circulating carcinoembryonic antigen (CEA) in the subject, and if the circulating CEA in the subject is 5 ng / mL or greater, administering an effective amount of the ADC to the subject, thereby treating the cancer; the circulating CEA is measured after performing immunohistochemical analysis of CEACAM5 expression on tumor cells of the subject; The subject has negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in less than 1% of cells), or the subject has moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in more than 1% and less than 50% of cells), or the subject has high CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in more than 50% of cells).

2. 2. The antibody-drug conjugate (ADC) of claim 1, wherein the cytotoxic agent is selected from the group consisting of a taxoid, a vinca, a maytansinoid or maytansinoid analog, a small drug, a tomaymycin or pyrrolobenzodiazepine derivative, a cryptophycin derivative, a leptomycin derivative, an auristatin or dolastatin analog, a prodrug, a topoisomerase II inhibitor, a DNA alkylating agent, an antitubulin agent, and CC-1065 or a CC-1065 analog.

3. 2. The antibody-drug conjugate (ADC) of claim 1, wherein the cytotoxic agent is a maytansinoid or a maytansinoid analog.

4. 2. The antibody-drug conjugate (ADC) of claim 1, wherein the cytotoxic agent is N2'-deacetyl-N2'-(3-mercapto-1-oxopropyl)-maytansine (DM1) or N2'-deacetyl-N-2'(4-methyl-4-mercapto-1-oxopentyl)-maytansine (DM4).

5. The antibody-drug conjugate (ADC) of claim 1, wherein the anti-CEACAM5 antibody comprises a heavy chain variable domain (VH) having the amino acid sequence of SEQ ID NO: 6 and a light chain variable domain (VL) having the amino acid sequence of SEQ ID NO:

7.

6. 2. The antibody-drug conjugate (ADC) of claim 1, wherein the cancer is selected from the group consisting of colorectal cancer, gastric cancer, gastroesophageal junction cancer, esophageal cancer, lung cancer, cervical cancer, pancreatic cancer, ovarian cancer, thyroid cancer, bladder cancer, endometrial cancer, breast cancer, liver cancer, biliary tract cancer (e.g., cholangiocarcinoma), prostate cancer, and skin cancer.

7. 2. The antibody-drug conjugate (ADC) of claim 1, wherein the cancer is selected from the group consisting of colorectal cancer, gastric cancer, lung cancer, pancreatic cancer and prostate cancer.

8. The antibody-drug conjugate (ADC) of claim 1, wherein the cancer is lung cancer or gastric cancer.

9. before treatment, the circulating CEA is 20 ng / mL or greater; or before treatment, the circulating CEA is 50 ng / mL or greater; or before treatment, the circulating CEA is 80 ng / mL or greater; or 2. The antibody-drug conjugate (ADC) of claim 1, wherein prior to treatment, the circulating CEA is 100 ng / mL or greater.

10. The antibody-drug conjugate (ADC) of claim 1, wherein the anti-CEACAM5 antibody is tusamitamab.

11. The antibody-drug conjugate (ADC) of claim 1, which is tusamitamaravtansine.

12. The tusamitamaravtansine was administered at a dose of 100 mg / m 2 or the tusamitamavutansine is administered at a dose of 100 mg / m or more approximately once every two weeks. 2 The antibody-drug conjugate (ADC) of claim 11, administered at a dose of at least about once every three weeks.

13. 10. The antibody-drug conjugate (ADC) of claim 1, further comprising administering to the subject an effective amount of at least one additional agent effective in treating the cancer.

14. 14. The antibody-drug conjugate (ADC) of claim 13, wherein the additional agent is selected from the group consisting of immune checkpoint inhibitors (ICIs), platinum-based chemotherapy, pemetrexed, anti-VEGFR2, FOLFOX, FOLFIRI, TAS-102, anti-EGFR, cetuximab, and any combination thereof.

15. The antibody-drug conjugate (ADC) of claim 14, wherein the ICI is an anti-PD-1 antibody or an anti-PD-L1 antibody.

16. 16. The antibody-drug conjugate (ADC) of claim 15, wherein the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, sintilimab, dostallimab, and tislelizumab.

17. 16. The antibody-drug conjugate (ADC) of claim 15, wherein the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, and durvalumab.

18. The antibody-drug conjugate (ADC) of claim 1, comprising administering to the subject effective amounts of tusamitamaravtansine and pembrolizumab.

19. 20. The antibody-drug conjugate (ADC) of claim 18, further comprising administering to the subject an effective amount of platinum-based chemotherapy.

20. 20. The antibody-drug conjugate (ADC) of claim 19, wherein the platinum-based chemotherapy is selected from cisplatin and carboplatin.

21. 21. The antibody-drug conjugate (ADC) of claim 20, further comprising administering to the subject an effective amount of pemetrexed.

22. 1. An antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent for use in a method of treating cancer in a subject in need thereof, comprising: the anti-CEACAM5 antibody comprises an HCDR1 having the amino acid sequence of SEQ ID NO: 1, an HCDR2 having the amino acid sequence of SEQ ID NO: 2, an HCDR3 having the amino acid sequence of SEQ ID NO: 3, an LCDR1 having the amino acid sequence of SEQ ID NO: 4, an LCDR2 having the amino acid sequence NTR, and an LCDR3 having the amino acid sequence of SEQ ID NO: 5; the cytotoxic agent is a maytansinoid or a maytansinoid analog; The method includes measuring circulating carcinoembryonic antigen (CEA) in the subject, and if the circulating CEA in the subject is 5 ng / mL or greater, administering an effective amount of the ADC to the subject, thereby treating the cancer; the circulating CEA is measured after performing immunohistochemical analysis of CEACAM5 expression on tumor cells of the subject; The subject has negative or low CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in less than 1% of cells), or the subject has moderate CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in more than 1% and less than 50% of cells), or the subject has high CEACAM5 expression on tumor cells as measured by immunohistochemistry (intensity of ≧2+ in more than 50% of cells).

23. 1. An antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent for use in a method of treating cancer in a subject in need thereof, comprising: the ADC is tusamitamaravtansine; The method includes measuring circulating carcinoembryonic antigen (CEA) in the subject, and if the circulating CEA in the subject is 100 ng / mL or greater, administering to the subject an effective amount of the ADC, thereby treating the cancer; the circulating CEA is measured after performing immunohistochemical analysis of CEACAM5 expression on tumor cells of the subject; The subject has negative or low CEACAM5 expression on tumor cells (intensity of ≧2+ in less than 1% of cells) as measured by immunohistochemistry, or the subject has moderate CEACAM5 expression on tumor cells (intensity of ≧2+ in greater than 1% and less than 50% of cells) as measured by immunohistochemistry.

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