Use of anti-CEACAM5 immune complexes to treat neuroendocrine cancers expressing CEACAM5

Abstract

This disclosure provides a method for treating neuroendocrine cancer using an immune complex comprising hCEACAM5, and more particularly, an antibody that specifically binds to hCEACAM5.

Description

[Technical Field]

[0001] This disclosure relates to the field of therapeutic treatment of cancers such as neuroendocrine cancers that express CEACAM5, particularly hCEACAM5. Specific aspects of this disclosure relate to the use of CEACAM5 antagonists, such as anti-CEACAM5 antibodies and immune complexes, for the treatment of neuroendocrine cancers. [Background technology]

[0002] The mechanism of action of antibody-drug conjugates (ADCs) begins with their binding to specific antigens that are well-expressed on tumor cells, in order to achieve selective and efficient drug uptake. As demonstrated by the recent approvals of brentuximab vetotin for the treatment of Hodgkin lymphoma and trastuzumab emtansine (T-DM1) for the treatment of recurrent metastatic HER2+ breast cancer, selectively targeted ADCs with potent cytotoxicity against tumor cells have now proven to be an effective strategy for treating cancer. Many other malignancies, such as solid tumors, which do not fit the medical needs, could benefit from these treatment options.

[0003] Neuroendocrine carcinoma is a highly malignant disease defined as a neoplasm arising from poorly differentiated cells that express neuroendocrine markers (chromogranin, synaptophysin, INSM1, CD56, etc.) and exhibit high proliferative capacity (as evidenced by high mitotic index and Ki-67 index). Neuroendocrine neoplasms are classified according to the WHO classification based on differentiation and proliferation grade.

[0004] Currently, their management is based on standard first-line chemotherapy combining etoposide and cisplatin. After an initial response, all patients rapidly relapse, and second-line treatment is ineffective, with an average survival time of approximately 12–15 months.

[0005] It has been shown in the art that neuroendocrine cancers, such as prostate cancer and medullary thyroid carcinoma, express specific tumor markers (Lee et al., PNAS, March 28, 2018, 115(19)E4473-E4482 and Turkdogan et al., Journal of Otolaryngology-Head and Nech Surgery, 2018, 47:55). Lee et al. suggest that the cell surface proteins FXYD3 and CEACAM5 may be particularly useful targets for immuno-based therapy against advanced prostate cancer.

[0006] Carcinoembryonic antigens (CEAs) are glycoproteins involved in cell adhesion. CEAs are proteins normally expressed in the fetal intestines during the first six months of pregnancy and are found in pancreatic, liver, and colorectal cancers. The CEA family, consisting of 18 genes, is subdivided into two protein subgroups: the carcinoembryonic antigen-related cell adhesion molecule (CEACAM) subgroup and the pregnancy-specific glycoprotein subgroup (Kammerer & Zimmermann, BMC Biology 2010, 8:12).

[0007] In humans, the CEACAM subgroup consists of seven members: CEACAM1, CEACAM3, CEACAM4, CEACAM5, CEACAM6, CEACAM7, and CEACAM8. Numerous studies have shown that CEACAM5 is highly expressed on the surface of tumor cells in the colon, stomach, lung, breast, prostate, ovary, cervix, and bladder, and weakly expressed in a few normal epithelial tissues, such as columnar epithelium and goblet cells of the colon, mucous cervical cells of the stomach, and squamous epithelial cells of the esophagus and cervix (Hammarstroem et al, 2002, "Tumor markers, Physiology, Pathobiology, Technology and Clinical Applications," Eds. Diamandis EP et al., AACC Press, Washington, pp 375). Therefore, CEACAM5 may constitute a suitable therapeutic target for tumor-specific targeted approaches such as antibody-drug conjugates (ADCs).

[0008] Numerous anti-CEA antibodies have been developed for diagnostic or therapeutic purposes targeting CEA. Specificity to the relevant antigen has always been a point of interest in this field. International Publication No. 2014 / 079886 discloses an antibody that binds to the A3-B3 domains of human and cynomolgus monkey (Macaca fascicularis) CEACAM5 protein and does not significantly cross-react with human CEACAM1, human CEACAM6, human CEACAM7, human CEACAM8, cynomolgus monkey (Macaca fascicularis) CEACAM1, cynomolgus monkey (Macaca fascicularis) CEACAM6, and cynomolgus monkey (Macaca fascicularis) CEACAM8. This antibody binds to maytansinoid and thereby exhibits significant cytotoxic activity in MKN45 human gastric cancer cells. 50 The present invention provides antibody-drug conjugates (ADCs) (or immune complexes) with a value of 1 nM or less.

[0009] The use of anti-CEACAM-SN38 antibody-drug conjugates has been described in the Art and proposed as a therapeutic agent in the treatment of neuroendocrine prostate cancer (DeLucia et al., Clin Cancer Res, 2021, February 01, 27(3):759-774). These conjugates have a specific structure and mechanism of action. SN-38 is derived from camptothecin and is an irinotecan analog, a DNA topoisomerase type I inhibitor. [Overview of the Initiative] [Problems that the invention aims to solve]

[0010] There is a continuing need for the development of novel therapies and more effective strategies to treat CEACAM5-expressing neuroendocrine cancers. [Means for solving the problem]

[0011] This disclosure provides, in particular, a method for treating neuroendocrine cancers of subjects in need, comprising administering an effective amount of an antibody or immune complex (including an antibody) that specifically binds to CEACAM5.

[0012] In some embodiments, the Disclosure relates to an immune complex comprising an antibody or an antigen-binding fragment thereof for use in treating cancers selected from neuroendocrine cancers expressing hCEACAM5 in subjects where such treatment is necessary, wherein the antibody or the antigen-binding fragment is specifically bound to hCEACAM5, and the antibody or the antigen-binding fragment is bound to or linked to at least one growth inhibitor, the growth inhibitor being not a topoisomerase I inhibitor.

[0013] In some embodiments, the antibody or its antigen-binding fragment comprises a VH domain and a VL domain, the VH domain comprising three complementarity-determining regions (CDRs) HCDR1, HCDR2, and HCDR3, and the VL domain comprising three CDRs LCDR1, LCDR2, and LCDR3. HCDR1 is the amino acid sequence of SEQ ID NO: 3 [ka] HCDR2 contains the amino acid sequence of SEQ ID NO: 4 [ka] HCDR3 contains the amino acid sequence of SEQ ID NO: 5 [ka] Includes, LCDR1 is the amino acid sequence of SEQ ID NO: 6 [ka] LCDR2 contains the amino acid sequence of NTR, and LCDR3 contains the amino acid sequence of SEQ ID NO: 7 [ka] Includes.

[0014] In some embodiments, the growth inhibitor is selected from the group consisting of chemotherapeutic agents, enzymes, antibiotics and small molecule toxins or enzymatically active toxins, taxoids, vinca, taxanes, maytansinoids or maytansinoid analogs, tomaimycin or pyrrolobenzodiazepine derivatives, cryptophycin derivatives, leptomycin derivatives, auristatin or drastatin analogs, prodrugs, topoisomerase II inhibitors, DNA alkylating agents, antitubulin agents and toxins such as CC-1065 or CC-1065 analogs.

[0015] In some embodiments, the Disclosure relates to an immune complex comprising an antibody or an antigen-binding fragment thereof for use in treating cancers selected from neuroendocrine cancers expressing hCEACAM5 in subjects where such treatment is necessary, wherein the antibody or the antigen-binding fragment thereof specifically binds to hCEACAM5, and the antibody or the antigen-binding fragment thereof comprises a VH domain and a VL domain, the VH domain comprising three complementarity-determining regions (CDRs) HCDR1, HCDR2 and HCDR3, and the VL domain comprising three CDRs LCDR1, LCDR2 and LCDR3. HCDR1 is the amino acid sequence of SEQ ID NO: 3 [ka] HCDR2 contains the amino acid sequence of SEQ ID NO: 4 [ka] HCDR3 contains the amino acid sequence of SEQ ID NO: 5 [ka] Includes, LCDR1 is the amino acid sequence of SEQ ID NO: 6 [ka] LCDR2 contains the amino acid sequence of NTR, and LCDR3 contains the amino acid sequence of SEQ ID NO: 7 [ka] Includes, This neuroendocrine cancer is not small cell lung carcinoma (SCLC).

[0016] In some embodiments, neuroendocrine cancers are selected from the group consisting of neuroendocrine cancers of the esophagus, stomach, small and large intestine, anal region, pancreas, bladder, female reproductive organs, male reproductive organs, thyroid gland, and head and neck.

[0017] In another aspect, the Disclosure relates to an antibody, or an antigen-binding fragment thereof, or an immune complex comprising such antibody or antigen-binding fragment, for use in treating cancers selected from neuroendocrine cancers expressing hCEACAM5 in subjects where such treatment is necessary, wherein the antibody specifically binds to hCEACAM5, and the antibody or antigen-binding fragment comprises a VH domain and a VL domain, the VH domain comprising three complementarity-determining regions (CDRs) HCDR1, HCDR2, and HCDR3, and the VL domain comprising three CDRs LCDR1, LCDR2, and LCDR3. HCDR1 contains the amino acid sequence of SEQ ID NO: 3 (GFVFSSYD), HCDR2 contains the amino acid sequence of SEQ ID NO: 4 (ISSGGGIT), and HCDR3 contains the amino acid sequence of SEQ ID NO: 5 (AAHYFGSSGPFAY). LCDR1 contains the amino acid sequence of SEQ ID NO: 6 (ENIFSY), LCDR2 contains the amino acid sequence of NTR, and LCDR3 contains the amino acid sequence of SEQ ID NO: 7 (QHHYGTPFT).

[0018] In certain embodiments, cancer is selected from neuroendocrine tumors (NETs), neuroendocrine carcinomas (NECs), mixed neuroendocrine-non-neuroendocrine neoplasms (MiNENs), pheochromocytomas, and medullary thyroid carcinomas (MTCs).

[0019] In particular, cancer - Neuroendocrine cancers of the gastrointestinal tract, particularly the esophagus, stomach, pancreas, liver, small intestine, and large intestine or anal region, or neuroendocrine cancers of the urinary tract, particularly neuroendocrine cancers of the bladder, and neuroendocrine cancers of the gastrointestinal tract and pancreatic and biliary ducts. - Neuroendocrine cancers of the upper respiratory tract, gastrointestinal tract, and salivary glands, including neuroendocrine cancers of the lung, particularly small cell lung cancer, and neuroendocrine cancers of the thymus. -Medullary thyroid carcinoma (MTC) and other neuroendocrine cancers of the thyroid gland, - Neuroendocrine cancers of the adrenal gland, such as pheochromocytoma; - Neuroendocrine cancers of the skin, such as Merkel cell carcinoma (MCC), - Neuroendocrine cancer of the female reproductive organs such as the endometrium, cervix, or ovaries. - Neuroendocrine cancers of the prostate or testes, especially the testes and other male reproductive organs, and - Neuroendocrine cancer of the head and neck Selected from.

[0020] In particular, cancers are selected from neuroendocrine cancers of the esophagus, stomach, small and large intestine, anal region, pancreas, bladder, lung, female reproductive organs, male reproductive organs, and head and neck. More specifically, cancers are neuroendocrine cancers of the lung, in particular neuroendocrine small cell lung cancer, or neuroendocrine cancers of the male reproductive organs, in particular neuroendocrine prostate cancer.

[0021] In some embodiments, the cancer is neuroendocrine small cell lung cancer. In other embodiments, the cancer is neuroendocrine prostate cancer.

[0022] In some embodiments, the cancer is not neuroendocrine small cell lung cancer. In other embodiments, the cancer is not neuroendocrine prostate cancer.

[0023] In certain embodiments, subjects may be moderate or high expressionrs of carcinoembryonic antigen-associated cell adhesion molecules. In particular, subjects may be moderate CEACAM5 expressionrs, i.e., subjects having CEACAM5 expression of 2+ or 3+ intensity as measured by hCEACAM5 immunohistochemistry in at least 1% of the tumor cell population, or subjects having CEACAM5 expression of 1+ intensity in at least 50% of the tumor cell population. In particular, subjects may be high CEACAM5 expressionrs, i.e., subjects having CEACAM5 expression of 2+ or 3+ intensity as measured by hCEACAM5 immunohistochemistry in more than 50% of the tumor cell population.

[0024] In a particular embodiment, the VH domain is: Sequence ID 1: [ka] Includes.

[0025] Antibodies or immune complexes are, SEQ ID NO: 8 [ka] It may contain heavy chains.

[0026] In a particular embodiment, the VL domain is: Sequence ID 2: DIQMTQSPASLSASVGDRVTITCRASENIFSYLAWYQQKPGKSPKLLVYNTRTLAEGVPSRFSGSGSGTDFSLTISSLQPEDFATYYCQHHYGTPFTFGSGTKLEIK Includes.

[0027] Antibodies or immune complexes are identified by SEQ ID NO: 9: [ka] It may include a light chain containing [something].

[0028] In certain embodiments, the antibody in an antibody-containing immune complex is conjugated or linked to at least one growth inhibitor. In particular, the growth inhibitor may be a cytotoxic agent. In particular, the growth inhibitor may be selected from the group consisting of chemotherapeutic agents, enzymes, antibiotics and small molecule toxins or enzymatically active toxins, taxoids, vinca, taxanes, maytansinoids or maytansinoid analogs, tomaimycin or pyrrolobenzodiazepine derivatives, cryptophycin derivatives, leptomycin derivatives, auristatin or drastatin analogs, prodrugs, topoisomerase inhibitors, DNA alkylating agents, antitubulin agents and toxins such as CC-1065 or CC-1065 analogs. In some embodiments, the topoisomerase inhibitor is not a topoisomerase I inhibitor. In some embodiments, the topoisomerase inhibitor is a topoisomerase II inhibitor.

[0029] In certain embodiments, the growth inhibitor is (N 2 ’-deacetyl-N 2 ’-(3-mercapto-1-oxopropyl)-maytansine) (DM1) or N 2 ’-deacetyl-N 2 ’-(4-methyl-4-mercapto-1-oxopentyl)-maytansine (DM4).

[0030] In certain embodiments, the antibody is covalently bound to at least one growth inhibitor via a cleavable or non-cleavable linker.

[0031] In certain embodiments, the immune complex comprises a heavy chain consisting of SEQ ID NO: 8 and a light chain consisting of SEQ ID NO: 9, and is covalently bound to an hCEACAM5 antibody conjugated to N 2 ’-deacetyl-N 2 ’(4-methyl-4-mercapto-1-oxopentyl)-maytansine (DM4) via succinimidyl pyridyldithiobutyrate (SPDB).

[0032] In certain embodiments, the immune complex is tusamitamab ravtansine.

[0033] In certain embodiments, the antibody or immune complex is administered at a dose level of 5, 10, 20, 30, 40, 60, 80, 100, 120, 150, 180 or 210 mg / m2 based on the body surface area of the subject. In certain embodiments, the antibody or immune complex is administered every two weeks or every three weeks. In certain embodiments, the antibody or immune complex is administered at a dose level of 100 mg / m2 every two weeks based on the body surface area of the subject. In certain embodiments, the antibody or immune complex is administered as a loading dose of about 100 mg / m 2 ~ about 200 mg / m 2 , particularly as a loading dose, about 135 mg / m 2 , about 150 mg / m 2 or a dose of 170 mg / m 2 .

[0034] This specification further describes a method for treating cancers selected from neuroendocrine cancers that express CEACAM5 in subjects where such treatment is necessary, the method comprising administering an antibody or its antigen-binding fragment, or an immune complex comprising the antibody or said antigen-binding fragment, wherein the antibody specifically binds to hCEACAM5, the antibody comprises a VH domain and a VL domain, the VH domain comprises three complementarity-determining regions (CDRs) HCDR1, HCDR2 and HCDR3, and the VL domain comprises three CDRs LCDR1, LCDR2 and LCDR3. HCDR1 contains the amino acid sequence of SEQ ID NO: 3 (GFVFSSYD), HCDR2 contains the amino acid sequence of SEQ ID NO: 4 (ISSGGGIT), and HCDR3 contains the amino acid sequence of SEQ ID NO: 5 (AAHYFGSSGPFAY). LCDR1 contains the amino acid sequence of SEQ ID NO: 6 (ENIFSY), LCDR2 contains the amino acid sequence of NTR, and LCDR3 contains the amino acid sequence of SEQ ID NO: 7 (QHHYGTPFT).

[0035] In particular, cancer may be selected from neuroendocrine neoplasms (NETs), neuroendocrine carcinomas (NECs), mixed neuroendocrine-nonneuroendocrine neoplasms (MiNENs), medullary thyroid carcinoma (MTC), and pheochromocytoma. In particular, cancer may be selected from - Neuroendocrine cancers of the gastrointestinal tract and pancreatic and biliary ducts, such as neuroendocrine cancers of the digestive tract, especially the esophagus, stomach, pancreas, liver, small intestine, and large intestine or anal region, or neuroendocrine cancers of the urinary tract, especially neuroendocrine cancers of the bladder. - Neuroendocrine cancers of the upper respiratory tract, gastrointestinal tract, and salivary glands, including neuroendocrine cancers of the lung, particularly small cell lung cancer, and neuroendocrine cancers of the thymus. -Medullary thyroid carcinoma (MTC) and other neuroendocrine cancers of the thyroid gland, - Neuroendocrine cancers of the adrenal gland, such as pheochromocytoma, - Neuroendocrine cancers of the skin, such as Merkel cell carcinoma (MCC), - Neuroendocrine cancer of the female reproductive organs such as the endometrium, cervix, or ovaries. - Neuroendocrine cancers of the male reproductive organs such as the prostate or testes, and - Neuroendocrine cancer of the head and neck Selected from.

[0036] In particular, the cancer may be selected from neuroendocrine cancers of the esophagus, stomach, small and large intestine, anal region, pancreas, bladder, lung, female reproductive organs, male reproductive organs, and head and neck. In particular, the cancer may be neuroendocrine cancer of the lung, especially neuroendocrine small cell lung cancer, or neuroendocrine cancer of the male reproductive organs, especially neuroendocrine prostate cancer. In some embodiments, the cancer is neuroendocrine small cell lung cancer. In other embodiments, the cancer is neuroendocrine prostate cancer.

[0037] In some embodiments, the cancer is not neuroendocrine small cell lung cancer. In other embodiments, the cancer is not neuroendocrine prostate cancer.

[0038] In certain embodiments, subjects are moderate or high expressionrs of carcinoembryonic antigen-associated cell adhesion molecules. In particular, subjects may be moderate CEACAM5 expressors, i.e., subjects having CEACAM5 expression of 2+ or 3+ intensity as measured by CEACAM5 immunohistochemistry in at least 1% of the tumor cell population, or subjects having CEACAM5 expression of 1+ intensity in at least 50% of the tumor cell population. In particular, subjects may be high CEACAM5 expressors, i.e., subjects having CEACAM5 expression of 2+ or 3+ intensity as measured by CEACAM5 immunohistochemistry in more than 50% of the tumor cell population.

[0039] In certain embodiments, the antibody is conjugated or linked to at least one growth inhibitor. In particular, the growth inhibitor is (N 2 '-deacetyl-N 2 '-(3-mercapto-1-oxopropyl)-meitansine)(DM1) or N 2 '-deacetyl-N 2 It may also be -(4-methyl-4-mercapto-1-oxopentyl)-meytansine (DM4).

[0040] In certain embodiments, the antibody comprises a heavy chain consisting of SEQ ID NO: 8 and a light chain consisting of SEQ ID NO: 9, and is covalently bound to N2'-deacetyl-N2'(4-methyl-4-mercapto-1-oxopentyl)-meytansine (DM4) via N-succinimidylpyridiyl dithiobutyrate (SPDB) and is an hCEACAM5 antibody. In particular, the antibody may be tusamitamabrabutansine.

[0041] In certain embodiments, the antibody or immune complex is administered at dose levels of 5, 10, 20, 30, 40, 60, 80, 100, 120, 150, 180, or 210 mg / m2 based on the target body surface area. In certain embodiments, the antibody or immune complex is administered every two weeks or every three weeks. In certain embodiments, the antibody or immune complex is administered at a dose level of 100 mg / m2 every two weeks based on the target body surface area. In certain embodiments, the antibody or immune complex is administered at a loading dose of approximately 100 mg / m2. 2 ~about 200mg / m 2 In particular, the loading dose is approximately 135 mg / m². 2 , about 150mg / m 2 , or 170 mg / m² 2 It is administered in the following dosage. [Brief explanation of the drawing]

[0042] [Figure 1] This graph shows CEACAM5 expression measured by RNA in PDx models. The horizontal axis represents each PDx model, and the vertical axis represents the expression level of CEACAM5 with respect to FPKM (fragments per kilobase transcript per over 1 million mapped fragments). [Figure 2]The graph shows the change in tumor volume (mm3) over time (days) (n=7 or 8) after administration of unrelated DM4-ADC immune complexes at a concentration of 35 mg / kg in different mouse models (rounded square), or after administration of tusamitamabrabutansine (also referred to as SAR408701 in the figure) at a concentration of 5 mg / kg (rounded upright triangle). A) Model MR-0009-RO (NE+ / CEACAM+ phenotype), B) Model MR-0084 (NE+ / localized CEACAM5 phenotype), C) Model MR-0123 (ADK+ / CEACAM- phenotype), D) Model MR-0191 (NE+ / CEACAM+ phenotype), and E) Model MR-059 (NE+ / CEACAM5+ phenotype). *, **, ***, and *** indicate significance of the results. [Figure 2-1] Same as above. [Figure 2-2] Same as above. [Figure 3] The graphs show the change in body weight (g) over time (days) (n=7 or 8) in different mouse models: a control model without any administration (rounded square), a different mouse model after administration of an unrelated DM4-ADC immune complex at a concentration of 35 mg / kg (rounded upright triangle), or a different mouse model after administration of tusamitamabrabutansine (also referred to as SAR408701 in the figure) at a concentration of 5 mg / kg (rounded inverted triangle). A) Model MR-0009-RO, B) Model MR-0084, C) Model MR-0123, D) Model MR-0191, and E) Model MR-059. [Figure 3-1] Same as above. [Figure 3-2] Same as above. [Figure 4A]Figures 4(A and B) show the correlation between CEACAM5 expression (measured by FPKM from RNA sequencing) and NEPC markers (measured by FPKM from RNA sequencing), for example, AR (Pearson r=-0.4214, p=0.0001), SYP (Pearson r=0.2617, p=0.0085), KLK3 (Pearson r=-0.4976, p=0.0001), ASCL1 (Pearson r=0.6352, p=0.0001), FOLH1 (Pearson r=-0.2584, p=0.0094), and DLL3 (Pearson r=0.5486, p=0.0001). [Figure 4B] Figures 4(A and B) show the correlation between CEACAM5 expression (measured by FPKM from RNA sequencing) and NEPC markers (measured by FPKM from RNA sequencing), for example, AR (Pearson r=-0.4214, p=0.0001), SYP (Pearson r=0.2617, p=0.0085), KLK3 (Pearson r=-0.4976, p=0.0001), ASCL1 (Pearson r=0.6352, p=0.0001), FOLH1 (Pearson r=-0.2584, p=0.0094), and DLL3 (Pearson r=0.5486, p=0.0001). [Modes for carrying out the invention]

[0043] This disclosure provides pharmaceutical compositions and methods for using these compositions for the treatment of neuroendocrine cancers expressing CEACAM5, as well as improvement of at least one symptom of the disease. These compositions comprise at least one antibody that specifically binds to CEACAM5, for example, the antibody huMAb2-3. ADC huMAb2-3-SPDB-DM4 is an immune complex combining the huMAb2-3 (anti-CEACAM5) antibody with a meitansinoid derivative 4 (DM4), a potent antimitotic agent that inhibits microtubule aggregation. DM4 is stable in plasma and covalently binds to huMAb2-3 via an optimized linker SPDB [N-succinimidyl 4-(2-pyridyldithio)-butyrate] that is cleavable in cells. After binding and uptake in targeted cancer cells, huMAb2-3-SPDB-DM4 (tusamitamabrabutansine) is degraded, releasing cytotoxic DM4 metabolites.

[0044] The inventors of this invention have found that many neuroendocrine cancers express the cell surface protein CEACAM5.

[0045] As can be seen in the experimental data further provided herein, proof of concept was achieved in PDx models of neuroendocrine prostate cancer treated with tusamitamablubutansine, showing a significant reduction in tumor volume in all models and no significant toxicity observed with the treatment. In particular, CEACAM5 expression was shown to correlate with the expression of neuroendocrine prostate cancer markers.

[0046] Further proof of concept was achieved by the inventors in a subset of neuroendocrine small cell lung cancers treated with tusamitamablubutansine. In particular, these data support the conclusion that tusamitamablubutansine is effective in treating neuroendocrine SCLC.

[0047] These data support the conclusion that tusamitamabrabutansine is particularly effective in treating high-CEACAM5-expressing neuroendocrine cancers.

[0048] definition Where used herein, the term “about” in quantitative terms refers to ±10% of the value it modifies (rounded up to the nearest integer if the value is not divisible, for example, the number of molecules or nucleotides). For example, the phrase “about 100 mg” would encompass 90 mg to 110 mg (both ends). The phrase “about 2500 mg” would encompass 2250 mg to 2750 mg. Where applied to percentages, the term “about” refers to plus or minus 10% of that percentage. For example, the phrase “about 20%” would encompass 18 to 22%, and “about 80%” would comprehensively encompass 72 to 88% (both ends). Furthermore, where “about” is used herein in conjunction with quantitative terms, it is understood that, in addition to the plus or minus 10% value, the exact value of the quantitative term is also intended and included. For example, the term “about 23%” explicitly intends, describes, and includes exactly 23%.

[0049] It should be noted that the term “one (a)” or “one (an)” entity refers to one or more of those entities. For example, “antibody” is understood to represent one or more antibodies. Therefore, the terms “one (a)” (or “one (an)”), “one or more” and “at least one” may be used interchangeably herein.

[0050] Throughout this specification and its embodiments, variations of the terms “have” and “comprise,” or “has,” “having,” “comprises,” or “comprises,” will be understood to mean that they encompass the specified integer or group of integers, but do not exclude any other integer or group of integers. Variations of the terms “have” and “comprise,” or “has,” “having,” “comprises,” or “comprises,” will be understood to mean that they encompass one or more specified elements (such as a composition of a substance or a method step), but do not exclude any other elements. The term “consists of” means that it encompasses one or more specified elements, but excludes any additional elements. The term “essentially consists of” means that it encompasses the specified elements and, where applicable, other elements that do not substantially affect the fundamental characteristics of this disclosure. Various embodiments of this disclosure that use the term "includes" or its equivalent are understood to include embodiments in which this term is replaced with "includes only," "consists of," or "essentially consists of."

[0051] Whenever an aspect is described herein with the term "comprising," it should be understood that similar aspects described with the terms "consisting of" and / or "essentially consisting of" are also provided.

[0052] Furthermore, as used herein, “and / or” should be interpreted as the specific disclosure of each of two particular features or components that have or do not have the other. Accordingly, as used herein, the term “and / or” in phrases such as “A and / or B” is intended to include “A and B,” “A or B,” “A” (alone) and “B” (alone). Similarly, when the term “and / or” is used in phrases such as “A, B and / or C,” it is intended to include 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 (alone); B (alone); and C (alone).

[0053] Whenever an aspect is described herein with the word "comprising," it should be understood that similar aspects described with the terms "consisting of" and / or "essentially consisting of" are also provided.

[0054] An antibody can be a natural or conventional antibody, consisting of two heavy chains linked to each other by disulfide bonds, with each heavy chain linked to a light chain by disulfide bonds. There are two types of light chains: lambda(l) and kappa(k). There are five major heavy chain classes (or isotypes) that determine the functional activity of antibody molecules: IgM, IgD, IgG, IgA, and IgE. Each chain contains different sequence domains. The light chain contains two domains or regions: a variable domain (VL) and a constant domain (CL). The heavy chain contains four domains, a variable domain (VH), and three constant domains (CH1, CH2, and CH3, collectively called CH). The variable regions of both the light chain (VL) and the heavy chain (VH) determine the binding recognition and specificity to the antigen. The constant domains of the light chain (CL) and heavy chain (CH) confer important biological properties such as antibody chain association, secretion, transplacental migration, complement binding, and binding to the Fc receptor (FcR). The Fv fragment is the N-terminal portion of the Fab fragment of immunoglobulins and consists of a variable region of one light chain and one heavy chain. Antibody specificity lies in the structural complementarity between the antibody binding site and the antigenic determinant. The antibody binding site is mainly composed of residues derived from the hypervariable or complementarity-determining region (CDR). Occasionally, residues derived from the non-hypervariable region or framework region (FR) affect the entire domain structure and, therefore, the binding site. Thus, the complementarity-determining region or CDR refers to the amino acid sequence that together defines the binding affinity and specificity of the native Fv region of the native immunoglobulin binding site. The light and heavy chains of immunoglobulins each contain three CDRs, called CDR1-L, CDR2-L, CDR3-L, and CDR1-H, CDR2-H, and CDR3-H, respectively. Therefore, a conventional antibody-antigen binding site contains six CDRs, including sets of CDRs from the heavy chain V region and the light chain V region, respectively.

[0055] The "framework region" (FR) refers to the amino acid sequence interposed between CDRs, i.e., the relatively conserved portions of the immunoglobulin light chain variable region and immunoglobulin heavy chain variable region across different immunoglobulins of the same species. The light and heavy chains of immunoglobulins each have four FRs, called FR1-L, FR2-L, FR3-L, FR4-L, and FR1-H, FR2-H, FR3-H, and FR4-H, respectively. Human framework regions are substantially identical (approximately 85% or more, particularly 90%, 95%, 97%, 99%, or 100%) to the framework regions of naturally occurring human antibodies.

[0056] In the context of this disclosure, the definition of CDR / FR in immunoglobulin light chains or heavy chains should be determined based on the definition by IMGT (Lefranc et al. Dev.Comp.Immunol., 2003, 27(1):55-77; www.imgt.org).

[0057] As used herein, the term “antibody” refers to conventional antibodies and their fragments, as well as single-domain antibodies and their fragments, in particular variable heavy chains of single-domain antibodies, and chimeric, humanized, bispecific, or multispecific antibodies.

[0058] As used herein, antibody or immunoglobulin also includes “single-domain antibodies,” which are antibodies whose complementarity-determining region is part of a single-domain polypeptide, as described more recently. Examples of single-domain antibodies include heavy-chain antibodies, antibodies that naturally lack a light chain, single-domain antibodies derived from conventional four-chain antibodies, and engineered single-domain antibodies. Single-domain antibodies may be derived from any species, including but not limited to mice, humans, camels, llamas, goats, rabbits, and cattle. Single-domain antibodies may be naturally occurring single-domain antibodies known as light-chain-lacking heavy-chain antibodies. In particular, camelid species, such as camels, dromedaries, llamas, alpacas, and guanacos, produce light-chain-lacking heavy-chain antibodies. Camel heavy-chain antibodies also lack the CH1 domain.

[0059] The variable heavy chains of these single-domain antibodies lacking light chains are known in the art as "VHH" or "nanobody(registered trademark)". Similar to conventional VH domains, VHH contains four FRs and three CDRs. VHH has advantages over conventional antibodies. They are about 10 times smaller than IgG molecules, and as a result, well-folded functional VHH can be produced by in vitro expression with high yield. Furthermore, VHH is very stable and resistant to protease action. The characterization and production of VHH were carried out as described by Harmsen and De Haard HJ (Appl. Microbiol. Biotechnol. 2007 Nov;77(1):13-22).

[0060] As used herein, the terms “monoclonal antibody” or “mAb” refer to an antibody molecule with a single amino acid sequence against a specific antigen and should not be interpreted as requiring antibody production by any particular method. Monoclonal antibodies can be produced by a single clone of a B cell or hybridoma, but they can also be produced by recombinants, i.e., protein engineering.

[0061] The term "humanized antibody" refers to an antibody that is entirely or partially of non-human origin and has been modified to evade or minimize the immune response in humans, particularly by substituting specific amino acids in the framework regions of the VH and VL domains. The constant domains of humanized antibodies are, in most cases, the human CH and CL domains.

[0062] A (conventional) antibody "fragment" is a part of an intact antibody, particularly the antigen-binding region or variable region of the intact antibody. Examples of antibody fragments include Fv, Fab, F(ab')2, Fab', dsFv, (dsFv)2, scFv, sc(Fv)2, diabodies, bispecific and multispecific antibodies formed from antibody fragments. Conventional antibody fragments can also be heavy chain antibodies or single-domain antibodies such as VHH.

[0063] The term "Fab" refers to an antibody fragment with a molecular weight of approximately 50,000 and antigen-binding activity, in which approximately half of the N-terminal side of the heavy chain and the entire light chain are linked by disulfide bonds. It is typically obtained by treating IgG with a protease such as papain.

[0064] The term "F(ab')2" refers to an antibody fragment with a molecular weight of approximately 100,000 and slightly greater antigen-binding activity than two identical Fab fragments linked via a disulfide bond in the hinge region. It is typically obtained by treating IgG with a protease such as pepsin.

[0065] The term "Fab'" refers to an antibody fragment with a molecular weight of approximately 50,000 and antigen-binding activity, obtained by cleaving the disulfide bond in the hinge region of F(ab')2.

[0066] A single-stranded Fv ("scFv") polypeptide is a VH::VL heterodimer typically expressed from a gene fusion containing VH and VL encoding genes linked by a covalently bonded peptide-encoding linker. The human scFv fragments of this disclosure contain a CDR preserved in an appropriate conformation, particularly by using recombination techniques. Divalent and multivalent antibody fragments can be spontaneously formed by the association of monovalent scFv or produced by coupling monovalent scFv with a peptide linker such as divalent sc(Fv)2. "dsFv" is a VH::VL heterodimer stabilized by a disulfide bond. "(dsFv)2" indicates two dsFv coupled by a peptide linker.

[0067] The term "bispecific antibody" or "BsAb" refers to an antibody that combines the antigen-binding sites of two antibodies within a single molecule. Therefore, a BsAb can bind to two different antigens simultaneously. Genetic engineering is increasingly used to design, modify, and produce antibodies or antibody derivatives with a desired set of binding properties and effector functions, as described, for example, in European Patent Application Publication No. 2050764A1.

[0068] The term "multispecific antibody" refers to an antibody that combines two or more antibody antigen-binding sites within a single molecule.

[0069] The term "diabody" refers to a small antibody fragment having two antigen-binding sites, which contains a heavy chain variable domain (VH) linked to a light chain variable domain (VL) in the same polypeptide chain (VH-VL). By using a linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with a complementary domain on another chain, creating two antigen-binding sites.

[0070] An amino acid sequence that is "at least 85% identical to the reference sequence" is a sequence that, in its entire length, has sequence identity of 85% or more, particularly 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% with the full-length reference amino acid sequence.

[0071] The percentage of "sequence identity" between amino acid sequences can be determined by comparing two sequences that are optimally aligned across a comparison window. The portion of the polynucleotide or polypeptide sequence within the comparison window may contain additions or deletions (i.e., gaps) compared to a reference sequence (which does not contain additions or deletions) for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions where identical nucleic acid bases or amino acid residues exist in both sequences, obtaining the number of matched positions, dividing the number of matched positions by the total number of positions within the comparison window, and multiplying the result by 100 to obtain the percentage of sequence identity. Optimal alignment of sequences for comparison is performed by global pairwise alignment, for example, using the algorithm in Needleman and Wunsch J.Mol.Biol.48:443 (1970). The percentage of sequence identity can be easily determined, for example, using the program Needle with the BLOSUM62 matrix and the following parameters: gap-open=10, gap-extend=0.5.

[0072] A "conservative amino acid substitution" is a substitution in which an amino acid residue is replaced by another amino acid residue having a side chain R group with similar chemical properties (e.g., charge, size, or hydrophobicity). Generally, conservative amino acid substitutions do not substantially alter the functional properties of a protein. Examples of amino acids with side chains having similar chemical properties include: 1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; 2) aliphatic-hydroxyl side chains: serine and threonine; 3) amide-containing side chains: asparagine and glutamine; 4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; 5) basic side chains: lysine, arginine, and histidine; 6) acidic side chains: aspartic acid and glutamic acid; and 7) sulfur-containing side chains: cysteine ​​and methionine. Conservative amino acid substitutions can also be defined based on amino acid size.

[0073] "Purified" and "isolated," when referring to polypeptides (i.e., the antibodies of this disclosure) or nucleotide sequences, mean that the indicated molecule exists in the substantial absence of other biological macromolecules of the same type. As used herein, the term "purified" specifically means the presence of at least 75%, 85%, 95%, or 98% (by weight) of the same type of biological macromolecule. An "isolated" nucleic acid molecule encoding a particular polypeptide refers to a nucleic acid molecule that substantially contains no other nucleic acid molecules that do not encode the polypeptide of interest. However, the molecule may contain several additional bases or parts that do not adversely affect the fundamental characteristics of the composition.

[0074] As used herein, the term “Subject” means mammals such as rodents, cats, dogs, and primates. In particular, the subject as disclosed herein is humans.

[0075] Antibody-drug conjugate containing anti-CEACAM5 antibody This disclosure relates to an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody for the treatment of cancers selected from neuroendocrine cancers that express hCEACAM5 in subjects where such treatment is necessary.

[0076] An antibody-drug conjugate typically comprises an anti-CEACAM5 antibody and at least one chemotherapeutic agent, such as a cell proliferation inhibitor. The antibody-drug conjugate (ADC) contains an anti-CEACAM5 antibody conjugated to at least one chemotherapeutic agent. In particular, in an antibody-drug conjugate, the anti-CEACAM5 antibody is covalently bound to at least one chemotherapeutic agent via a cleavable or non-cleavable linker.

[0077] Anti-CEACAM5 antibody This disclosure relates to the use of an antibody or its antigen-binding fragment that specifically conjugates CEACAM5 for treating neuroendocrine cancers expressing CEACAM5. In particular, the antibody or its antigen-binding fragment conjugates hCEACAM5.

[0078] According to a first aspect, the Disclosure relates to an antibody or an antigen-binding fragment thereof, or an immune complex comprising an antibody or said antigen-binding fragment, for use in treating cancers selected from neuroendocrine cancers expressing hCEACAM5 in subjects where such treatment is necessary, wherein the antibody specifically binds to CEACAM5, particularly hCEACAM5, and the antibody comprises a VH domain and a VL domain, the VH domain comprising three complementarity-determining regions (CDRs) HCDR1, HCDR2, and HCDR3, and the VL domain comprising three CDRs LCDR1, LCDR2, and LCDR3. HCDR1 contains the amino acid sequence of SEQ ID NO: 3 (GFVFSSYD), HCDR2 contains the amino acid sequence of SEQ ID NO: 4 (ISSGGGIT), and HCDR3 contains the amino acid sequence of SEQ ID NO: 5 (AAHYFGSSGPFAY). LCDR1 contains the amino acid sequence of SEQ ID NO: 6 (ENIFSY), LCDR2 contains the amino acid sequence of NTR, and LCDR3 contains the amino acid sequence of SEQ ID NO: 7 (QHHYGTPFT).

[0079] According to one embodiment, the antibody is a humanized anti-CEACAM5 antibody.

[0080] According to this disclosure, the anti-CEACAM5 antibody comprises a VH domain and a VL domain, the VH domain comprises three complementarity-determining regions (CDRs) HCDR1, HCDR2, and HCDR3, and the VL domain comprises three CDRs LCDR1, LCDR2, and LCDR3. HCDR1 is the amino acid sequence of SEQ ID NO: 3 [ka] HCDR2 contains the amino acid sequence of SEQ ID NO: 4 [ka] HCDR3 contains the amino acid sequence of SEQ ID NO: 5 [ka] Includes, LCDR1 is the amino acid sequence of SEQ ID NO: 6 [ka] LCDR2 contains the amino acid sequence of NTR, and LCDR3 contains the amino acid sequence of SEQ ID NO: 7 [ka] Includes.

[0081] In certain embodiments, the VH domain includes Sequence ID No. 1 (EVQLQESGPGLVKPGGSLSLSCAASGFVFSSYDMSWVRQTPERGLEWVAYISSGGGITYAPSTVKGRFTVSRDNAKNTLYLQMNSLTSEDTAVYYCAAHYFGSSGPFAYWGQGTLVTVSS).

[0082] In certain embodiments, the VL domain includes Sequence ID No. 2 (DIQMTQSPASLSASVGDRVTITCRASENIFSYLAWYQQKPGKSPKLLVYNTRTLAEGVPSRFSGSGSGTDFSLTISSLQPEDFATYYCQHHYGTPFTFGSGTKLEIK).

[0083] In further embodiments, the anti-CEACAM5 antibody includes a variable domain of the heavy chain (VH) consisting of SEQ ID NO: 1.

[0084] In further embodiments, the anti-CEACAM5 antibody includes a variable domain of the light chain (VL) consisting of SEQ ID NO: 2.

[0085] In particular, anti-CEACAM5 antibodies, -array [ka] A variable domain of the heavy chain consisting of, -array It contains a variable light chain domain consisting of DIQMTQSPASLSASVGDRVTITCRASENIFSYLAWYQQKPGKSPKLLVYNTRTLAEGVPSFSGSGSGTDFSLTISSLQPEDFATYYCQHHYGTPFTFGSGTKLEIK (SEQ ID NO: 2).

[0086] In a further embodiment, the anti-CEACAM5 antibody comprises a variable heavy chain (VH) domain having at least 90% identity with SEQ ID NO: 1, and a variable light chain (VL) domain having at least 90% identity with SEQ ID NO: 2, wherein HCDR1 is derived from SEQ ID NO: 3, HCDR2 from SEQ ID NO: 4, HCDR3 from SEQ ID NO: 5, LCDR1 from SEQ ID NO: 6, LCDR2 from the amino acid sequence NTR, and LCDR3 from SEQ ID NO: 7.

[0087] In a further embodiment, the anti-CEACAM5 antibody comprises a variable heavy chain (VH) domain having at least 92%, at least 95%, and at least 98% identity with SEQ ID NO: 1, and a variable light chain (VL) domain having at least 92%, at least 95%, and at least 98% identity with SEQ ID NO: 2, wherein HCDR1 consists of SEQ ID NO: 3, HCDR2 consists of SEQ ID NO: 4, HCDR3 consists of SEQ ID NO: 5, LCDR1 consists of SEQ ID NO: 6, LCDR2 consists of the amino acid sequence NTR, and LCDR3 consists of SEQ ID NO: 7.

[0088] In certain embodiments, the anti-CEACAM5 antibody includes a heavy chain (HC) containing SEQ ID NO: 8.

[0089] In further embodiments, the anti-CEACAM5 antibody comprises a light chain (LC) containing SEQ ID NO: 9.

[0090] In a further embodiment, the anti-CEACAM5 antibody includes a heavy chain (HC) consisting of SEQ ID NO: 8.

[0091] In a further embodiment, the anti-CEACAM5 antibody includes a light chain (LC) consisting of SEQ ID NO: 9.

[0092] In particular, anti-CEACAM5 antibodies, -array [ka] A heavy chain consisting of, -array [ka] It includes a light chain consisting of [a specific component].

[0093] In a further embodiment, the anti-CEACAM5 antibody comprises a heavy chain (HC) having at least 90% sequence identity with SEQ ID NO: 8, and a light chain (LC) having at least 90% sequence identity with SEQ ID NO: 9, wherein HCDR1 consists of SEQ ID NO: 3, HCDR2 consists of SEQ ID NO: 4, HCDR3 consists of SEQ ID NO: 5, LCDR1 consists of SEQ ID NO: 6, LCDR2 consists of the amino acid sequence NTR, and LCDR3 consists of SEQ ID NO: 7.

[0094] In a further embodiment, the anti-CEACAM5 antibody comprises a heavy chain (HC) having at least 92%, at least 95%, and at least 98% sequence identity with SEQ ID NO: 8, and a light chain (LC) having at least 92%, at least 95%, and at least 98% sequence identity with SEQ ID NO: 9, wherein HCDR1 consists of SEQ ID NO: 3, HCDR2 consists of SEQ ID NO: 4, HCDR3 consists of SEQ ID NO: 5, LCDR1 consists of SEQ ID NO: 6, LCDR2 consists of the amino acid sequence NTR, and LCDR3 consists of SEQ ID NO: 7.

[0095] The anti-CEACAM5 antibody may be a single-domain antibody or a fragment thereof. In particular, the single-domain antibody fragment may consist of a variable heavy chain (VHH) containing HCDR1, HCDR2, and HCDR3 of the above antibody. The antibody may also be a heavy-chain antibody, i.e., an antibody lacking a light chain, which may or may not contain the CH1 domain.

[0096] A single-domain antibody or fragment thereof may include the framework region of a camelid single-domain antibody and, optionally, the constant domain of a camelid single-domain antibody.

[0097] Anti-CEACAM5 antibodies may be antibody fragments selected from the group consisting of Fv, Fab, F(ab')2, Fab', dsFv, (dsFv)2, scFv, sc(Fv)2, and diabodies, and may particularly be humanized antibody fragments.

[0098] The antibody may also be a bispecific or multispecific antibody formed from antibody fragments, where at least one antibody fragment is an antibody fragment according to the present disclosure. A multispecific antibody is a multivalent protein complex, such as those described, for example, in European Patent Application Publication No. 2050764A1 or U.S. Patent Application Publication No. 2005 / 0003403A1.

[0099] Anti-CEACAM5 antibodies and fragments thereof can be produced by any technique known in the art. In particular, such antibodies can be produced by techniques as described below.

[0100] Anti-CEACAM5 antibodies and their fragments can be isolated (e.g., purified) from or contained within a vector, such as a membrane or lipid vesicle (e.g., liposome).

[0101] Anti-CEACAM5 antibodies and fragments thereof can be produced alone or in combination by any technique known in the art, for example, any chemical, biological, genetic, or enzymatic technique, though not limited to these.

[0102] By knowing the amino acid sequence of a desired sequence, those skilled in the art can readily produce anti-CEACAM5 antibodies and their fragments using standard techniques for polypeptide production. For example, they can be synthesized using well-known solid-phase methods, particularly using commercially available peptide synthesizers (e.g., those manufactured by Applied Biosystems, Foster City, California), according to the manufacturer's instructions. Alternatively, anti-CEACAM5 antibodies and their fragments can be synthesized by recombinant DNA techniques, as is well known in the art. For example, these fragments can be obtained as DNA expression products after incorporating the DNA sequence encoding the desired (poly)peptide into an expression vector and introducing such a vector into a suitable eukaryote or prokaryotic host expressing the desired polypeptide, from which they can later be isolated using well-known techniques.

[0103] Anti-CEACAM5 antibodies and their fragments can be appropriately isolated from the culture medium by conventional immunoglobulin purification procedures such as protein A-Sepharose chromatography, hydroxyapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.

[0104] Methods for producing humanized antibodies based on conventional recombinant DNA and gene transfection techniques are well known in the art (see, for example, Riechmann L. et al. 1988; Neuberger MS. et al. 1985). Antibodies can be humanized using various techniques known in the art, including techniques disclosed in applications such as International Publication No. 2009 / 032661, CDR grafting (European Patent No. 239,400, PCR Publication No. WO91 / 09967, U.S. Patents No. 5,225,539, No. 5,530,101 and No. 5,585,089), lamination or re-attachment (European Patent No. 592,106, European Patent No. 519,596, Padlan EA (1991); Studnicka GM et al. (1994); Roguska MA. et al. (1994)), and chain shuffling (U.S. Patent No. 5,565,332). Common recombinant DNA techniques for preparing such antibodies are also known (see European Patent Application No. 125023 and International Publication No. 96 / 02576).

[0105] The anti-CEACAM5 antibody Fab can be obtained by treating an antibody that specifically reacts with CEACAM5 with a protease such as papain. Alternatively, the anti-CEACAM5 antibody Fab can be produced by inserting the DNA sequences encoding both strands of the anti-CEACAM5 antibody Fab into a prokaryotic or eukaryotic expression vector, and then introducing the vector into prokaryotic or eukaryotic cells (if necessary) to express the anti-CEACAM5 antibody Fab.

[0106] Anti-CEACAM5 antibody F(ab')2 can be obtained by treating an antibody that specifically reacts with CEACAM5 with a protease and pepsin. Alternatively, anti-CEACAM5 antibody F(ab')2 can be produced by conjugating it to the following Fab' via a thioether bond or disulfide bond.

[0107] The Fab' of an anti-CEACAM5 antibody can be obtained by treating F(ab')2, which specifically reacts with CEACAM5, with a reducing agent such as dithiothreitol. Alternatively, the Fab' of an anti-CEACAM5 antibody can be produced by inserting the DNA sequence encoding the Fab' chain of the antibody into a prokaryotic or eukaryotic expression vector, and then introducing this vector into prokaryotic or eukaryotic cells (if necessary) for expression.

[0108] The scFv of an anti-CEACAM5 antibody can be produced by obtaining the sequences of the CDR or VH and VL domains as described above, constructing DNA encoding the scFv fragment, inserting the DNA into a prokaryotic or eukaryotic expression vector, and then introducing the expression vector into prokaryotic or eukaryotic cells (if necessary) to express the scFv. A well-known technique called CDR grafting can be used to produce humanized scFv fragments, which includes selecting the complementarity-determining regions (CDRs) according to this disclosure and grafting them onto a human scFv fragment framework of a known three-dimensional structure (e.g., International Publication No. 98 / 45322; International Publication No. 87 / 02671; U.S. Patent No. 5,859,205; U.S. Patent No. 5,585,089; U.S. Patent No. 4,816,567; European Patent No. 0173494).

[0109] In one embodiment, the anti-CEACAM5 antibody is tusamitamab (CAS[2349294-95-5]).

[0110] Chemotherapy agents This disclosure also includes cytotoxic complexes, or immune complexes, or antibody-drug complexes, or complexes. As used herein, these terms all have the same meaning and are interchangeable.

[0111] The antibody-drug conjugates for use as provided in this disclosure typically comprise at least one chemotherapeutic agent. As used herein, chemotherapeutic agents refer to agents that kill cells, including cancer cells. Such agents preferably halt the division and growth of cancer cells and reduce the size of tumors. The term "chemotherapeutic agent" is used herein interchangeably with the terms "cytotoxic agent," "proliferation inhibitor," or "cell growth inhibitor" or "cell suppressor."

[0112] Accordingly, this disclosure relates to an “immune complex” comprising an antibody of this disclosure conjugated or linked to at least one growth inhibitor. In certain embodiments, the growth inhibitor is a cytotoxic agent or a radioisotope.

[0113] The terms "proliferation inhibitor" or "antiproliferation agent" can be used interchangeably and refer to compounds or compositions that inhibit the growth of cells, particularly tumor cells, either in vitro or in vivo.

[0114] In further embodiments, the chemotherapeutic agent is selected from the group consisting of radioisotopes, protein toxins, small molecule toxins, and combinations thereof.

[0115] Examples of radioactive isotopes include those suitable for treating cancer. Such radioactive isotopes generally emit primarily beta radiation. In further embodiments, the radioactive isotope is 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 radioactive isotope is selected from the group consisting of radioactive isotopes of Lu and combinations thereof. In one embodiment, the radioactive isotope is an α radioactive isotope, more specifically Th 227 This emits alpha radiation.

[0116] The immune complexes described herein can be prepared as described in International Publication No. 2004 / 091668.

[0117] In further embodiments, the small molecule toxin is selected from antimetabolites, DNA alkylating agents, DNA crosslinking agents, DNA intercalating agents, microtubule inhibitors, topoisomerase inhibitors, and combinations thereof.

[0118] In further embodiments, the microtubule inhibitor is selected from the group consisting of taxanes, vinca alkaloids, maytansinoids, colchicine, podophyllotoxin, gluceofulvin, and combinations thereof.

[0119] As used herein, the terms “cytotoxic agent” or “growth inhibitor” refer to substances that inhibit or prevent the function of cells and / or cause cell destruction. 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.

[0120] In certain embodiments, the “cytotoxic agent” or “proliferation inhibitor” is selected from the group consisting of chemotherapeutic agents, enzymes, antibiotics and small molecule toxins or enzymatically active toxins, taxoids, vinca, taxanes, maytansinoids or maytansinoid analogs, tomaimycin or pyrrolobenzodiazepine derivatives, cryptophycin derivatives, leptomycin derivatives, auristatin or drastatin analogs, prodrugs, topoisomerase inhibitors, DNA alkylating agents, antitubulin agents and toxins such as CC-1065 or CC-1065 analogs. In some embodiments, the topoisomerase inhibitor is not a topoisomerase I inhibitor. In some embodiments, the topoisomerase inhibitor is a topoisomerase II inhibitor.

[0121] In certain embodiments, the growth inhibitor is not a topoisomerase inhibitor.

[0122] In certain embodiments, the growth inhibitor is selected from the group consisting of chemotherapeutic agents, enzymes, antibiotics and small molecule toxins or enzymatically active toxins, taxoids, vinca, taxanes, maytansinoids or maytansinoid analogs, tomaimycin or pyrrolobenzodiazepine derivatives, cryptophycin derivatives, leptomycin derivatives, auristatin or drastatin analogs, prodrugs, DNA alkylating agents, antitubulin agents and toxins such as CC-1065 or CC-1065 analogs.

[0123] An analog as understood herein is a compound that may be chemically different from another compound but shares the same or similar functional properties, in particular the same or similar biological properties as the compound in question.

[0124] As used herein, “enzymatically active toxins” are a type of toxin produced by certain organisms, such as bacteria, fungi, plants, or animals, that function by interfering with essential biological processes within the cells or tissues of the targeted organism, similar to how enzymes act. The enzymatic activity of these toxins is often a key mechanism by which they harm the host organism. Examples of enzymatically active toxins include botulinum toxin, diphtheria toxin, cholera toxin, Shiga toxin, and tetanus toxin.

[0125] As used herein, “taxoid” or “taxane” refers to two classes of taxol-derived derivatives developed for their anti-cancer chemotherapy properties. Examples of such compounds include, but are not limited to, paclitaxel, docetaxel, cabazitaxel, paclitaxel prigmex, paclitaxel docohexaenoic acid, paclitaxel trevatide, AI-850, mirataxel, and ANG1005.

[0126] As used herein, "vinca" or "vinca alkaloids" refers to a set of anti-mitotic and anti-microtubule alkaloids originally derived from the Vinca plant that block β-tubulin polymerization in dividing cells, and have therefore been used in cancer chemotherapy. Examples of vinca include, but are not limited to, vinblastine, vincristine, vindesine, vinorelbine, vincaminol, vineridine, and vinbrunin.

[0127] Pyrrolobenzodiazepines (PBDs) are naturally occurring anticancer drug molecules consisting of a central pyrrolo[2,1-c][1,4]benzodiazepine core. Examples of PBDs and PBD derivatives include, but are not limited to, tomaimycin, diazepam, anthramycin, cibilomycin, and polotoramycin.

[0128] Cryptophycins are a family of macrolide molecules that are potent cytotoxins and are being studied for their potential antiproliferative properties that could be useful in developing chemotherapy. Examples of cryptophycins and their derivatives include, but are not limited to, cryptophycin-1, cryptophycin-52 (LY355703), and cryptophycin-55.

[0129] As used herein, "leptomycin derivative" refers to a member of the leptomycin family as defined by Kalese et al. in Synthesis 2002, 8, 981-1003, including leptomycin A and leptomycin B, callistatin, latijadone such as latijadone A and latijadone B, anginomycin such as anginomycin A, B, C, and D, causamycin, leptolstatin, and leptofranins such as leptofranin A, B, C, and D.

[0130] As used herein, "drastatin" and "aulistin" refer to the linear and cyclic peptides first isolated from D. auricularia. Among these, certain drastatins have shown effective cytotoxic effects on tumor cells. Examples of drastatin derivatives include, but are not limited to, drastatin-10, drastatin-15, drastatinol, auristatin, symprostatin 1, and symprostatin 5.

[0131] As used herein, “prodrug” is a pharmacologically inactive drug or compound that is metabolized to a pharmacologically active drug after ingestion. Type I prodrugs are physiologically activated intracellularly, while type II prodrugs are physiologically activated extracellularly, particularly in digestive fluids or in the body’s circulatory system, particularly in the blood. Examples of type I prodrugs include, but are not limited to, acyclovir, fluorouracil, cyclophosphamide, diethylstilbustrol diphosphate, L-DOPA, mercaptopurine, mitomycin, zidovudine, carbamazepine, captopril, carisoprodol, heroin, morcidomin, leflunomide, paliperidone, phenacetin, primidone, psilocybin, sulindac, and fursultiamine. Examples of type II prodrugs include, but are not limited to, loperamide oxide, oxyphenisatin, sulfasalazine, acetylsalicylate, bacampicillin, bambuterol, chloramphenicol succinate, dipivefrin, fosphenytoin, lisdexamfetamine, pralidoxime, ADEPT, GDEPT, and VDEPT.

[0132] Topoisomerases are enzymes that have evolved to resolve topological problems in DNA by transiently disrupting one or both strands of DNA. Topoisomerase I catalyzes changes in DNA topology via transient single-strand breaks, while topoisomerase II catalyzes changes in DNA topology via transient double-strand breaks. Both human topoisomerase I and topoisomerase II (both α and β isoforms) can be targeted in anticancer chemotherapy using topoisomerase inhibitors.

[0133] Examples of topoisomerase I as used herein include, but are not limited to, camptothecin (CPT) and its analogues such as deluxtecan, topotecan, irinotecan, SN-38, siratecan, cocitecan, exatecan, lulutotecan, gimatecan, berothecan, and rubitecan.

[0134] Examples of topoisomerase II as used herein include, but are not limited to, anthracyclines, etoposide, teniposide, doxorubicin, daunorubicin, epirubicin, idarubicin, mitoxantrone, ICRF-193, ICRF-187 (dexrazoxane), melbaron, and acralubicin.

[0135] As used herein, “DNA alkylating agent” refers to a class of antitumor or anticancer agents that act by inhibiting the transcription of DNA into RNA, thereby halting protein synthesis. Examples of such compounds include, but are not limited to, altretamine, bendamustine, busulfan, carmustine, chlorambucil, cyclophosphamide, dacarbazine, ifosfamide, lomustine, rubinectedine, mechloretamine, melphalan, procarbazine, streptozosin, temozolomide, thiotepa, trabectedine, and platinum-coordinated conjugates such as carboplatin, cisplatin, and oxaliplatin.

[0136] Antitubulin agents constitute a large class of compounds that exhibit broad activity in both solid tumors and hematological malignancies due to their interference with microtubule dynamics. Examples of such compounds include, but are not limited to, taxanes, vinca alkaloids, drastatin, estramustine, meitansinoids, halichondrin, nocodazole, cryptophycin, colchicine and its analogs, hemiasterin, podophyllotoxin, combretastatin, 2-methoxyestradiol, 4-substituted methoxybenzoyl-arylthiazole (SMAT), phenylrahistine, steganacin, and clacin.

[0137] CC-1065 is a naturally occurring antitumor agent with a unique structure produced by Streptomyces zelensis. CC-1065 analogs are a class of alkylating agents that are highly suitable for targeted tumor therapy. Examples of CC-1065 analogs include, but are not limited to, adzeresin, calceresin, and bis-indole-(seco)-CBI3 derivatives.

[0138] In some embodiments, the cytotoxic agent is a taxoid, vinca, maytansinoid or maytansinoid analog such as DM1 or DM4, small drugs, tomaimycin or pyrrolobenzodiazepine derivatives, cryptophycin derivatives, leptomycin derivatives, auristatin or drastatin analogs, prodrugs, topoisomerase inhibitors, particularly topoisomerase II inhibitors, DNA alkylating agents, antitubulin agents, CC-1065 or CC-1065 analogs.

[0139] In some embodiments, the cytotoxic agent may be a meitansinoid.

[0140] As used herein, "maytansinoid" means maytansinoid and maytansinoid analogs. Maytansinoids are drugs that inhibit microtubule formation and are highly toxic to mammalian cells.

[0141] Suitable examples of maytansinoids include maytansinol, maytansinol analogs, and combinations thereof.

[0142] Suitable examples of meitansinol analogs include those having a modified aromatic ring and those having modifications at other positions. Such suitable meitansinoids are listed 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; This information is disclosed in Specification No. 4,362,663; Specification No. 4,364,866; Specification No. 4,450,254; Specification No. 4,322,348; Specification No. 4,371,533; Specification No. 6,333,410; Specification No. 5,475,092; Specification No. 5,585,499; and Specification No. 5,846,545.

[0143] Specific examples of suitable analogs of meitansinol having a modified aromatic ring include: (1) C-19-dechloro(U.S. Patent No. 4,256,746) (prepared by LAH reduction of anthamitocin P2); (2) C-20-hydroxy(or C-20-demethyl)+ / -C-19-dechloro(U.S. Patent No. 4,361,650 and No. 4,307,016) (prepared by demethylation using Streptomyces or Actinomyces or dechlorination using LAH); and (3) C-20-demethoxy, C-20-acyloxy (-OCOR), + / -dechloro (U.S. Patent No. 4,294,757) (prepared by acylation using acyl chloride).

[0144] Specific examples of suitable analogues of meitansinol with other positional modifications include: (1) C-9-SH (U.S. Patent No. 4,424,219) (prepared by reaction of meitansinol with H2S or P2S5); (2) C-14-alkoxymethyl (demethoxy / CH2OR) (U.S. Patent No. 4,331,598); (3) C-14-hydroxymethyl or acyloxymethyl (CH2OH or CH2Oac) (U.S. Patent No. 4,450,254) (prepared from Nocardia); (4) C-15-hydroxy / acyloxy (U.S. Patent No. 4,364,866) (prepared by conversion of meitansinol by Streptomyces); (5) C-15-methoxy (US Patent No. 4,313,946 and No. 4,315,929) (isolated from Trewia nudiflora); (6) C-18-N-demethyl (U.S. Patent Nos. 4,362,663 and 4,322,348) (prepared by demethylation of meitansinol by Streptomyces); and (7) 4,5-deoxy (U.S. Patent No. 4,371,533) (prepared by LAH reduction of titanium trichloride / maytancinol).

[0145] In one embodiment of the present disclosure, the cytotoxic complex of the present disclosure is formally N as a cytotoxic agent. 2’ -deacetyl-N 2’ The thiol-containing maytansinoid (DM1) called -(3-mercapto-1-oxopropyl)-maytansine is used. DM1 is represented by the following structural formula (I). [ka]

[0146] In another embodiment, the cytotoxic complex of the present disclosure is formally N as the cytotoxic agent. 2’ -deacetyl-N- 2’The thiol-containing maytansinoid DM4, called (4-methyl-4-mercapto-1-oxopentyl)-maytansine, is used. DM4 is represented by the following structural formula (II). [ka]

[0147] In certain embodiments, the inhibitor or growth inhibitor is (N2'-deacetyl-N2'-(3-mercapto-1-oxopropyl)-meytansine) (DM1) or N2'-deacetyl-N2'-(4-methyl-4-mercapto-1-oxopentyl)-meytansine (DM4).

[0148] In further embodiments of the present disclosure, other maytansins can be used, including thiols and disulfide-containing maytansinoids having mono- or dialkyl substitutions on the carbon atoms having sulfur atoms. These include maytansinoids having acylated amino acid side chains having acyl groups having hindered sulfhydryl groups at C-3, C-14 hydroxymethyl, C-15 hydroxy, or C-20 desmethyl, wherein the carbon atoms of the acyl groups having thiol functional groups have one or two substituents, such substituents are linear or branched alkyl or alkenyl groups having CH3, C2H5, 1 to 10 reagents, and any aggregates that may be present in solution.

[0149] Examples of these cytotoxic agents and methods of binding are further shown in International Publication No. 2008 / 010101, which is incorporated by reference.

[0150] Therefore, in further embodiments, the maytansinoid is selected from the group consisting of (N2'-deacetyl-N2'-(3-mercapto-1-oxopropyl)-maytansine)DM1 or N2'-deacetyl-N-2'(4-methyl-4-mercapto-1-oxopentyl)-maytansine (DM4) and combinations thereof.

[0151] In some embodiments, the antibodies of the Disclosure are covalently bound to at least one growth inhibitor or cytotoxic agent, either directly or via a cleavable or non-cleavable linker.

[0152] In particular, the antibody is covalently bound to at least one growth inhibitor via a linker that is either cleavable or incleavable.

[0153] As used herein, "linker" means a chemical moiety that contains a chain of covalent bonds or atoms that covalently bond a polypeptide to the drug moiety.

[0154] The conjugate may be prepared by an in vitro method. A binding group is used to bind the drug or prodrug to the antibody. Suitable binding groups are well known in the art and include disulfide groups, thioether groups, acid-unstable groups, photo-unstable groups, peptidase-unstable groups, and esterase-unstable groups. The binding of the antibody to the cytotoxic agent or growth inhibitor of this disclosure is not limited to, but includes, N-succinimidylpyridiyl dithiobutyrate (SPDB), 4-[(5-nitro-2-pyridinyl)dithio]-2,5-dioxo-1-pyrrolidinyl ester (nitro-SPDB), 4-(pyridine-2-yldisulfanyl)-2-sulfobutyrate (sulfo-SPDB), N-succinimidyl (2-pyridyldithio)propionate (SPDP), succinimidyl (N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), iminothiolane (IT), and imide. These may be prepared using various bifunctional protein coupling agents, including difunctional derivatives of esters (such as dimethyladipimidate HCl), active esters (such as diserate subtinimidyl), aldehydes (such as glutaraldehyde), bis-azide compounds (such as bis(p-azidobenzoyl)-hexanediamine), bis-diazonium derivatives (such as bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (e.g., toluene 2,6-diisocyanate), and bis-active fluorine compounds (e.g., 1,5-difluoro-2,4-dinitrobenzene). For example, lysine immunotoxins can be prepared as described in Vitetta et al (1987). Carbon-labeled 1-isothiocyanatobenzylmethyldiethylenetriaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for the conjugation of radioactive nucleotides to antibodies (International Publication No. 94 / 11026).

[0155] The linker may be a “cleavable linker” that facilitates the release of chemotherapeutic agents in cells. For example, acid-unstable linkers, peptidase-sensitive linkers, esterase-unstable linkers, photosensitive linkers, or disulfide-containing linkers (see, for example, U.S. Patent No. 5,208,020) can be used. The linker may also be a “non-cleavable linker” (e.g., an SMCC linker) which may provide better resistance in some cases.

[0156] In certain embodiments, the linker is selected from the group consisting of N-succinimidylpyridiyl dithiobutyrate (SPDB), 4-(pyridine-2-yldisulfanyl)-2-sulfobutyrate (sulfo-SPDB), and succinimidyl (N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC).

[0157] In further embodiments, the linker binds to a lysine or cysteine ​​residue in the Fc region of the anti-CEACAM5 antibody. In even further embodiments, the linker forms a disulfide bond or thioether bond with maytansine.

[0158] The linker may be a “cleavable linker” that facilitates the release of cytotoxic agents or growth inhibitors within cells. For example, acid-unstable linkers, peptidase-sensitive linkers, esterase-unstable linkers, photosensitive linkers, or disulfide-containing linkers (see, for example, U.S. Patent No. 5,208,020) can be used. The linker may also be a “non-cleavable linker” (e.g., an SMCC linker) which may provide better resistance.

[0159] Alternatively, the fusion protein comprising the antibody and the cytotoxic or growth-inhibiting polypeptide of this disclosure may be prepared by recombinant technology or peptide synthesis. The length of the DNA may include each region encoding two parts of the complex that are adjacent to each other or separated by regions encoding linker peptides that do not disrupt the desired properties of the complex.

[0160] The antibodies of this disclosure may also be used in enzyme-dependent prodrug therapy by conjugating polypeptides to prodrug-activating enzymes that convert prodrugs (e.g., peptidyl chemotherapeutic agents, see International Publication No. 81 / 01145) into active anticancer drugs (e.g., see International Publication No. 88 / 07378 and U.S. Patent No. 4,975,278). The enzymatic components of immune complexes useful in ADEPT include any enzymes that can act on the prodrug to convert it into its more active cytotoxic form. Enzymes useful in the methods disclosed herein include alkaline phosphatases useful for converting phosphate-containing prodrugs into free drugs; aryl sulfatases useful for converting sulfate-containing prodrugs into free drugs; cytosine deaminases useful for converting non-toxic fluorocytosines into the anticancer drug 5-fluorouracil; proteases useful for converting peptide-containing prodrugs into free drugs, such as selatiana protease, thermolysin, subtilisin, carboxypeptidase and cathepsin (e.g., cathepsin B and L); and proteases containing D-amino acid substituents. The enzymes include, but are not limited to, D-alanyl carboxypeptidases useful for drug conversion; carbohydrate-cleaving enzymes such as O-galactosidases and neuraminidases useful for converting glycosylated prodrugs into free drugs; P-lactamases useful for converting drugs derivatized with P-lactams into free drugs; and penicillin amidases such as penicillin V amidase or penicillin G amidase, useful for converting drugs derivatized with their amine nitrogen groups having a phenoxyacetyl group or a phenylacetyl group, respectively, into free drugs. The enzymes can be covalently bonded to the polypeptides of this disclosure by techniques well known in the art, such as the use of the heterobifunctional crosslinking reagents described above.

[0161] According to one embodiment, in the complex of the present disclosure, the growth inhibitor is a maytansinoid, in one embodiment DM1 or DM4.

[0162] In the complex, the antibody is bound to at least one growth inhibitor by a binding group. In one embodiment, the binding group is a cleavable or incleavable linker such as SPDB, sulfo-SPDB, or SMCC.

[0163] In particular, anti-CEACAM5 antibody-drug conjugates can be selected from the following group: i) Equation (III): [ka] Anti-CEACAM5-SPDB-DM4 complex ii) Formula (IV): [ka] Anti-CEACAM5-sulfo-SPDB-DM4 complex and iii) Formula (V): [ka] The anti-CEACAM5-SMCC-DM1 complex.

[0164] In one embodiment, the immune complex is an immune complex of formula (III), (IV), or (V) as defined above, and the antibody is an antibody as described herein.

[0165] In certain embodiments, the immune complex comprises a heavy chain (VH) consisting of SEQ ID NO: 8 and a light chain (VL) consisting of SEQ ID NO: 9, and is mediated via N-succinimidyl pyridyldithiobutyrate (SPDB). 2 '-deacetyl-N 2 Contains an hCEACAM5 antibody covalently bound to (4-methyl-4-mercapto-1-oxopentyl)-meytansine (DM4).

[0166] In certain embodiments, the immune complex is tusamitamablutansin (CAS[2254086-60-5]).

[0167] Generally, immune complexes can be obtained by a process comprising: (i) contacting an optionally buffered aqueous solution of a cell binding agent (e.g., an antibody according to the present disclosure) with a solution of a linker and a cytotoxic compound; (ii) then optionally separating the complex formed in (i) from unreacted cell binding agent. The aqueous solution of the cell binding agent can be buffered with a buffer such as 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 dimethyl sulfoxide (DMSO) or dimethylacetamide (DMA).

[0168] The reaction temperature is usually 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 size exclusion chromatography (SEC) using a refractive index measurement and / or a UV detector. If the yield of the complex is too low, the reaction time can be extended.

[0169] For carrying out the separation in step (ii), several different chromatography methods can be used by those skilled in the art. The complex can be purified by, for example, SEC, adsorption chromatography (such as ion exchange chromatography, IEC, etc.), hydrophobic interaction chromatography (HIC), affinity chromatography, hydroxyapatite chromatography, or high performance liquid chromatography (HPLC). Purification by dialysis or diafiltration can also be used.

[0170] ​

[0171] As used herein, the term “aggregate” means an association that may be formed between two or more cell-binding agents, whether modified or unmodified by binding. Aggregates can form under the influence of numerous parameters, including high concentrations of cell-binding agents in solution, pH of the solution, high shear force, the number of bound dimers and their hydrophobicity, and temperature (see Wang & Gosh, 2008, J. Membrane Sci., 318:311-316 and the references cited therein). It should be noted that the relative effects of some of these parameters have not been clearly established. For proteins and antibodies, those skilled in the art will refer 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).

[0172] After step (i) or (ii), the complex-containing solution may be subjected to a further step (iii) of chromatography, ultrafiltration and / or diafiltration.

[0173] The complex is recovered in an aqueous solution at the end of these steps.

[0174] According to one embodiment, the complex according to the present disclosure is characterized by a “drug-to-antibody ratio” (or “DAR”) in the range of 1 to 10, for example, 2 to 5, and particularly 3 to 4. This is generally the case for a complex containing a meitansinoid molecule.

[0175] This DAR number can vary depending on the properties of the antibody and drug used (i.e., the growth inhibitor), along with the experimental conditions used for binding (such as the growth inhibitor / antibody ratio, reaction time, solvent properties, and co-solvent properties, if present). Therefore, contact between the antibody and growth inhibitor results in several different complexes depending on the drug-to-antibody ratio; optionally, naked antibodies; optionally, mixtures containing aggregates. Thus, the determined DAR is an average value.

[0176] The method that can be used to determine DAR is λ D This involves measuring the ratio of absorbances of a substantially purified complex solution at 280 nm using a spectrophotometer. 280 nm is a wavelength commonly used for measuring protein concentrations, such as antibody concentrations. D The antibodies are selected to allow for the identification of drugs, that is, as is readily known to those skilled in the art, λ D The drug has high absorbance, λ D λ is a wavelength sufficiently far from 280 nm to avoid substantial overlap of the absorbance peaks of the drug and antibody. D For maytansinoid molecules, this can be selected as 252 nm. The method for DAR calculation can be obtained from Antony S. Dimitrov (ed), LLC, 2009, Therapeutic Antibodies and Protocols, vol 525, 445, Springer Science.

[0177] λ D (A λD ) and 280nm(A 280 The absorbance of the complex at ) is measured using either a monomer peak from size exclusion chromatography (SEC) analysis (which allows for the calculation of the "DAR(SEC)" parameter) or a classical spectrophotometer (which allows for the calculation of the "DAR(UV)" parameter). The absorbance can be expressed as follows: A λ D = (cD × εD) λ D) + (cA × εA λ D) A 280 =(cD×εD 280 ) + (cA × εA 280 ) (In the formula, cD and cA are the concentrations of the drug (i.e., chemotherapeutic agent) and antibody in solution, respectively. εD λ D and εD 280 These are the molar extinction coefficients of the drug at λD and 280 nm, respectively. εA λ D and εA 280 (These are the molar extinction coefficients of the antibody at λD and 280 nm, respectively.)

[0178] The resolution of these two equations, which have two unknowns, yields the following equation: cD=[(εA 280 ×A λ D)-(εA λ D×A 280 )] / [(εD λ D×εA 280 )-(εA λ D×εD 280 )] cA = [A 280 -(cD×εD 280 )] / εA 280

[0179] Next, the average DAR is calculated from the ratio of the antibody concentration to the drug concentration: DAR = cD / cA.

[0180] In certain embodiments, the immune complex comprises a heavy chain consisting of SEQ ID NO: 8 and a light chain consisting of SEQ ID NO: 9, and is mediated via N-succinimidyl pyridyldithiobutyrate (SPDB). 2 '-deacetyl-N 2 Contains an hCEACAM5 antibody covalently bound to (4-methyl-4-mercapto-1-oxopentyl)-meytansine (DM4).

[0181] In certain embodiments, the immune complex is tusamitamaburabutansin.

[0182] Neuroendocrine cancer The present disclosure relates to an antibody, or an antigen-binding fragment thereof, or an immune complex comprising the antibody or the antigen-binding fragment, for use in treating a cancer selected from neuroendocrine cancers that express CEACAM5 in a subject in need thereof.

[0183] This further relates to a method for treating a cancer selected from neuroendocrine cancers that express hCEACAM5 in a subject in need thereof, the method comprising administering an antibody, or an immune complex comprising the antibody.

[0184] Neuroendocrine cancer is a neoplasm that arises from cells of the endocrine and nervous systems. Two major histological subtypes of neuroendocrine cancer have been identified: small cell type and large cell type. Neuroendocrine cancer can occur in almost any body site. Small cell neuroendocrine cancer of the lung is the most frequent subset, with others being rare or very rare. Neuroendocrine cancer is usually diagnosed at an advanced metastatic stage.

[0185] As demonstrated by the recent approvals of brentuximab vedotin for the treatment of Hodgkin lymphoma and trastuzumab emtansine (T-DM1) for the treatment of recurrent metastatic HER2+ breast cancer, the use of ADCs to selectively target potent cytotoxic agents to tumor cells has now been shown to be an effective strategy for cancer treatment. 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 sufficiently expressed on tumor cells to achieve selective and efficient uptake of the drug.

[0186] The neuroendocrine cancers of the present disclosure are selected from neuroendocrine tumors (NETs), neuroendocrine carcinomas (NECs), mixed neuroendocrine-non-neuroendocrine neoplasms (MiNENs), pheochromocytomas, and medullary thyroid cancer (MTC).

[0187] In certain embodiments, cancer is - Neuroendocrine cancers of the gastrointestinal tract and pancreatic and biliary ducts, such as neuroendocrine cancers of the digestive tract, especially the esophagus, stomach, pancreas, liver, small intestine, and large intestine or anal region, or neuroendocrine cancers of the urinary tract, especially neuroendocrine cancers of the bladder. - Neuroendocrine cancers of the upper respiratory tract, gastrointestinal tract, and salivary glands, including neuroendocrine cancers of the lung, particularly small cell lung cancer, and neuroendocrine cancers of the thymus. -Medullary thyroid carcinoma (MTC) and other neuroendocrine cancers of the thyroid gland, - Neuroendocrine cancers of the adrenal gland, such as pheochromocytoma, - Neuroendocrine cancers of the skin, such as Merkel cell carcinoma (MCC) - Neuroendocrine cancer of the female reproductive organs such as the endometrium, cervix, or ovaries. - Neuroendocrine cancers of the prostate or testes, especially the testes and other male reproductive organs, and - Neuroendocrine cancer of the head and neck Selected from.

[0188] In certain embodiments, neuroendocrine cancers are selected from neuroendocrine cancers of the esophagus, stomach, small and large intestine, anal region, pancreas, bladder, lung, female reproductive organs, male reproductive organs, and head and neck.

[0189] In certain embodiments, neuroendocrine cancer is neuroendocrine cancer of the lung, particularly neuroendocrine small cell lung cancer, or neuroendocrine cancer of the male reproductive organs, particularly neuroendocrine prostate cancer.

[0190] In certain embodiments, neuroendocrine cancer is neuroendocrine cancer of the lung, particularly neuroendocrine small cell lung cancer.

[0191] In certain embodiments, neuroendocrine cancer is not neuroendocrine small cell lung cancer. In particular, neuroendocrine cancer is selected from the group consisting of neuroendocrine cancers of the esophagus, stomach, small and large intestine, anal region, pancreas, bladder, female reproductive organs, male reproductive organs, thyroid gland, and head and neck.

[0192] In certain embodiments, neuroendocrine cancer is neuroendocrine prostate cancer (NEPC).

[0193] In certain embodiments, neuroendocrine carcinoma is not prostate cancer. In particular, neuroendocrine carcinoma is not NEPC. In particular, neuroendocrine carcinoma is selected from the group consisting of neuroendocrine carcinomas of the esophagus, stomach, small and large intestine, anal region, pancreas, bladder, lung, female reproductive organs, thyroid, and head and neck.

[0194] Neuroendocrine subjects treated in accordance with this disclosure may be classified as moderately or highly CEACAM5 expressors.

[0195] In one embodiment, the subjects are individuals who express moderate or high levels of carcinoembryonic antigen-associated cell adhesion molecules.

[0196] In a particular embodiment, the subjects are moderate CEACAM5 expressors. Moderate CEACAM5 expressors have CEACAM5 expression of +2 or 3+ intensity in at least 1% of the tumor cell population, or CEACAM5 expression of 1+ intensity in at least 50% of the tumor cell population, as measured by hCEACAM5 immunohistochemistry.

[0197] In a particular embodiment, the subjects are high-CEACAM5 expressors. High-CEACAM5 expressors have CEACAM5 expression of 2+ or 3+ intensity in more than 50% of the tumor cell population, as measured by hCEACAM5 immunohistochemistry.

[0198] In certain embodiments, the disclosure further relates to antibodies or antigen-binding fragments thereof, or immunocomplexes comprising such antibodies or antigen-binding fragments, for use as pharmaceuticals.

[0199] In certain embodiments, the Disclosure relates to the use of an antibody, or an antigen-binding fragment thereof, or an immune complex comprising such antibody or antigen-binding fragment, for the manufacture of a pharmaceutical product for treating selected cancers from neuroendocrine cancers that express CEACAM5 in a subject where such treatment is necessary.

[0200] The neuroendocrine cancer may be selected from any of those described above in this disclosure.

[0201] In certain embodiments, the Disclosure relates to an immune complex comprising an antibody or an antigen-binding fragment thereof for use in treating cancers selected from neuroendocrine cancers expressing hCEACAM5 in subjects where such treatment is necessary, wherein the antibody specifically binds to hCEACAM5, and the antibody or its antigen-binding fragment comprises a VH domain and a VL domain, the VH domain comprising three complementarity-determining regions (CDRs) HCDR1, HCDR2, and HCDR3, and the VL domain comprising three CDRs LCDR1, LCDR2, and LCDR3. HCDR1 contains the amino acid sequence of SEQ ID NO: 3 (GFVFSSYD), HCDR2 contains the amino acid sequence of SEQ ID NO: 4 (ISSGGGIT), and HCDR3 contains the amino acid sequence of SEQ ID NO: 5 (AAHYFGSSGPFAY). LCDR1 contains the amino acid sequence of SEQ ID NO: 6 (ENIFSY), LCDR2 contains the amino acid sequence of NTR, and LCDR3 contains the amino acid sequence of SEQ ID NO: 7 (QHHYGTPFT). The antibody is conjugated or linked to at least one growth inhibitor, the at least one growth inhibitor being selected from the group consisting of chemotherapeutic agents, enzymes, antibiotics and small molecule toxins or enzymatically active toxins, taxoids, vinca, taxanes, maytansinoids or maytansinoid analogs, tomaimycin or pyrrolobenzodiazepine derivatives, cryptophycin derivatives, leptomycin derivatives, auristatin or drastatin analogs, prodrugs, topoisomerase II inhibitors, DNA alkylating agents, antitubulin agents and CC-1065 or CC-1065 analogs, particularly excluding topoisomerase I inhibitors.

[0202] In certain embodiments, the Disclosure relates to an immune complex comprising an antibody or an antigen-binding fragment thereof for use in treating cancers selected from neuroendocrine cancers expressing hCEACAM5 in subjects where such treatment is necessary, wherein the antibody specifically binds to hCEACAM5, and the antibody or its antigen-binding fragment comprises a VH domain and a VL domain, the VH domain comprising three complementarity-determining regions (CDRs) HCDR1, HCDR2, and HCDR3, and the VL domain comprising three CDRs LCDR1, LCDR2, and LCDR3. HCDR1 contains the amino acid sequence of SEQ ID NO: 3 (GFVFSSYD), HCDR2 contains the amino acid sequence of SEQ ID NO: 4 (ISSGGGIT), and HCDR3 contains the amino acid sequence of SEQ ID NO: 5 (AAHYFGSSGPFAY). LCDR1 contains the amino acid sequence of SEQ ID NO: 6 (ENIFSY), LCDR2 contains the amino acid sequence of NTR, and LCDR3 contains the amino acid sequence of SEQ ID NO: 7 (QHHYGTPFT). The antibody is conjugated or linked to at least one growth inhibitor, the at least one growth inhibitor being selected from the group consisting of chemotherapeutic agents, enzymes, antibiotics and small molecule toxins or enzymatically active toxins, taxoids, vinca, taxanes, maytansinoids or maytansinoid analogs, tomaimycin or pyrrolobenzodiazepine derivatives, cryptophycin derivatives, leptomycin derivatives, auristatin or drastatin analogs, prodrugs, topoisomerase inhibitors, DNA alkylating agents, antitubulin agents and CC-1065 or CC-1065 analogs. Neuroendocrine cancer is selected from the group consisting of neuroendocrine cancers of the esophagus, stomach, small and large intestine, anal region, pancreas, bladder, female reproductive organs, male reproductive organs, thyroid gland, and head and neck, and in particular, this neuroendocrine cancer is not neuroendocrine small cell lung cancer (SCLC).

[0203] Pharmaceutical composition The antibodies or immune complexes of this disclosure can be combined with pharmaceutically acceptable excipients and optionally with a sustained-release matrix (such as a biodegradable polymer) to form therapeutic compositions.

[0204] Therefore, another object of this disclosure relates to a pharmaceutical composition comprising an antibody or immune complex of this disclosure and a pharmaceutically acceptable carrier or excipient.

[0205] This disclosure also relates to polypeptides or immune complexes relating to this disclosure for use as pharmaceuticals.

[0206] "Pharmacologically" or "pharmaceutically acceptable" means molecular entities and compositions that, as appropriate, do not cause adverse allergic reactions or other undesirable reactions when administered to mammals, particularly humans. A pharmacopoeia is a pharmacopoeia of any kind of non-toxic solid, semi-solid, or liquid filler, diluent, encapsulating material, or formulation aid.

[0207] As used herein, “pharmaceutically acceptable carrier” includes any physiologically compatible solvent, dispersion medium, coating, antimicrobial agent, and antifungal agent. Examples of suitable carriers, diluents, and / or excipients include water, amino acids, physiological saline, phosphate-buffered saline, phosphate buffer, acetate, citrate, succinate; amino acids and derivatives such as histidine, arginine, glycine, proline, and glycylglycine; inorganic salts such as NaCl and calcium chloride; sugars or polyalcohols such as dextrose, glycerol, ethanol, sucrose, trehalose, and mannitol; surfactants such as polysorbate 80, polysorbate 20, and poloxamer 188, and one or more combinations thereof. Often, it is preferable to include isotonic agents, such as sugars, polyalcohols, or sodium chloride in the composition, and the formulation may also contain antioxidants, such as tryptamine, and stabilizers, such as Tween 20.

[0208] The form, route of administration, dosage, and regimen of a pharmaceutical composition naturally depend on the condition being treated, the severity of the disease, the patient's age, weight, and sex, etc.

[0209] The pharmaceutical compositions of this disclosure can be formulated for topical, oral, parenteral, intranasal, intravenous, intramuscular, subcutaneous, or intraocular administration.

[0210] The therapeutic compositions disclosed herein are administered with suitable carriers, excipients, and other agents incorporated into the formulation to provide improved transport, delivery, tolerability, etc. Numerous suitable formulations can be found in the prescription book known to all medicinal chemists: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, which is incorporated herein by reference in its entirety. Examples of these formulations include powders, pastes, ointments, jellies, waxes, oils, lipids, lipid (cationic or anionic)-containing vesicles (e.g., LIPOFECTIN®), DNA complexes, anhydrous absorbent pastes, oil-in-water and water-in-oil emulsions, emulsion carbowaxes (polyethylene glycol of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowaxes. See also Powell et al. “Compendium of excipients for parenteral formulations” PDA (1998) J Pharm Sci Technol 52:238-311, which is incorporated herein by reference in its entirety.

[0211] Various delivery systems are known and can be used to administer the pharmaceutical compositions of this disclosure, for example, liposomes, microparticles, microcapsules, and encapsulation in receptor-mediated endocytosis (see, for example, Wu et al. (1987) J. Biol. Chem. 262:4429-4432, which is incorporated herein in its entirety by reference). Methods of delivery include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions may be administered by any convenient route, for example, by infusion or bolus injection, by absorption via epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa), and may be administered together with other bioactive agents. Administration may be systemic or topical. CEACAM5 antibodies, or antibody-containing immune complexes, may be administered subcutaneously.

[0212] Pharmaceutical compositions can also be delivered in vesicles such as liposomes (see Langer (1990) Science 249:1527-1533, the whole of which is incorporated herein by reference). In certain circumstances, pharmaceutical compositions can be delivered by controlled release systems, for example, using pumps or polymer materials. In another embodiment, the controlled release system may be positioned near the target of the composition, thus requiring only a portion of the systemic dose.

[0213] Injectable formulations include dosage forms for intravenous, subcutaneous, intradermal, and intramuscular injection, local injection, and intravenous infusion. These injectable formulations can be prepared by known methods. For example, an injectable formulation may be prepared by dissolving, suspending, or emulsifying the antibody or a salt thereof in a sterile aqueous or oily medium conventionally used for injection. Examples of aqueous media for injection include physiological saline, glucose, and other adjuvants, which may be used in combination with appropriate solubilizers such as alcohol (e.g., ethanol), polyalcohol (e.g., propylene glycol, polyethylene glycol), and nonionic surfactants (e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)). Examples of oily media include sesame oil and soybean oil, which may be used in combination with solubilizers such as benzyl benzoate and benzyl alcohol. The injection thus prepared can be filled into appropriate ampoules.

[0214] Advantageously, the above-mentioned pharmaceutical compositions for oral or parenteral use are prepared into dosage forms of unit doses suitable for the dosage of the active ingredient. Such dosage forms of unit doses include, for example, tablets, pills, capsules, injections (ampoules), and suppositories.

[0215] In one embodiment, the pharmaceutical composition contains a pharmaceutically acceptable vehicle for an injectable formulation. These may be dry compositions, particularly lyophilized compositions, that enable the formation of an injectable solution by adding isotonic sterile saline (monosodium or disodium phosphate, sodium chloride, potassium, calcium or magnesium, etc., or mixtures of such salts) or optionally sterile water or saline.

[0216] The pharmaceutical composition may be administered through a drug combination device.

[0217] The dosage used for administration can be adapted as a function of various parameters, such as the mode of administration used, the relevant disease state, or the desired duration of treatment.

[0218] For the preparation of a pharmaceutical composition, an effective amount of the antibody or immune complex of this disclosure can be dissolved or dispersed in a pharmaceutically acceptable carrier or aqueous medium.

[0219] Suitable pharmaceutical forms for injectable use include sterile aqueous solutions or dispersions, formulations containing sesame oil, peanut oil, or aqueous propylene glycol, and sterile powders for the immediate preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and injectable using a suitable device or system for delivery without decomposition. It must be stable under manufacturing and storage conditions and protected from contamination by microorganisms such as bacteria and fungi.

[0220] Solutions of the active compound, as a free base or a pharmacokinetically acceptable salt, can be prepared in water, which is suitably mixed with a surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycol, and mixtures thereof, as well as in oil. Under normal storage and use conditions, these preparations contain preservatives to prevent microbial growth.

[0221] The antibodies or immune complexes of this disclosure can be formulated into compositions in neutral or salt form. Pharmaceutically acceptable salts include acid addition salts (formed with free amino groups of proteins) formed with inorganic acids, such as hydrochloric acid or phosphoric acid, or organic acids such as acetic acid, oxalic acid, tartaric acid, or mandelic acid. Salts formed with free carboxyl groups can also be derived from inorganic bases, such as sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide, or ferric hydroxide, and organic bases, such as isopropylamine, trimethylamine, glycine, histidine, or procaine.

[0222] The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oils. Appropriate fluidity can be maintained, for example, by the use of coating materials such as lecithin, maintaining the required particle size in the case of dispersions, and the use of surfactants. Prevention of microbial action can be achieved by various antimicrobial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, and thimerosal. In many cases, it is preferable to include isotonic agents, such as sugars or sodium chloride. Sustained absorption of the injectable composition can be achieved by the use of absorption-delaying agents (e.g., aluminum monostearate and gelatin) in the composition.

[0223] Sterile injectable solutions are prepared by incorporating the required amount of active compound, along with any other components listed above as needed, into a suitable solvent, followed by sterilization by filtration. Generally, dispersions are prepared by incorporating various sterilized active ingredients into a sterile vehicle containing a basic dispersion medium and other necessary components from those listed above. For sterile powders for the preparation of sterile injectable solutions, preferred preparation methods are vacuum drying and freeze-drying techniques, which yield a powder of the active ingredient + any additional desired components from its previously sterilized filtered solution.

[0224] The preparation of more concentrated or higher-concentration solutions for direct injection is also being considered, and the use of DMSO as a solvent is expected to result in extremely rapid penetration, allowing for the delivery of high concentrations of the active agent to small tumor areas.

[0225] Once formulated, the solution is administered in a therapeutically effective amount in a form compatible with the administered preparation. The preparation can be easily administered in various dosage forms, such as the types of injection solutions mentioned above, but drug-releasing capsules and the like can also be used.

[0226] For parenteral administration in aqueous solutions, for example, the solution should be buffered as needed, and the liquid diluent should first be isotonic with sufficient saline or glucose. These aqueous solutions are particularly suitable for intravenous, intramuscular, subcutaneous, and intraperitoneal administration. In this regard, the sterile aqueous media that can be used will be known to those skilled in the art in light of this disclosure. For example, one dose can be dissolved in 1 ml of isotonic NaCl solution and added to 1000 ml of subcutaneous injection solution, or injected into the proposed injection site (e.g., “Remington's Pharmaceutical Sciences” 15 th (See Edition, pages 1035-1038 and 1570-1580). Some variation in dosage will inevitably occur depending on the condition of the subject being treated. The person responsible for administration will, in any case, determine the appropriate dose for each individual subject.

[0227] The antibodies or immune complexes of this disclosure may be formulated in therapeutic mixtures to contain approximately 0.01 to 100 milligrams / dose, etc.

[0228] In addition to antibodies or immune complexes formulated for parenteral administration, such as intravenous or intramuscular injection, other pharmaceutically acceptable forms include, for example, tablets or other solids for oral administration, time-release capsules, and any other forms currently in use.

[0229] In certain embodiments, the use of liposomes and / or nanoparticles is intended for the introduction of polypeptides into host cells. The formation and use of liposomes and / or nanoparticles are known to those skilled in the art.

[0230] Nanocapsules can generally capture compounds in a stable and reproducible manner. To avoid side effects due to intracellular polymer overload, such ultrafine particles (approximately 0.1 μm in size) are generally designed using polymers that can be degraded in vivo. Biodegradable polyalkyl-cyanoacrylate nanoparticles, or biodegradable polylactide or polylactide coglycolide nanoparticles that meet these requirements, are intended for use in this disclosure, and such particles can be readily fabricated.

[0231] Liposomes are formed from phospholipids that disperse in an aqueous medium and spontaneously form multilayer concentric bilayer vesicles (also called multilayer vesicles (MLVs)). MLVs generally have a diameter of 25 nm to 4 μm. Sonic treatment of MLVs results in the formation of small unilamellar vesicles (SUVs) with a diameter ranging from 200 to 500 Å, containing an aqueous solution in the core. The physical characteristics of liposomes depend on pH, ionic strength, and the presence of divalent cations.

[0232] Method of administration The methods described herein include administering a therapeutically effective dose of an anti-CEACAM5 antibody or an antibody-containing immune complex to a target.

[0233] As used herein, “effective dose” or “therapeutic effective dose” refers to the dose of a therapeutic agent that results in treatment of neuroendocrine cancer. As used herein, “treatment” means resulting in a detectable improvement in one or more symptoms associated with neuroendocrine cancer or in a biological effect that correlates with the underlying pathological mechanism causing the condition or symptoms (e.g., a reduction in the level of a particular biomarker). For example, a dose of an anti-CEACAM5 antibody or an antibody-containing immune complex that results in improvement in any of the symptoms or conditions associated with neuroendocrine cancer is considered a “therapeutic effective dose.”

[0234] In another example, the treatment was deemed ineffective when the dose of anti-CEACAM5 antibody or an antibody-containing immune complex did not result in a detectable improvement in one or more parameters or symptoms associated with neuroendocrine cancer, or when it did not produce a biological effect correlated with one or more pathological mechanisms underlying such cancerous conditions or symptoms.

[0235] According to some of these embodiments, the anti-CEACAM5 antibody or the antibody-containing immune complex is administered intravenously.

[0236] According to the method of this disclosure, the therapeutically effective dose of anti-CEACAM5 antibody or antibody-containing immune complex administered to a subject varies depending on the subject's age and size (e.g., body weight or body surface area), the route of administration, and other factors well known to those skilled in the art.

[0237] The treatment or course of treatment may include at least one treatment cycle.

[0238] In some embodiments, the treatment may include a first cycle of treatment, i.e., cycle 1, and at least one additional cycle of treatment, i.e., cycles 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or more.

[0239] The first cycle and the additional cycles may be the same or different.

[0240] For example, the first cycle may include the administration of a loading dose, and one or more additional cycles may include the administration of a maintenance dose.

[0241] Alternatively, the first cycle and any additional cycles may include administration of the same dose.

[0242] In some embodiments, the antibody-drug conjugate containing an anti-CEACAM5 antibody may be administered in a loading dose for the first cycle and a maintenance dose for one or more additional cycles.

[0243] In some embodiments, the antibody-drug conjugate containing an anti-CEACAM5 antibody may be administered in the same dose for the first cycle and one or more additional cycles.

[0244] Treatment cycles can last approximately 1 to 6 weeks, 1 to 4 weeks, or 1 to 3 weeks.

[0245] In some embodiments, the treatment cycle can last for at least about two weeks.

[0246] In some embodiments, the treatment cycle can last for at least about three weeks.

[0247] In some embodiments, a treatment cycle may include a treatment period of at least one day of the cycle, for example, day 1, day 2, day 3, day 4, day 5, or day 6, and a rest period that continues until the completion of the cycle. The treatment period and the rest period may be the same or different between the first cycle and at least one additional cycle. In some embodiments, the treatment period and the rest period may be the same between the first cycle and at least one additional cycle.

[0248] In some embodiments, the treatment cycle may include a treatment period on the first day of the cycle and a rest period that continues until the completion of the cycle.

[0249] The treatment (or course of treatment) may include at least one first cycle of treatment (cycle 1) and at least one further (subsequent) cycle. The treatment may include 2-16, 3-15, 4-14, 5-13, 6-12, 7-11, 8-10, or about 9 cycles. The treatment may include 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or more cycles.

[0250] In certain embodiments, the dosage of an antibody or an immune complex comprising an antibody varies according to the body surface area of the subject. In certain embodiments, the dosage of the anti-CEACAM5 antibody or immune complex comprising an antibody administered to the subject is about 1 mg / m 2 ~ about 500 mg / m 2 . In some embodiments, the dosage of the antibody or immune complex comprising an antibody administered to the subject is about 5 mg to about 300 mg / m 2 . In various embodiments, the dosage of the antibody or immune complex comprising an antibody administered to the subject is about 5 to about 250 mg / m 2 .

[0251] In certain embodiments, the antibody or immune complex is administered at a dosage level of 5, 10, 20, 30, 40, 60, 80, 100, 120, 150, 180 or 210 mg / m 2 based on the body surface area of the subject. In various embodiments, the antibody or immune complex comprising an antibody is administered at a dosage of about 2.5 mg / m 2 ~ about 5 mg / m 2 . For example, the antibody or immune complex comprising an antibody is administered at a dosage of about 2.5 mg / m 2 ~ about 5 mg / m 2 over a period of time (e.g., 30 minutes and 1 hour). The dosage includes 2.5 mg / m 2 of the antibody, 5 mg / m 2 of the antibody, or an immune complex comprising an antibody, and all dosages from 2.5 mg / m 2 to 5 mg / m 2 , e.g., 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8 and 4.9 mg / m 2 .

[0252] For example, the present disclosure provides about 1 mg / m 2 , about 5 mg / m 2 , 10 mg / m 2 , about 15 mg / m 2 , about 20 mg / m 2 , about 25 mg / m 2 , about 30 mg / m2 Approximately 35 mg / m 2 Approximately 40 mg / m 2 Approximately 45mg / m 2 Approximately 50 mg / m 2 Approximately 55mg / m 2 Approximately 60 mg / m 2 Approximately 65mg / m 2 Approximately 70 mg / m 2 Approximately 75mg / m 2 Approximately 80 mg / m 2 Approximately 85 mg / m 2 Approximately 90 mg / m 2 Approximately 95mg / m 2 Approximately 100 mg / m 2 Approximately 105 mg / m 2 Approximately 110 mg / m 2 Approximately 115 mg / m 2 Approximately 120 mg / m 2 Approximately 125 mg / m 2 Approximately 130 mg / m 2 Approximately 135 mg / m 2 Approximately 140 mg / m 2 Approximately 145 mg / m 2 Approximately 150 mg / m 2 Approximately 155 mg / m 2 Approximately 160 mg / m 2 Approximately 165 mg / m 2 Approximately 170 mg / m 2 Approximately 175 mg / m 2 Approximately 180 mg / m 2 Approximately 185 mg / m 2 Approximately 190 mg / m 2 Approximately 195 mg / m 2 Approximately 200 mg / m 2 Approximately 205 mg / m 2 Approximately 210 mg / m 2 Approximately 215 mg / m 2 Approximately 220 mg / m 2 Approximately 225 mg / m 2 Approximately 230 mg / m 2 Approximately 235 mg / m 2 Approximately 240 mg / m 2 Approximately 245 mg / m 2 Approximately 250 mg / m 2 Approximately 255 mg / m 2 Approximately 260 mg / m2 , about 265mg / m 2 , about 270mg / m 2 , about 275mg / m 2 , about 280mg / m 2 , about 285mg / m 2 , about 290mg / m 2 , about 295mg / m 2 , about 300mg / m 2 , about 325mg / m 2 , about 350mg / m 2 , about 375mg / m 2 , about 400mg / m 2 , about 425mg / m 2 , about 450mg / m 2 , about 475mg / m 2 Or approximately 500 mg / m² 2 The anti-CEACAM5 antibody, or an immune complex containing the antibody, is administered to the patient once per week or every two weeks, including, but not limited to, a method of administration.

[0253] In various embodiments, the antibody or immune complex is administered at a dose of 5, 10, 20, 30, 40, 60, 80, 100, 120, 150, 180, or 210 mg / m² based on the target body surface area. 2 It is administered at this dose level.

[0254] In certain embodiments, the antibody or immune complex is administered every 14 days (2 weeks) or every 3 weeks.

[0255] In certain embodiments, the antibody or immune complex is administered at a dose level of 100 mg / m2 every two weeks, based on the target body surface area.

[0256] As used herein, “administered at approximately 1 to approximately 500 mg / kg” means that the referenced substance is administered at any value within the stated range, including the range endpoint. For example, “the dose of anti-CEACAM5 antibody or antibody-containing immune complex administered to a patient is 1 mg / m².” 2 ~500mg / m 2 "is" is 1 mg / m² 2Anti-CEACAM5 antibody or antibody-containing immune complex, 500 mg / m² 2 The administration of an anti-CEACAM5 antibody or an antibody-containing immune complex and all doses therein is included. In one embodiment, the CEACAM5 antibody or antibody-containing immune complex is administered once over a certain period, for example, every 14 days (i.e., every 2 weeks) or every 3 weeks, at doses of approximately 5, 10, 20, 30, 40, 60, 80, 100, 120, 150, 180, or 210 mg / m². 2 It is administered in the following dosage.

[0257] In various embodiments, the dose is administered at a constant rate, or at a variable rate. In various embodiments, the dose is administered at a constant rate of approximately 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 2.5, or 5 mg / min. In various embodiments, the antibody or antibody-containing immune complex is administered at a rate over the first 30 minutes or 1 hour. In various embodiments, after approximately 30 minutes or 1 hour, the rate of antibody administration is changed. For example, the rate is decreased. In various embodiments, the rate is increased. In various embodiments, the antibody or antibody-containing immune complex is administered at a rate of 2.5 mg / min over the first 30 minutes or 1 hour. In various embodiments, after approximately 30 minutes or 1 hour, the rate of antibody or antibody-containing immune complex administration is increased to 5 mg / min.

[0258] The method of the present disclosure comprises administering multiple doses of an anti-CEACAM5 antibody or an antibody-containing immune complex to a patient over a specified period of time. For example, the anti-CEACAM5 antibody or antibody-containing immune complex may be administered approximately 1 to 5 times per day, approximately 1 to 5 times per week, approximately 1 to 5 times every two weeks, approximately 1 to 5 times every month, or approximately 1 to 5 times every year. In certain embodiments, the method of the present disclosure comprises administering a first dose of the anti-CEACAM5 antibody or antibody-containing immune complex to a patient at a first time point, followed by administering at least a second dose of the anti-CEACAM5 antibody or antibody-containing immune complex to the patient at a second time point. In certain embodiments, the first and second doses may contain the same amount of anti-CEACAM5 antibody or antibody-containing immune complex. The time between the first and second doses may range from approximately several hours to several weeks. For example, the second time point (i.e., the time when the second dose is administered) may be about 1 hour to about 7 weeks after the first time point (i.e., the time when the first dose is administered). According to certain exemplary embodiments of the Disclosure, the second time point may be about 1 hour, about 4 hours, about 6 hours, about 8 hours, about 10 hours, about 12 hours, about 24 hours, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 2 weeks, about 3 weeks, about 4 weeks, about 6 weeks, about 8 weeks, about 10 weeks, about 12 weeks, or about 14 weeks or more after the first time point. In certain embodiments, the second time point is about 1 week or about 2 weeks. The third dose and subsequent doses may be administered similarly throughout the patient's treatment course. The Disclosure provides a method of using a therapeutic composition comprising an anti-CEACAM5 antibody or its antigen-binding fragment, or an immune complex containing the antibody, and optionally one or more further therapeutic agents.

[0259] According to one embodiment, the antibody or immune complex is administered as a loading dose of 80, 100, 120, 135, 150, or 170 mg / m². 2 It is administered in the following dosage.

[0260] In this disclosure, the term “loading dose” is intended to refer to the dose of the drug used at the initiation of a procedure to front-load an appropriate plasma concentration of the drug that is subsequently maintained by a maintenance dose. The loading dose is typically higher than the maintenance dose. “Maintenance dose” is intended to refer to the dose of the drug administered on a regular schedule to maintain a plateau in plasma drug concentration once a high plasma concentration of the drug has been established by the use of a loading dose. The maintenance dose is typically lower than the loading dose.

[0261] According to one embodiment, the antibody or immune complex is administered as a loading dose of 120, 135, 150, or 170 mg / m². 2 It is administered in the following dosage.

[0262] In certain embodiments, the antibody or immune complex is administered as a loading dose of approximately 100 mg / m². 2 ~about 200mg / m 2 In particular, the loading dose is approximately 135 mg / m². 2 , about 150mg / m 2 , or 170 mg / m² 2 It is administered in the following dosage.

[0263] According to one embodiment, the antibody or immune complex is administered as a maintenance dose of 100 mg / m². 2 It is administered in the following dosage.

[0264] According to one embodiment, the antibody or immune complex is administered as a loading dose of 120, 135, 150, or 170 mg / m² in cycle 1. 2 The dose is 100 mg / m², followed by a maintenance dose of 100 mg / m² in one or more further cycles. 2 It is administered in the following dosage.

[0265] According to one embodiment, the antibody or immune complex is 150 mg / m² in cycle 1. 2 It is administered at this dose level, and the cycle is approximately two weeks.

[0266] According to one embodiment, the antibody or immune complex is administered as a loading dose of 150 mg / m² in cycle 1.2 The dose is 100 mg / m², followed by a maintenance dose of 100 mg / m² in one or more further cycles. 2 It is administered in this dose. The cycle may be approximately 2 weeks.

[0267] According to one embodiment, the antibody or immune complex is 170 mg / m² in cycle 1. 2 It is administered in this dose, and the cycle is approximately 2 or 3 weeks.

[0268] According to one embodiment, the antibody or immune complex is administered as a loading dose of 170 mg / m² in cycle 1. 2 For the following doses, the maintenance dose will be 100 mg / m² for one or more further cycles. 2 It is administered in the specified dose. The cycle may be approximately 2 or 3 weeks.

[0269] According to one embodiment, the antibody or immune complex is administered at a dose of 100 mg / m² in all cycles, i.e., cycle 1 and one or more further cycles. 2 It is administered in the specified dose. The cycle may be approximately 2 or 3 weeks.

[0270] According to the methods disclosed in this invention, an anti-CEACAM5 antibody or an antibody-containing immune complex (or a pharmaceutical formulation containing an antibody or an antibody-containing immune complex) can be administered to a patient using any acceptable device or mechanism. For example, administration can be achieved using a syringe and needle, or using a reusable pen and / or auto-injector delivery device. The methods of this disclosure include the use of a number of reusable pens and / or auto-injector delivery devices for administering an anti-CEACAM5 antibody or an antibody-containing immune complex (or a pharmaceutical formulation containing an antibody or an antibody-containing immune complex). Examples of such devices include, but are not limited to, AUTOPEN (Owen Mumford, Inc. (Woodstock, UK)), DISETRONIC pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75 / 25 pen, HUMALOG pen, HUMALIN 70 / 30 pen (Eli Lilly and Co., Indianapolis, IN), NOVOPENI, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR (Novo Nordisk, Copenhagen, Denmark), BD pen (Becton Dickinson, Franklin Lakes, NJ), OPTIPEN, OPTIPEN PRO, OPTIPEN STARLET and OPTICLIK (Sanofi-Aventis, Frankfurt, Germany). Examples of disposable pens and / or auto-injector delivery devices for use in subcutaneous delivery of the pharmaceutical compositions of this disclosure include, but are not limited to, SOLOSTAR pens (Sanofi-Aventis), FLEXPEN (Novo Nordisk), and KWIKPEN (Eli Lilly), SURECLICK Autoinjector (Amgen, Thousand Oaks, CA), PENLET (Haselmeier, Stuttgart, Germany), EPIPEN (Dey, LP), and HUMIRAPen (AbbVie Inc., North Chicago, IL).

[0271] In one embodiment, the antibody or an antibody-containing immune complex is administered in a pre-filled syringe. In another embodiment, the antibody or an antibody-containing immune complex is administered in a pre-filled syringe containing a safety system, for example, to prevent injury from accidental needle sticks. In various embodiments, the antibody is administered in a pre-filled syringe containing the ERIS safety system (West Pharmaceutical Services Inc.). See also U.S. Patents No. 5,215,534 and No. 9,248,242, which are incorporated herein by reference in their entirety.

[0272] In another embodiment, the antibody or antibody-containing immune complex is administered by auto-injector. In various embodiments, the antibody or antibody-containing immune complex is administered by auto-injector featuring PUSHCLICK technology (SHL Group). In various embodiments, the auto-injector is a device comprising a syringe that enables the administration of a dose of the composition and / or antibody to a subject. See also U.S. Patent Nos. 9,427,531 and 9,566,395, which are incorporated herein by reference in their entirety.

[0273] The use of microinjectors for delivering anti-CEACAM5 antibodies or antibody-containing immune complexes (or pharmaceutical formulations containing antibodies or antibody-containing immune complexes) to patients is also contemplated herein. As used herein, the term “microinjector” means a subcutaneous delivery device designed to slowly administer a large volume of therapeutic formulation (e.g., up to approximately 2.5 mL or more) over an extended period (e.g., approximately 10, 15, 20, 25, 30 minutes or more). See, for example, U.S. Patent No. 6,629,949, U.S. Patent No. 6,659,982 and Meehan et al., J. Controlled Release 46:107-116 (1996), which are incorporated herein by reference in their entirety. Microinjectors are particularly useful for delivering large volumes of therapeutic proteins contained in highly concentrated and / or viscous solutions.

[0274] Combined treatment According to this disclosure, an antibody-drug conjugate containing an anti-CEACAM5 antibody is intended for use in combination with an agent or drug for the treatment of such neuroendocrine cancer. This disclosure also relates to an agent or drug for the treatment of neuroendocrine cancer in combination with an antibody-drug conjugate containing an anti-CEACAM5 antibody.

[0275] In certain embodiments, the subject is pre-treated with an agent or drug for the treatment of neuroendocrine cancers expressing CEACAM5.

[0276] The disclosure also relates to a method for treating cancer in a subject in need, comprising administering an antibody-drug conjugate containing an anti-CEACAM5 antibody to the subject in need, and administering to the subject in need an agent or drug for the treatment of the neuroendocrine cancer.

[0277] This disclosure also relates to agents or drug combinations for the treatment of neuroendocrine cancers, and antibody-drug conjugates comprising an anti-CEACAM5 antibody for the treatment of such cancers.

[0278] As disclosed herein, methods or uses of treatment may achieve improved effects in reducing tumor size.

[0279] As disclosed herein, methods or uses of treatment may achieve improved effects in inhibiting tumor growth.

[0280] This disclosure also relates to agents or drugs for the treatment of such neuroendocrine cancers, and combinations for manufacturing pharmaceuticals for the treatment of neuroendocrine cancers, including antibody-drug conjugates containing an anti-CEACAM5 antibody.

[0281] In one embodiment, this combination enables simultaneous, individual, or sequential administration of an agent or drug for the treatment of neuroendocrine cancer and an antibody-drug conjugate containing an anti-CEACAM5 antibody.

[0282] "Simultaneous administration" refers to the simultaneous administration of both drugs at the same location on the body.

[0283] "Sequential administration" is intended to refer to the administration of both drugs at two consecutive time intervals, at least 30 minutes to several hours, several days, several weeks, or several months, in different or the same body position.

[0284] "Individual administration" is intended to refer to the administration of both drugs at short intervals, i.e., a few seconds, approximately 10-30 seconds, to a few minutes, approximately 1-10 minutes, and at the same or different body positions.

[0285] In one embodiment, this combination enables the simultaneous administration of an agent or drug for the treatment of neuroendocrine cancer and an antibody-drug conjugate containing an anti-CEACAM5 antibody.

[0286] In one embodiment, this combination allows for the individual administration of an agent or drug for the treatment of neuroendocrine cancer and an antibody-drug conjugate containing an anti-CEACAM5 antibody.

[0287] In one embodiment, this combination enables sequential administration of an agent or drug for the treatment of neuroendocrine cancer and an antibody-drug conjugate containing an anti-CEACAM5 antibody.

[0288] In further embodiments, the combination according to this disclosure may be a pharmaceutical composition or a set of components.

[0289] According to some embodiments, antibody-drug conjugates containing anti-CEACAM5 antibodies, and agents or drugs for the treatment of neuroendocrine cancers, are administered simultaneously, individually, or sequentially to the target in need.

[0290] According to one embodiment, an antibody-drug conjugate containing an anti-CEACAM5 antibody and a drug or agent for the treatment of neuroendocrine cancer are administered simultaneously to the target in need. For example, the antibody-drug conjugate containing an anti-CEACAM5 antibody and the drug or agent for the treatment of neuroendocrine cancer are administered approximately simultaneously on day 1 of the cycle.

[0291] According to one embodiment, the antibody-drug conjugate containing an anti-CEACAM5 antibody and the agent or drug for the treatment of neuroendocrine cancer are administered separately to the subject in need. For example, the antibody-drug conjugate containing an anti-CEACAM5 antibody and the agent or drug for the treatment of neuroendocrine cancer are administered on day 1 of the cycle by a different route or to a different location on the subject's body.

[0292] According to one embodiment, an antibody-drug conjugate containing an anti-CEACAM5 antibody and a drug or medication for the treatment of neuroendocrine cancer are administered sequentially to the target in need. For example, the antibody-drug conjugate containing an anti-CEACAM5 antibody and the drug or medication for the treatment of neuroendocrine cancer are administered at different times on day 1 of the cycle.

[0293] In further embodiments, the antibody-drug conjugate containing an anti-CEACAM5 antibody and the agent or drug for the treatment of neuroendocrine cancer may be formulated in the form of (i) a single pharmaceutical composition comprising the antibody-drug conjugate and the agent or drug for the treatment of neuroendocrine cancer, or (ii) two separate pharmaceutical compositions, one of which comprises an antibody-drug conjugate containing an anti-CEACAM5 antibody and the other comprising the agent or drug for the treatment of neuroendocrine cancer. One or more pharmaceutical compositions may also comprise at least one pharmaceutically acceptable excipient.

[0294] When formulation with two separate pharmaceutical compositions, the two separate pharmaceutical compositions may be administered simultaneously, individually, or sequentially to the target that requires them.

[0295] In some embodiments, the pharmaceutical composition is administered sequentially.

[0296] In sequential administration, the time between the administration of the drug or agent for the treatment of neuroendocrine cancer and the administration of the antibody-drug conjugate containing the anti-CEACAM5 antibody may last from about a few minutes to about a few hours, about a few days, or about a few weeks. In some embodiments, the time may range from about 5 minutes to about 3 hours, for example, 10 minutes to about 2.5 hours, about 30 minutes to about 2 hours, or about 1 hour to about 1.5 hours. The time in between may last from about 5 minutes, about 10 minutes, about 30 minutes, about 1 hour, 1.5 hours, about 2 hours, about 2.5 hours, or about 3 hours.

[0297] In some embodiments, in sequential administration on the same day of the cycle, the interval between the administration of the drug or agent for the treatment of neuroendocrine cancer and the administration of the antibody-drug conjugate containing the anti-CEACAM5 antibody may range from about 5 minutes to about 3 hours, for example, 10 minutes to about 2.5 hours, about 30 minutes to about 2 hours, or about 1 hour to about 1.5 hours. In sequential administration on the same day of the cycle, the interval between them may last for about 5 minutes, about 10 minutes, about 30 minutes, about 1 hour, 1.5 hours, about 2 hours, about 2.5 hours, or about 3 hours.

[0298] In sequential administration, antibody-drug conjugates (ADCs) containing anti-CEACAM5 antibodies may be administered before or after the treatment of neuroendocrine cancers or other drugs.

[0299] In some embodiments, an antibody-drug conjugate containing an anti-CEACAM5 antibody may be administered before the agent or drug for the treatment of neuroendocrine cancer.

[0300] In some embodiments, an antibody-drug conjugate containing an anti-CEACAM5 antibody may be administered after an agent or drug for the treatment of neuroendocrine cancer.

[0301] In some embodiments, the order of administration of the antibody-drug conjugate containing the anti-CEACAM5 antibody and the agent or drug for the treatment of neuroendocrine cancer may be the same throughout all treatment cycles.

[0302] In some embodiments, the order of administration of agents or drugs for the treatment of ADCs and neuroendocrine cancers may vary along the treatment cycle. In some embodiments, one or more treatment cycles may include a first administration order, and one or more treatment cycles may include a second administration order, where the first and second orders are different.

[0303] In some embodiments, for example in use for treating cancer, in the first cycle of treatment, the antibody-drug conjugate containing the anti-CEACAM5 antibody may be administered after the agent or drug for treating neuroendocrine cancer, and in subsequent further cycles of treatment, the antibody-drug conjugate containing the anti-CEACAM5 antibody may be administered after the agent or drug for treating neuroendocrine cancer.

[0304] In some embodiments, for example in use for treating cancer, in the first cycle of treatment, the antibody-drug conjugate containing the anti-CEACAM5 antibody may be administered after the agent or drug for treating neuroendocrine cancer, and in subsequent cycles of treatment, the antibody-drug conjugate containing the anti-CEACAM5 antibody may be administered before the agent or drug for treating neuroendocrine cancer.

[0305] In some embodiments, for example in use for treating cancer, the antibody-drug conjugate containing the anti-CEACAM5 antibody may be administered before the agent or drug for treating neuroendocrine cancer in the first cycle of treatment, and in subsequent cycles of treatment, the antibody-drug conjugate containing the anti-CEACAM5 antibody may be administered before the agent or drug for treating neuroendocrine cancer.

[0306] In some embodiments, for example in use for treating cancer, the antibody-drug conjugate containing the anti-CEACAM5 antibody may be administered before the agent or drug for treating neuroendocrine cancer in the first cycle of treatment, and in subsequent cycles of treatment, the antibody-drug conjugate containing the anti-CEACAM5 antibody may be administered after the agent or drug for treating neuroendocrine cancer.

[0307] In some embodiments, for example, in use for treating cancer such as neuroendocrine small cell lung cancer (SCLC), the antibody-drug conjugate containing an anti-CEACAM5 antibody may be administered after the drug or drug for the treatment of neuroendocrine cancer for all cycles of treatment.

[0308] In some embodiments, the agent or drug for treating neuroendocrine cancer may be administered in a loading dose in the first cycle and in a maintenance dose in one or more subsequent cycles.

[0309] In some embodiments, the agent or drug for treating neuroendocrine cancer may be administered in the same dose in the first cycle and one or more subsequent cycles.

[0310] In some embodiments, the antibody-drug conjugate containing an anti-CEACAM5 antibody may be administered at a loading dose in the first cycle and at a maintenance dose in one or more subsequent cycles, and the agent or drug for the treatment of neuroendocrine cancer may be administered at the same dose in the first cycle and one or more subsequent cycles.

[0311] In some embodiments, the antibody-drug conjugate containing an anti-CEACAM5 antibody may be administered in the same dose in the first cycle and one or more subsequent cycles, and the agent or drug for the treatment of neuroendocrine cancer may be administered in the same dose in the first cycle and one or more subsequent cycles.

[0312] In some embodiments, the antibody-drug conjugate and the agent or drug for the treatment of neuroendocrine cancer may be administered on day 1 of the first cycle of treatment and on day 1 of at least one further cycle of treatment.

[0313] Antibody-drug conjugates and agents or drugs for the treatment of neuroendocrine cancers may be administered on day 1 of each treatment cycle.

[0314] The pharmaceutical composition may be one of the above.

[0315] All publications referenced herein are incorporated herein by reference in their entirety for all purposes.

[0316] Sequence List Sequence ID 1: Variable domain of the heavy chain of the anti-CEACAM5 antibody [ka] Sequence ID 2: Variable domain of the light chain of the anti-CEACAM5 antibody DIQMTQSPASLSASVGDRVTITCRASENIFSYLAWYQQKPGKSPKLLVYNTRTLAEGVPSFSGSGSGTDFSLTISSLQPEDFATYYCQHHYGTPFTFGSGTKLEIK Sequence ID 3: HCDR1 anti-CEACAM5 antibody GFVFSSYD Sequence ID 4: HCDR2 of anti-CEACAM5 antibody ISSGGGIT Sequence ID 5: HCDR3 of anti-CEACAM5 antibody AAHYFGSSGPFAY Sequence ID 6: LCDR1 of anti-CEACAM5 antibody ENIFSY Sequence ID 7: LCDR3 of anti-CEACAM5 antibody QHHYGTPFT Sequence ID 8: Heavy chain of anti-CEACAM5 antibody [ka] Sequence ID 9: Light chain of anti-CEACAM5 antibody [ka] [Examples]

[0317] Example 1: Expression of CEACAM5 in neuroendocrine cancer Test group The entire study group consisted of 182 patients, and 183 samples were available (one patient had two different samples available at different points in their clinical course). The selection criteria were: - Diagnosis of neuroendocrine carcinoma, small cell or large cell type, based on morphological and immunohistochemical characteristics (Since there is no consensus on the minimum criteria required to diagnose poorly differentiated neuroendocrine carcinoma, the inventors retained all tumors showing expression of at least one neuroendocrine marker in a significant proportion of tumor cells with suggestive morphologies). - No evidence of mixed tumor, defined by the presence of undifferentiated neuronal components associated with neuroendocrine components. - The tissue material was of sufficient quantity, quality, and quantity for histological studies and immunohistochemical evaluation.

[0318] Materials and methods The following data were recorded: the site of the sample tested, the data of the sample tested, whether the sample was primary or metastatic, and the histological subtype.

[0319] The majority of specimens available for histological and immunohistochemical studies were biopsy specimens. A small number of surgical specimens were also available (it should be noted that surgery is rarely performed for neuroendocrine cancers). A histological review by specialist pathologists was performed on all available tissue material to assess the quantity, quality, and specimen of the tissue samples and to verify the initial diagnosis.

[0320] Immunohistochemical detection of CEACAM5 was performed as per the current protocol (technical and analytical). Detection was carried out using an automated staining system (Ventana Benchmark Ultra, Ventana, Tucson, AZ). 4 μm thick sections of formalin-fixed paraffin-embedded tissue were deparaffinized according to standard procedures. These were incubated in pH 8 demasking solution CC1 (Ventana) at 95°C for 20 minutes, followed by dilution to 1 / 3760 with primary antibody (provided by Sanofi) (final concentration 2.39 μg / mL-1). Development was performed using the ultraViewDAB Universal Detection Kit (Ventana). Counterstaining with hematoxylin was performed. All samples were examined by the same pathologist. All tumor cells with either membranous or cytoplasmic staining were counted as positive, particularly in the small cell type, due to the high proportion of each cytoplasm and the difficulty in accurately positioning the label in these tumor cells where cytoplasm is insufficiently abundant.

[0321] Scoring system used: 0, no positive tumor cells detected; 1, less than 5% positive tumor cells; 2, 5-50% positive tumor cells; 3, 50-80% positive tumor cells; 4, more than 80% positive tumor cells.

[0322] result Characteristics of the test group The study included 183 samples from 182 patients. 133 samples were from primary sites, and 50 samples were from metastatic sites, including liver (n=20), lymph nodes (n=16), peritoneum (n=6), bone (n=3), skin (n=2), and lung (n=1).

[0323] Table 1 shows the distribution of cases based on the location of the primary site.

[0324] [Table 1]

[0325] 111 cases were classified as small cell carcinoma, and 41 cases were classified as large cell carcinoma (mainly from the stomach, colon, pancreas, endometrium, and head and neck). 31 cases were classified as Merkel cell carcinoma and originated from a different subset limited to the skin.

[0326] CEACAM5 expression CEACAM5 tumor cell expression was detected in 53 samples (29%), with scores of 3 or 4 in 20 samples (11%). Interestingly, for patients with two different samples, the score was 0 in one and 1 in the other.

[0327] In the primary tumor site, CEACAM5 expression was detected in 35 samples (26.3%), and the score was 3 or 4 in 12 samples (9%).

[0328] In metastatic sites, CEACAM5 expression in tumor cells was detected in 18 samples (36%), with scores of 3 or 4 in 8 samples (16%).

[0329] Table 2 shows the expression profiles based on the location of the primary site, including both the primary site and the metastatic site.

[0330] [Table 2]

[0331] Tables 3 and 4 show the expression profiles based on the location of the primary site for both the primary site and the metastatic site.

[0332] [Table 3]

[0333] [Table 4]

[0334] Characteristics of CEACAM5+ neuroendocrine cancer There was no significant difference in the number of positive samples between small cell neuroendocrine carcinoma and large cell neuroendocrine carcinoma. Of the 20 samples with a score of 3-4, 16 were from small cell neuroendocrine carcinoma (14.5%) and 4 were from large cell neuroendocrine carcinoma (10%).

[0335] Preliminary conclusion This series of 183 samples is fairly representative of the various locations where neuroendocrine cancers can occur, including some rare or poorly studied sites (female genitalia, head and neck). Some sites are not represented because they were voluntarily excluded (lungs, prostate) or because they are not (or exceptionally not) involved in neuroendocrine cancers (liver, small intestine, appendix) for unknown reasons. Due to the specificity of Gustave Roussy's recruitment, there is some bias in the composition of the series. For example, the absence of a urological surgery department at the institution explains why only metastatic sites were available for the bladder.

[0336] Approximately 30% of the samples contained at least a small number of positive cells. A patient with two different samples available (one showing no positive cells, the other showing less than 5% positive cells) demonstrates the unpredictability of identifying only a small number of positive cells, depending on the extent of the material tested. Scores of 3–4 (50–100% positive cells) may be more reliable, and these were observed in 11% of the samples.

[0337] In positive cases, four locations were concentrated: - Endometrium: 37.5% of positive samples, however, none had scores of 3-4. -Cervix: 60% of positive samples, 13.3% scored 3-4 -Head and neck: 50% of positive samples, 19.2% scored 3-4 -Bladder: 55.5% of positive samples, 11% scored 3-4.

[0338] These results are uniquely interesting because all four locations fall within a rare subset of neuroendocrine cancers, which are consequently understudied and have the least adequately met therapeutic needs.

[0339] Unknown primary neuroendocrine carcinoma is a common clinical problem. This term is used for patients presenting with disseminated disease where a primary site is absent, or, more rarely, for patients diagnosed with primary neuroendocrine carcinoma in a site not known to be involved with the liver or lymph nodes, and without evidence of other tumor masses. In this study, this category is most highlighted in positive cases: 46.2% of positive samples, 31% scored 3-4. This may be clinically relevant.

[0340] Furthermore, the proportion of positive samples was slightly higher at metastatic sites than at primary sites, but this difference was not statistically significant (chi-square test, p>0.15). It is important to re-evaluate the fact that neuroendocrine carcinomas are usually diagnosed at an advanced stage, and that the therapeutic challenge lies in proposing second-line treatments for systemic disease rather than specific treatments targeting late metastases. Possible differences between primary and metastatic sites are therefore not clinically relevant if confirmed by further research.

[0341] In conclusion, this large series indicates that a significant number of neuroendocrine cancer cases, regardless of the primary site, express CEACAM5+ in the majority of tumor cells (approximately 10% of the entire series), and the number of such positive cases is even higher in some specific locations (including the head and neck, female genitalia, and bladder). Screening can be performed by immunohistochemistry, as all patients have at least one diagnostic biopsy, and due to the dissemination of the disease at presentation and its rapid course, repeat biopsies are not a concern here.

[0342] Example 2: Expression of CEACAM5 in medullary thyroid carcinoma the purpose To evaluate CEACAM5 expression in medullary thyroid carcinoma.

[0343] Test sample Sample size: 40

[0344] Origin: Surgical resection (thyroidectomy)

[0345] Stage distribution (known for 31 / 40 samples): 19% Stage 1, 36% Stage 2, 19% Stage 3, 26% Stage 4

[0346] Materials and methods To evaluate the levels and intracellular localization of CEACAM5 protein in the tested samples, immunohistochemistry (IHC) was performed using the mouse parent antibody (769-cea-4) of the antibody tusamitamaburabutansin. IHC was performed using the Ventana Discovery XT automated system with company-recommended reagents (Ventana Medical Systems Inc, USA). Formalin-fixed, paraffin-embedded human tissue slides were dewaxed and pretreated with cell conditioning buffer at 95°C for 48 minutes. The slides were incubated with 769-cea-4 or isotype control mouse IgG1 antibody at a final concentration of 5 μg / mL at 24°C for 3 hours. Rabbit anti-mouse IgG, used as the seed linker antibody, was incubated at a final dilution of 1 / 200 at 24°C for 32 minutes. Ready-to-use biotin-free peroxidase-rich anti-rabbit UltraMap™ was applied at 24°C for 16 minutes. Staining was evaluated under a light microscope, including histological location, main type of reactive cell, staining intensity, and frequency of cell staining.

[0347] result High expression of membrane CEACAM5 was observed in medullary thyroid carcinoma, with a median intensity and incidence of 2+ (60%), and a mean all-red score of 67%.

[0348] The global prevalence was 80% (32 / 40), 35% of the samples expressed CEACAM5 with an intensity of 2 or higher, and the incidence of tumor cells was 50% or higher.

[0349] Example 3: Expression of CEACAM5 in neuroendocrine prostate cancer in patients In this study, the inventors decided to combine morphological analysis and expression of synaptophysin, a common neuroendocrine marker, to evaluate the neuroendocrine differentiation of prostate adenocarcinoma.

[0350] Test group The entire study group consisted of 54 patients. The selection criteria were: -Patients with a known history of prostate adenocarcinoma diagnosed and / or treated at the Gustave Roussy Cancer Campus, including patients participating in the Moscato and MatchR trials. - Pathology reports describing morphologies suggestive of "undifferentiated," "poorly differentiated," or "neuroendocrine" cancer, regardless of the presence or absence of immunohistochemical evidence of neuroendocrine differentiation. - The tissue material was of sufficient quantity, quality, and quantity for histological studies and immunohistochemical evaluation.

[0351] Materials and methods When available, the following data were recorded from clinical and pathological files: date of initial diagnosis of prostate adenocarcinoma, year at initial diagnosis, Gleason grade at initial diagnosis, TNM stage at initial diagnosis, date of diagnosis of neuroendocrine differentiation, site of sample tested, date of sample tested, treatment received, date of last report, and status at last report.

[0352] All specimens available for histological and immunohistochemical testing were biopsy specimens; surgical specimens were unavailable. A histological review was performed on all available tissue material to assess the quantity, quality, and specimen of the tissue samples and to validate the initial diagnosis.

[0353] Further immunohistochemistry was performed. To demonstrate neuroendocrine differentiation of prostate adenocarcinoma, synaptophysin was performed in all cases according to standard laboratory procedures. Detection was performed using an automated staining system (Ventana Benchmark Ultra, Ventana, Tucson, AZ). 4 μm thick sections of formalin-fixed paraffin-embedded tissue were deparaffinized according to standard procedures. They were incubated with primary antibodies (clonal DAK-SYNAP, Dako-Agilent, Santa Clara, CA, ready for use). Development was performed using the ultraViewDAB Universal Detection Kit (Ventana). Counterstaining with hematoxylin was performed. Other specific neuroendocrine markers (chromogranin A, chromogranin B, or INSM1) and CD56 were not performed in all cases at this stage of the study in order to preserve available, and often limited, tissue material. Their expression was recorded when the corresponding tests were performed as part of the initial diagnostic procedure or for other auxiliary studies. In many cases, TTF1 expression, which is expressed in neuroendocrine cancers regardless of origin, was recorded when performed as part of the initial diagnostic procedure.

[0354] Immunohistochemical detection of CEACAM5 was performed according to the current protocol (technical and analytical). Detection was carried out using an automated staining system (Ventana Benchmark Ultra, Ventana, Tucson, AZ). Sections of 4 μm thick formalin-fixed paraffin-embedded tissue were deparaffinized according to standard procedures. These were incubated in pH 8 demasking solution CC1 (Ventana) at 95°C for 20 minutes, followed by dilution to 1 / 3760 with primary antibody (provided by Sanofi) (final concentration 2.39 μg / mL-1). Development was performed using the ultraViewDAB universal detection kit (Ventana). Counterstaining with hematoxylin was performed. All samples were examined by the same pathologist. Multi-head microscopy sessions were conducted with several inventors to evaluate the results and reach a consensus in all cases.

[0355] result Clinical and pathological characteristics of the test group The study included 54 patients. At initial diagnosis, the patients' ages typically fell between 50 and 70 years, with the exception of one patient with a germline BRCA2 mutation, who was diagnosed at age 34. At initial diagnosis, the Gleason grades, where available, were: G6 (n=6), G7 (n=10), G8 (n=6), G9 (n=5), and G10 (n=1).

[0356] Thirty-three patients exhibited immunohistochemical evidence of at least localized neuroendocrine differentiation. Diagnosis of neuroendocrine differentiation was made at the time of diagnosis in one case, or during the course of the disease in 21 cases (after a delay ranging from 1 to 17 years). All cases expressed synaptophysin: 21 cases with over 50% tumor cells, 5 cases with 10-50% tumor cells, and 7 cases with less than 10% tumor cells. Of the 21 cases with over 50% synaptophysin+ tumor cells, 14 cases were tested for chromogranin A (9+, 5-), 2 cases for chromogranin B (2+), and 12 cases for CD56 (10+, 2-). Of the remaining 12 cases, only 4 cases were tested for at least one other neuroendocrine marker. All were negative.

[0357] Twenty-one patients did not show immunohistochemical evidence of neuroendocrine differentiation.

[0358] The site of the biopsy sample used in the test was: -In 11 cases, the primary site or localized dilation was present: prostate (n=5), rectum (n=3), bladder (n=1), pelvis (n=2), - In 43 cases, metastatic sites included: liver (n=20), lymph nodes (18), adrenal gland (n=1), bone (n=3), and peritoneum (n=1).

[0359] CEACAM5 expression CEACAM5 membrane expression was detected in more than 1% of tumor cells in eight cases (range: 1-80%). In two further cases, membrane expression was observed in a small number of scattered tumor cells (less than 1%). Details of the expression pattern (membrane or cytoplasmic location, apparent intensity) for each case are shown in Table 5 below.

[0360] [Table 5]

[0361] [Table 6]

[0362] [Table 7]

[0363] All eight CEACAM5+ cases expressed synaptophysin in over 50% of tumor cells (out of a total of 21 cases with this phenotype). All positive samples were from metastatic sites: six cases (out of 14 cases in this group) from the liver, and two cases (out of 4 cases in this group) from lymph nodes.

[0364] In the remaining 12 cases where synaptophysin was expressed in less than 50% of tumor cells, there was no detectable expression of CEACAM5. In the 21 cases where there was no evidence of synaptophysin expression, there was no expression.

[0365] Preliminary conclusion This study included 54 cases of prostate adenocarcinoma with undifferentiated or poorly differentiated morphologies, of which 33 cases exhibited immunohistochemical evidence of at least focal neuroendocrine differentiation, as assessed by synaptophysin expression. CEACAM5 was detectable only in cases where synaptophysin was expressed in more than 50% of tumor cells. In this patient group, CEACAM5 was detected in more than 1% of tumor cells in 8 out of 21 cases (38%). Therefore, the results suggest that CEACAM5 expression in prostate cancer is associated with the full expression of the neuroendocrine differentiation program, given that synaptophysin is the final product of this program.

[0366] Example 4: Antitumor activity of tusamitamabrabutansin in a PDX model of neuroendocrine prostate cancer Materials and methods Patient-derived xenografts (PDXs) from neuroendocrine pancreatic cancer were selected based on their immunohistochemistry (IHC) and CEACAM5 expression analysis, and transplanted into a mouse model. Patient-derived xenografts (PDXs) from neuroendocrine prostate cancer tumors were selected based on both their immunohistochemical profiling (androgen receptor (AR), synaptophysin (SYP), and CEACAM5) and CEACAM5 mRNA expression (RNA sequencing) analysis.

[0367] Next, five tumors with three phenotypes (2 NEPC CEACAM5+, 1 NEPC CEACAM5 localized, 1 NEPC CEACAM5 and 1 adenocarcinoma) were transplanted into the flanks of 30 NGS mice (NOD.CB17-Prkdcscid / NCrCrl, Charles River). Tumor growth was monitored twice weekly using digital calipers. When the tumor volume reached a mean of 80–200 mm3, the mice were randomized to three different groups: vehicle group (no injection, n=8), unrelated DM4-ADC (35 mg / kg, n=8), and tusamitamabrabutansine (5 mg / kg, n=8). Unrelated ADC or tusamitamabrabutansine was administered intravenously to the tail of the mice. Tumor growth in response to treatment was monitored twice a week, and tumor volume was calculated according to the following formula: L × W × H (L = length, W = width, and H = height). Tumor growth delay was calculated as ΔT / ΔV = (median tumor size T DayY - median tumor size T DayX) / (median tumor size V DayY - median tumor size V DayX) × 100 (where T = treated, V = vehicle, DayY = measurement date, and DayX = start date of treatment). To monitor toxicity, the body weight of mice was measured twice a week. Mice were euthanized if 10% of their initial body weight was lost.

[0368] The average tumor size in the vehicle group was 1500 mm. 3 When the condition was reached, the mice were euthanized. To evaluate circulating CEACAM5, tumors and plasma from the mice were collected at the time of euthanasia.

[0369] Table 6 below shows the expression of CEACAM5 by IHC in different mouse models:

[0370] [Table 8]

[0371] Figure 1 shows the expression levels of CEACAM5 as measured by RNA sequencing.

[0372] To obtain these results, mRNA was extracted from organoids and PDX using the phenol-chloroform method (Sigma, TRI reagent reference 93289). RNA integrity (RNA integrity score ≥ 7.0) was confirmed using an Agilent 2100 Bioanalyzer (Agilent), and the quantity was determined using Qubit (Invitrogen). Following the manufacturer's instructions, the SureSelect Automated Strand Specific RNA Library Preparation Kit was used with the Bravo Platform. Briefly, 50–200 ng of total RNA sample was used for poly-AmRNA selection using oligo(dT) beads, and this was subjected to thermal mRNA fragmentation. The fragmented mRNA sample was subjected to cRNA synthesis and further converted to double-stranded DNA using the reagents supplied in the kit, and the resulting dsDNA was used for library preparation. The final libraries were barcoded, purified, pooled together at equal concentrations, and subjected to paired-end sequencing using a Gustave Roussy Novaseq-6000 sequencer (Illumina). Sequence alignment and gene expression quantification were performed as follows: Raw reads were aligned to human hg38 / GRCh38.p7 and mouse mm10 genomes using STAR. Reads were classified according to their origin species (graft or host) using the XenofilteR tool. The number of reads associated with each gene was obtained using Gencode v26 annotation with STAR (limited to protein-coding genes, antisense, and lincRNAs). Raw counts for each sample were imported into R statistical software. To calculate FPKM (fragment per kilobase transcript per mapped over 1 million fragments) expression levels, the extracted count matrix was normalized to the gene library size and coding length.

[0373] result Tumor contraction Unrelated ADCs used at high doses served as positive controls for the efficacy of chemotherapeutic agents. Indeed, tumor volume was significantly reduced in all models, including those with no or low expression of CEACAM5.

[0374] MR0191 and MR009RO (with high CEACAM5 expression) showed a significant response to a single injection of tusamitamaburabutansine (or SAR408701) compared to the vehicle group at day 21. At day 12, the MR009RO tumor rebounded. This suggested that a second injection at day 12 may improve the response to tusamitamaburabutansine in MR009RO (Figures 2A and 2D).

[0375] MR0084, classified as a localized CEACAM5, showed a significant response, albeit to a lower degree, than that observed in MR191 and MR009RO (Figure 2B).

[0376] The CEACAM5-negative MR059 model and the adenocarcinoma model MR0123 showed no response to tusamitamabrabutansine (Figures 2C and 2E).

[0377] toxicity The fact that body weight changed similarly in vehicle and tusamitamablubutancin-treated mice indicates that no significant toxicity was observed from the treatment (Figures 3A-3E). There was no significant difference in body weight between treated and control mice. Treated mice had similar body weight to control mice and, in particular, much lower tumor burdens in the CEACAM5+ model, indicating that tumor weight was negligible compared to mouse body weight.

[0378] CEACAM5 expression and NEPC marker expression are correlated. Reads were classified according to their origin species (graft or host) using the XenofilteR tool. STAR was used to obtain the number of reads associated with each gene in Gencode v26 annotation (limited to protein-coding genes, antisense, and lincRNAs). The live counts for each sample were imported into R statistical software. The extracted count matrix was normalized to the gene library size and coding length to calculate FPKM (fractures per kilobase transcript per mapped over 1 million fragments) expression levels.

[0379] A significant positive correlation was found between CEACAM5 expression (measured by FPKM from RNA sequencing) and NEPC markers such as SYP, ASCL1, and DLL3 (measured by FPKM from RNA sequencing), while a significant negative correlation was found between CEACAM5 expression and luminal markers such as AR, KLK3, and FOLH1 (Figure 4).

[0380] Example 5: Antitumor activity of tusamitamaburabutansine in patients with neuroendocrine small cell lung cancer (SCLC) (Phase I clinical trial) Main purpose Efficacy was evaluated according to RECIST v1.1 criteria (expansion phase) when SAR408701 was administered once in Q2W, regardless of whether a loading dose was administered in Cycle 1 (expansion phase). The primary efficacy endpoint was a binary response (overall objective response), defined as a confirmed complete response (CR) or partial response (PR) every four cycles using RECIST v1.1.

[0381] statistical design This was a one-cohort, open-label, non-randomized trial in SCLC patients to evaluate the efficacy of tusamitamaburabutansine (SAR408701) administered at a recommended dose of 100 mg / m2 per Q2W cycle in patients with SCLC, in order to establish a preliminary proof-of-concept of antitumor activity in SCLC indications.

[0382] The response rates induced under SAR408701 treatment in this trial were compared to the response rates in historical data with standard treatment. Based on historical data in advanced disease, the response rate for standard treatment was assessed as 10% in the SCLC cohort, and it was expected that if SAR408701 induced a 25% response rate in the SCLC cohort, this would constitute significant antitumor activity. Using this hypothesis, it was assumed that the response rates in the SCLC cohort were 10% under the null hypothesis and 25% under the alternative hypothesis.

[0383] For the SCLC cohort, if at least two responses (confirmed CR (complete response) + PR (partial response)) were observed in the first 25 treated patients, the sample size could be continued to increase up to the total sample size of all 51 treated patients.

[0384] The trial cutoff date was when the last patient in the cohort undergoing treatment had completed two tumor assessments or treatment tumor assessments, regardless of which occurred first to assess antitumor activity.

[0385] Test group For SCLC, recruitment was limited to diseases with a severity of 2+ or higher, involving 1% or more of the tumor cell population, so that CEACAM5 expression in the most recent FFPE-stored tumor tissue samples could be prospectively demonstrated by local or central IHC evaluation.

[0386] The entire study group consisted of 26 patients. The SCLC cohort included patients with CEACAM5 expression of 2+ or higher, representing more than 1% of the tumor cell population.

[0387] The median age of patients was 67.0 years in the range of 49 to 81 years, with 57.7% being 65 years or older. Overall, there were more men than women (65.4% men and 34.6% women), and the majority were Caucasian / Caucasian (73.1%). Most patients (80.8%) were registered with an ECOG score of 1, 15.4% were registered with an ECOG score of 0, and one patient (3.8%) had an ECOG score of 2.

[0388] The selection criteria are, at the discretion of the principal investigator, locally advanced or metastatic solid malignant tumor disease for which standard alternative therapies are unavailable and which meet the following selection criteria: - SCLC patients with CEACAM5 expression in the latest FFPE-stored tumor tissue samples must have an intensity of 2+ or higher, representing 1% or more of the tumor cell population, as prospectively demonstrated by local or central IHC evaluation. - At least 6 × 5 μm slides from formalin-fixed paraffin-embedded (FFPE) stored tissue, and an additional number of slides which may be either 3 × 10 μm (best) or 6 × 5 μm, or equivalent to maintain the same total amount of material required, should be available for local examination at the site and / or shipment to the sponsor or a laboratory designated by the sponsor, evaluation of tumor CEACAM5 expression (retroactively in the escalation phase and predictively in the expansion phase), and exploration of other response predictive biomarkers. If there is insufficient material available, the patient may still be eligible after discussion with the sponsor to assess and confirm that there is enough relevant material for the primary evaluation.

[0389] The selection is enriched for tumors that express or are likely to express CEACAM5, including (but not limited to): - Malignant diseases with a high prevalence of CEACAM5 expression, namely adenocarcinoma or non-sqNSCLC of the large cell subtype, - Circulating cancer-embryonic antigen (CEA) levels > 5 ng / mL, as demonstrated by local testing.

[0390] The recruitment was limited to diseases in which CEACAM5 expression in the latest FFPE-stored tumor tissue samples was 2+ or higher, and included 1% or more of the tumor cell population.

[0391] Of the 26 patients with expanding small cell lung cancer, all experienced metastatic disease, and all had measurable disease. The most common organs involved included the lungs (69.2%), lymph nodes (57.7%), liver (42.3%), brain (34.6%), pleura (26.9%), adrenal glands (23.1%), and bone (19.2%). Slightly more than half of the patients (53.8%) were susceptible, and 46.2% were refractory. CEACAM5 expression at intensity 2+ / 3+ in stored samples was ≥50% in 46.2% of patients (30.8% had 50-80% expression, and 15.4% had ≥80% expression). The median circulating CEA level was 16.00 μg / L, ranging from 0.8 to 5016.3 μg / L. Five patients (21.7%) had circulating CEA levels of ≥100 μg / L.

[0392] CEACAM5 expression levels were essentially recorded retrospectively and centrally in both stored tumor tissue and fresh (baseline) tumor tissue.

[0393] Confirmation of CEACAM5 tumor expression was retrospectively performed in the central laboratory for fresh tumor tissue recovered at baseline. Where sufficient stored tumor material was available, central evaluation was also performed retrospectively to gain insight into the variability of expression assessment. The results of the retrospective analysis did not affect patient management. This was used as a baseline for better interpretation of overall response and for comparison with CEACAM5 expression at disease progression (exploring CEACAM5 loss as a mechanism of acquired resistance).

[0394] The study period for each individual patient included a baseline period of up to 4 weeks, a treatment period of at least one cycle (2 weeks), an end-of-treatment (EOT) evaluation date approximately 30 days after the last administration of the investigational medical product (IMP), and at least one follow-up evaluation date for immunogenicity assessment (approximately 30 days after the EOT evaluation date).

[0395] Pre-treatment All 26 patients with advanced-stage small cell lung cancer had received a chemotherapy regimen prior to the trial. The median number of prior regimens was two (ranging from 1 to 6), and 42.3% of patients had high-dose pretreatment and three or more prior regimens. Most patients (69.2%) had no prior exposure to antitubulin agents. Of the 30.8% of patients who had used antitubulin agents, 87.5% had prior taxane exposure, and 25.0% had prior vinca alkaloid exposure. Most patients with antitubulin exposure had one prior regimen (7 / 8 patients, 87.5%). Most patients (76.9%) had no prior exposure to anti-PD1 / PDL1 agents.

[0396] Investigational drug Tusamitamablubutansine (SAR408701) was administered intravenously as a Q2W cycle, based on actual body surface area, with or without a loading dose in cycle 1, according to the patient cohort in which the patients were enrolled.

[0397] The drug, 100 mg / m² 2 It was administered every two weeks.

[0398] The patient's body surface area (BSA) was calculated using the patient's height and actual weight. 2.2m 2 For patients with BSA exceeding 2.2mg, the dose should be 2.2mg. 2 The calculation was based on BSA.

[0399] Prior medication with a histamine H1 antagonist (diphenylhydramine 50 mg PO or equivalent [e.g., dexchlorpheniramine] administered approximately one hour before tusamitamablubutansine administration) was required for all patients.

[0400] Tusamitamaburabutansine was supplied as a 25 mL extractable volume concentrate for a 125 mg (5 mg / mL) infusion solution contained in a 30 mL Type I glass vial.

[0401] Using an infusion control pump, tusamitamabrabutansine was administered by IV infusion at a rate of 2.5 mg / min for the first 30 minutes, and then increased to 5 mg / min in the absence of hypersensitivity reactions.

[0402] Tusamitamaburabutansine was administered on day 1, and this was repeated every 14 days, with each 14-day period constituting one treatment cycle. Patients could continue treatment until disease progression, unacceptable toxicity occurred, or they wished to discontinue the treatment.

[0403] Since no bacteriostatic agents were present in the product, adherence to sterile techniques was required. Prior to administration, each patient's dose had to be individually prepared by the investigational pharmacist, starting with a pre-filled bag of diluent (0.9% sodium chloride).

[0404] Once the solution was prepared, the dose was administered to the patient within 7.5 hours from bag preparation to completion of dose infusion.

[0405] Two types of administration were used: • Infusion using a syringe driver for low doses (maximum 30 mg / m2). • Pump-assisted infusion for other doses.

[0406] An IV tube administration set fitted with a 0.2-micron filter unit was used for infusion. The test drug was not administered with any other IV fluids. However, the infusion tube was optionally primed with saline or tumami tamaburabutansine. For infusion volumes of 25 mL or less, it was necessary to ensure flushing and elimination of 25 mL of tumami tamaburabutansine before dose infusion. Upon completion of pump infusion, the IV line was flushed with saline as needed to ensure delivery of the entire dose. Upon completion of syringe driver infusion, any remaining tumami tamaburabutansine in the syringe was eliminated.

[0407] safety Of the 26 patients with advanced small cell lung cancer, all experienced at least one TEAE (treatment-related adverse event), with 61.5% experiencing a grade 3 or higher TEAE. Six patients (23.1%) experienced a grade 5 TEAE (all six patients had disease progression: five experienced disease progression, and one had respiratory failure). Fifteen patients (57.7%) experienced a serious TEAE. The majority of patients (80.8%) experienced a TEAE related to the study drug, with 19.2% experiencing a grade 3 or higher related TEAE (none were grade 5). One patient (3.8%) experienced a serious related TEAE (grade 3 or higher AST increase), six patients (23.1%) experienced a TEAE leading to a dose change, and no patients experienced a TEAE leading to permanent discontinuation of treatment.

[0408] In 26 patients with small cell lung cancer in the expanding phase, the most frequently reported SOCs, regardless of the grade of TEAE and its relationship to the investigational procedure, were general and systemic injuries and administration site conditions (61.5%), respiratory, thoracic and mediastinal disorders (53.8%), neurological disorders (50.0%), gastrointestinal disorders (46.2%), metabolic and nutritional disorders (42.3%), and investigation (42.3%).

[0409] Regardless of their association with the study treatment, the most common TEAEs of any grade reported with an incidence of 10% or higher were asthenia (34.6%), dyspnea (30.8%), decreased appetite (26.9%), disease progression (23.1%), elevated AST, constipation, keratitis, nausea (19.2%), cough (15.4%), dizziness, hyponatremia, peripheral edema, peripheral sensory neuropathy, pruritus, and rash (11.5%).

[0410] Patients in the small cell lung cancer cohort did not experience any TEAEs that led to permanent discontinuation of treatment.

[0411] Effectiveness measurement All patients treated during the expansion phase had to have at least one measurable lesion for selection, as efficacy was the primary objective. Tumor assessments were performed at least every four cycles. The decision to continue treatment was based on the investigator's response assessment, but lesion measurements were collected in the e-CRF for the sponsor's response determination. Partial or complete responses had to be confirmed at a second examination at least every four weeks to be recorded as a confirmed response to treatment.

[0412] The primary efficacy endpoint during the expansion phase was overall objective response, defined as confirmed complete response (CR) or partial response (PR) every four cycles using RECIST v1.1. Secondary efficacy variables included duration of response and time to progression (TTP) during the expansion phase.

[0413] A population for which efficacy can be evaluated. The evaluable response population was defined as all treated patients who had a measurable disease at the time of trial enrollment and at at least one evaluable tumor assessment after baseline. Evaluable response patients with small cell lung also had to have a locally documented and criterion-measuring CEACAM5-expressing malignancy with respect to stored tumor tissue.

[0414] Tumor response evaluation To assess objective response or future progression, it was necessary to estimate the overall tumor burden at baseline and use this as a comparison for subsequent measurements. Only patients with measurable disease at baseline were included in protocols where objective tumor response was the primary endpoint. Measurable disease was defined as the presence of at least one measurable lesion. In studies where the primary endpoint was tumor progression (either time to progression or progression as a percentage of a given date), the protocol specified whether participation was limited to individuals with measurable disease or whether patients with only unmeasurable disease were also eligible.

[0415] Efficacy Criteria The criteria for successful response applied are shown in Tables 7 and 8 below.

[0416] [Table 9]

[0417] [Table 10]

[0418] Clear progression of only "non-target" lesions is exceptional, but in such situations, the opinion of the treating physician should take precedence, and the progression should be later confirmed by a review panel (or trial chair).

[0419] Evaluation of the best overall effect Of 26 patients with small cell lung cancer in the expanding stage treated with the recommended dose starting from a dose escalation phase of 100 mg / m2 every two weeks (Q2W), one patient (3.8%) achieved a partial response in cycle 12. The overall response rate (ORR) was 3.8%. Nine patients (34.6%) had the best overall response with stable disease (SD), but one of these patients did not show a partial response (PR) (a decrease of approximately 43.75%).

[0420] Table 9 below provides an overview of the calculation of the overall effect status at each time point for patients with measurable diseases at baseline.

[0421] [Table 11]

[0422] [Table 12]

[0423] If complete regression (CR) is adequately met at the initial point in time, any disease observed at subsequent points in time is considered progressive (PD) at that point, even if it meets the criteria for partial regression (PR) relative to baseline (because the disease is expected to reappear after CR). The best effect depends on whether the minimum duration of stable disease (SD) is matched. However, subsequent scans may still reveal small lesions, and in fact, a "CR" may be claimed if it suggests that the patient had PR rather than CR at the initial point in time. In such circumstances, the original CR should be changed to PR, and the best effect is PR.

[0424] The best overall effect was determined once all the patient data was available.

[0425] Determining the best response in trials where confirmation of complete or partial response is not required: In these trials, the best response was defined as the best response across all time points (for example, a patient with SD at the first evaluation, PR at the second evaluation, and PD at the final evaluation would have the best overall response of PR). If SD was considered the best response, it was also required to meet the protocol specifying the minimum time from baseline. If SD was the best time-point response but did not meet the minimum time, the patient's best response depended on subsequent evaluations. For example, a patient with SD at the first evaluation, PD at the second evaluation, and who did not meet the minimum time for SD would have the best response of PD. The same patient who became unfollowable after the initial SD evaluation would be considered unevaluable.

[0426] Determination of Best Efficacy in Clinical Trials Requiring Confirmation of Complete or Partial Response: Complete or partial response could only be claimed if the criteria for each were met at a subsequent point in time (generally 4 weeks later) as specified in the protocol. In this situation, the best overall response was interpreted as shown in Table 7.

[0427] Duration of response For the 26 patients with advanced small cell lung cancer, the median treatment period was 8.50 weeks, ranging from 2.0 to 52.6 weeks. The median number of cycles administered per patient was 4 cycles, ranging from 1 to 26 cycles. For the only patient in the small cell lung cancer cohort who responded, the duration of response was 6.2 months.

[0428] progress time The median time to tumor progression was 1.9 months (95% CI of 1.68–2.73), and one patient was censored.

[0429] CEACAM5 - Immunohistochemistry (IHC) Scoring Method Immunohistochemical slides stained with CEACAM5 were evaluated by pathologists using a light microscope.

[0430] CEACAM5 positivity was determined by the percentage of surviving tumor cells expressing CEACAM5-positive membrane staining.

[0431] Tumor cells were considered CEACAM5 positive if they showed either partial or complete staining of the surrounding plasma membrane at 2+ or 3+ intensity. If they showed staining at 1+ intensity (weak staining) or no staining (intensity 0), they were considered negative.

[0432] All tumor cells observed in cross-section were evaluated for CEACAM5.

[0433] A minimum of 100 surviving tumor cells were required on the cross-section to determine the proportion of CEACAM5-positive cells.

[0434] The scoring is as follows: %CEACAM5 positivity = 2 + number of tumor cells expressing CEACAM5 membrane staining at or above intensity × 100 / total number of surviving tumor cells present in the cross-section. This captures the percentage of tumor cells stained at each intensity as measured.

[0435] Regarding stored samples, 57.1% of patients expressed CEACAM5 with an intensity of 2 or higher in less than 50% of tumor cells, while 42.9% of patients expressed CEACAM5 with an intensity of 2+ or higher in at least 50% of tumor cells, according to central assessment.

[0436] result CEACAM5 expression Table 11 shows the percentage of CEACAM5-positive cells at an intensity of 2+ or higher, obtained by comparing the results of whole-membrane and polarized film analysis of stored tumor samples. The main pattern of CEACAM5 expression in SCLC was whole-membrane.

[0437] [Table 13]

[0438] Tumor contraction Table 12 shows the best relative tumor shrinkage, stored biopsy, and whole membrane values ​​by class of positive cell percentage at an intensity of 2+ or higher. No association was observed between the best relative tumor shrinkage and CEACAM5 expression at an intensity of 2+ or higher in stored samples (adjusted p-value, 0.2877), however, there was a clear trend towards better tumor shrinkage with higher CEACAM5 expression.

[0439] [Table 14]

[0440] A moderate association was observed between the best relative tumor shrinkage and CEACAM5 expression at a strength of 2+ or higher in stored samples (adjusted p-values ​​of -0.53 and 0.2877, not statistically significant), but there appeared to be a clear trend towards better tumor shrinkage with higher CEACAM5 expression. The prevalence of CEACAM5 at a NB:50% or higher 2+ / 3+ threshold was 11.1%.

[0441] In the SCLC cohort, one confirmed partial response was reported among 26 treated patients. The overall response rate was 3.8% [95%Cl: 0.68–18.89%]. Nine patients reported best overall response (BOR) with stable disease (SD). The duration of response for patients with partial response (PR) was 6.2 months. The median time to progression was 1.9 months.

[0442] conclusion In conclusion, these data demonstrate proof of concept in a subset of neuroendocrine small cell lung cancers treated with tusamitamablubutansine. In particular, these data support the conclusion that tusamitamablubutansine is effective in treating neuroendocrine SCLC. Furthermore, these data support the conclusion that tusamitamablubutansine is especially effective in treating high-CEACAM5-expressing neuroendocrine SCLC.

Claims

1. An immune complex comprising an antibody or an antigen-binding fragment thereof, for use in treating cancers selected from neuroendocrine cancers expressing hCEACAM5 in subjects where such treatment is necessary, wherein the antibody or the antigen-binding fragment thereof specifically binds to hCEACAM5, and the antibody or the antigen-binding fragment thereof is bound to or linked to at least one growth inhibitor, wherein the growth inhibitor is not a topoisomerase I inhibitor.

2. The antibody or the antigen-binding fragment thereof comprises a VH domain and a VL domain, wherein the VH domain comprises three complementarity-determining regions (CDRs) HCDR1, HCDR2, and HCDR3, and the VL domain comprises three CDRs LCDR1, LCDR2, and LCDR3. The HCDR1 is the amino acid sequence of Sequence ID No. 3 【Chemistry 1】 The HCDR2 contains the amino acid sequence of SEQ ID NO:

4. 【Chemistry 2】 The HCDR3 contains the amino acid sequence of SEQ ID NO: 5 【Transformation 3】 Includes, The LDCR1 is the amino acid sequence of Sequence ID No. 6 【Chemistry 4】 The LCDR2 includes the amino acid sequence of NTR, and the LCDR3 includes the amino acid sequence of SEQ ID NO:

7. 【Transformation 5】 including, The immune complex for use according to claim 1.

3. An antibody, or an antigen-binding fragment thereof, or an immune complex comprising the antibody or the antigen-binding fragment thereof, for use in treating cancers selected from neuroendocrine cancers expressing hCEACAM5 in subjects where such treatment is necessary, wherein the antibody or the antigen-binding fragment thereof specifically binds to hCEACAM5, the antibody or the antigen-binding fragment thereof comprises a VH domain and a VL domain, the VH domain comprises the three complementarity-determining regions (CDRs) HCDR1, HCDR2, and HCDR3, and the VL domain comprises the three CDRs LCDR1, LCDR2, and LCDR3. The HCDR1 is the amino acid sequence of Sequence ID No. 3 【Transformation 6】 The HCDR2 contains the amino acid sequence of SEQ ID NO:

4. 【Transformation 7】 The HCDR3 contains the amino acid sequence of SEQ ID NO: 5 【Transformation 8】 The LDCR1 includes the amino acid sequence of SEQ ID NO:

6. 【Chemistry 9】 The LCDR2 includes the amino acid sequence of NTR, and the LCDR3 includes the amino acid sequence of SEQ ID NO:

7. 【Chemistry 10】 Includes, The aforementioned neuroendocrine cancer is not neuroendocrine small cell lung cancer (SCLC). An antibody or its antigen-binding fragment, or an immune complex containing the antibody or its antigen-binding fragment.

4. An antibody, or an antigen-binding fragment thereof, or an immune complex comprising the antibody or the antigen-binding fragment thereof, for use in treating cancers selected from neuroendocrine cancers expressing hCEACAM5 in subjects where such treatment is necessary, wherein the antibody specifically binds to hCEACAM5, and the antibody or the antigen-binding fragment thereof comprises a VH domain and a VL domain, wherein the VH domain comprises the three complementarity-determining regions (CDRs) HCDR1, HCDR2, and HCDR3, and the VL domain comprises the three CDRs LCDR1, LCDR2, and LCDR3. The HCDR1 is the amino acid sequence of Sequence ID No. 3 【Chemistry 11】 The HCDR2 contains the amino acid sequence of SEQ ID NO:

4. 【Chemistry 12】 The HCDR3 contains the amino acid sequence of SEQ ID NO: 5 【Chemistry 13】 The LDCR1 includes the amino acid sequence of SEQ ID NO:

6. 【Chemistry 14】 The LCDR2 includes the amino acid sequence of NTR, and the LCDR3 includes the amino acid sequence of SEQ ID NO:

7. 【Chemistry 15】 including, An antibody, or an antigen-binding fragment thereof, or an immune complex containing the antibody or the antigen-binding fragment thereof.

5. The antibody or immune complex for use according to any one of claims 1 to 4, wherein the cancer is selected from neuroendocrine tumors (NETs), neuroendocrine carcinomas (NECs), mixed neuroendocrine-nonneuroendocrine neoplasms (MiNENs), pheochromocytomas, and medullary thyroid carcinomas (MTCs).

6. The aforementioned cancer, - Neuroendocrine cancers of the gastrointestinal tract and pancreatic and biliary ducts, such as neuroendocrine cancers of the digestive tract, especially the esophagus, stomach, pancreas, liver, small intestine, and large intestine or anal region, or neuroendocrine cancers of the urinary tract, especially neuroendocrine cancers of the bladder. - Neuroendocrine cancers of the upper respiratory tract, gastrointestinal tract, and salivary glands, including neuroendocrine cancers of the lung, especially small cell lung cancer, and neuroendocrine cancers of the thymus. - Neuroendocrine cancers of the thyroid gland, such as medullary thyroid carcinoma (MTC) - Neuroendocrine cancers of the adrenal gland, such as pheochromocytoma, - Neuroendocrine cancers of the skin, such as Merkel cell carcinoma (MCC) - Neuroendocrine cancer of the female reproductive organs such as the endometrium, cervix, or ovaries. - Neuroendocrine cancer of the male reproductive organs, particularly the prostate or testes, and - Neuroendocrine cancer of the head and neck An antibody or immunocomplex for use according to any one of claims 1, 2, and 4, selected from among the following.

7. The antibody or immune complex for use according to any one of claims 1, 2, and 4, wherein the cancer is selected from the neuroendocrine cancers of the esophagus, stomach, small and large intestine, anal region, pancreas, bladder, lung, female reproductive organs, male reproductive organs, thyroid gland, and head and neck.

8. The antibody or immune complex for use according to any one of claims 1, 2, and 4, wherein the cancer is a neuroendocrine cancer of the lung, particularly small cell neuroendocrine lung cancer, or a neuroendocrine cancer of the male reproductive organs, particularly neuroendocrine prostate cancer.

9. The antibody or immune complex for use according to any one of claims 1, 2, and 4 to 8, wherein the cancer is neuroendocrine small cell lung cancer.

10. The antibody or immune complex for use according to any one of claims 1, 2, and 4 to 8, wherein the cancer is neuroendocrine prostate cancer.

11. The antibody or immune complex for use according to any one of claims 1 to 10, wherein the subject is a moderate or high-level expression of carcinoembryonic antigen-associated cell adhesion molecules.

12. The antibody or immune complex for use according to claim 11, wherein the subject is a moderate CEACAM5 expressor, i.e., a subject having CEACAM5 expression of 2+ or 3+ intensity as measured by hCEACAM5 immunohistochemistry in at least 1% of the tumor cell population, or CEACAM5 expression of 1+ intensity in at least 50% of the tumor cell population.

13. The antibody or immune complex for use according to claim 11, wherein the subject is a high CEACAM5 expressor, i.e., a subject having CEACAM5 expression of 2+ or 3+ intensity as measured by hCEACAM5 immunohistochemistry in more than 50% of the tumor cell population.

14. The aforementioned VH domain is sequence number 1: 【Chemistry 16】 An antibody or immunocomplex for use according to any one of claims 2 to 13, comprising:

15. Sequence ID 8: 【Chemistry 17】 An antibody or immunocomplex for use according to any one of claims 1 to 14, comprising a heavy chain containing the above.

16. The aforementioned VL domain is sequence number 2: [Chemistry 18] An antibody or immunocomplex for use according to any one of claims 2 to 15, comprising:

17. Sequence ID 9: 【Chemistry 19】 An antibody or immunocomplex for use according to any one of claims 2 to 16, comprising a light chain containing a light chain.

18. An immune complex comprising the antibody for use according to any one of claims 3 to 17, wherein the antibody is conjugated or linked to at least one growth inhibitor, particularly at least one growth inhibitor which is a cytotoxic agent.

19. The immunocomplex for use according to claim 18, wherein the growth inhibitor is selected from the group consisting of chemotherapeutic agents, enzymes, antibiotics and small molecule toxins or enzymatically active toxins, taxoids, vinca, taxanes, maytansinoids or maytansinoid analogs, tomaimycin or pyrrolobenzodiazepine derivatives, cryptophycin derivatives, leptomycin derivatives, auristatin or drastatin analogs, prodrugs, topoisomerase inhibitors, DNA alkylating agents, antitubulin agents and toxins such as CC-1065 or CC-1065 analogs.

20. The immunocomplex for use according to claim 18, wherein the growth inhibitor is not a topoisomerase I inhibitor.

21. The aforementioned proliferation inhibitor is (N ２ '-Deacetyl-N ２ '-(3-mercapto-1-oxopropyl)-meitansine) (DM1) or N ２ '-Deacetyl-N ２ An immune complex for use according to any one of claims 1 to 2 and 5 to 20, wherein the immune complex is '-(4-methyl-4-mercapto-1-oxopentyl)-meytansine (DM4).

22. An immunocomplex for use according to any one of claims 1 to 2 and 5 to 21, wherein the antibody is covalently bound to at least one growth inhibitor via a cleavable or non-cleavable linker, particularly via a linker selected from the group consisting of N-succinimidylpyridiyl dithiobutyrate (SPDB), 4-(pyridine-2-yldisulfanyl)-2-sulfobutyrate (sulfo-SPDB), and succinimidyl (N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC).

23. It comprises a heavy chain consisting of SEQ ID NO: 8 and a light chain consisting of SEQ ID NO: 9, and is transmitted via N-succinimidylpyridiyl dithiobthiolate (SPDB), ２ '-Deacetyl-N ２ An immunocomplex for use according to any one of claims 1 to 22, comprising an hCEACAM5 antibody covalently bound to (4-methyl-4-mercapto-1-oxopentyl)-meytansine (DM4).

24. The immune complex for use according to any one of claims 1 to 23, wherein the immune complex is tusamitamabrabutansin.

25. The antibody or immune complex is administered in a concentration of 5, 10, 20, 30, 40, 60, 80, 100, 120, 150, 180, or 210 mg / m², based on the target body surface area. ２ An antibody or immune complex for use according to any one of claims 1 to 24, administered at a dose level.

26. The antibody or immune complex for use according to any one of claims 1 to 25, wherein the antibody or immune complex is administered every two weeks or every three weeks.

27. The antibody or immune complex is administered every two weeks at a dose of 100 mg / m² based on the surface area of ​​the target body. ２ An antibody or immune complex for use according to any one of claims 1 to 26, administered at a dose level of [specified dose level].

28. The antibody or the immune complex is administered as a loading dose of about 100 mg / m ２ to about 200 mg / m ２ , particularly as a loading dose, about 135 mg / m ２ , about 150 mg / m ２ , or 170 mg / m ２ The antibody or immune complex for use according to any one of claims 1 to 27, which is administered at a dose of.

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