WO2024251733A1 - Antitumor combinations containing anti-ceacam5 antibody-drug conjugates, anti-pd1/pd-l1 antibodies and anti-ctla4 antibodies - Google Patents

Abstract

The disclosure relates to a combination of (i) an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody, (ii) an anti-PD-1 antibody or anti-PD-L1 antibody, and (ii) an anti-CTLA4 antibody, for use in the treatment of a cancer.

Description

[TITLE]

ANTITUMOR COMBINATIONS CONTAINING ANTI-CEACAM5 ANTIBODY-DRUG CONJUGATES, ANTI-PD1/PD-L1 ANTIBODIES AND ANTI-CTLA4 ANTIBODIES

[REFERENCE TO SEQUENCE LISTING]

[0001] The instant application contains a Sequence Listing which has been submitted electronically in. xml format (ST26 WIPO STD) and is hereby incorporated by reference in its entirety. Said .xml copy, created on June 1 , 2023, is named PR94703_EP_SANOFI_LISTING SEQUENCES.xml.

[TECHNICAL FIELD]

[0002] The present disclosure relates to the field of therapeutic treatment of cancers that express CEACAM5. Certain aspects of the disclosure relate to combination therapies with an immunoconjugate comprising an anti-CEACAM5 antibody, with an anti-CTLA4 antibody, and with an anti-PD-1 or anti-PD-L1 agent, to treat cancer, including lung, gastric, gastroesophageal junction, esophageal, colorectal and pancreatic cancers.

[TECHNICAL BACKGROUND]

[0003] According to the World Health Organization, cancer was the second leading cause of death globally and responsible for approx. 9.6 million in 2018. Thus, there is a continued need for providing improved drug combinations and regimens for the treatment of cancer.

[0004] Carcino-embryonic antigen (CEA) is a glycoprotein involved in cell adhesion. CEA was first identified in 1965 (Gold and Freedman, J Exp Med, 121 , 439, 1965) as a protein normally expressed by fetal gut during the first six months of gestation, and found in cancers of the pancreas, liver, and colon. The CEA family belongs to the immunoglobulin superfamily. The CEA family, which consists of 18 genes, is sub-divided in two sub-groups of proteins: the carcinoembryonic antigen-related cell adhesion molecule (CEACAM) sub-group and the pregnancy-specific glycoprotein subgroup (Kammerer & Zimmermann, BMC Biology 2010, 8:12).

[0005] In humans, the CEACAM sub-group consists of 7 members: CEACAM1 , CEACAM3, CEACAM4, CEACAM5, CEACAM6, CEACAM7 and CEACAM8. Numerous studies have shown that CEACAM5, identical to the originally identified CEA, is highly expressed on the surface of colorectal, gastric, lung, breast, prostate, ovary, cervix, and bladder tumor cells and weakly expressed in few normal epithelial tissues such as columnar epithelial and goblet cells in colon, mucous neck cells in the stomach and squamous epithelial cells in esophagus and cervix (Hammarstrom et al, 2002, in “Tumor markers, Physiology, Pathobiology, Technology and Clinical Applications” Eds. Diamandis E. P. et al., AACC Press, Washington pp 375). Thus, CEACAM5 may constitute a therapeutic target suitable for tumor specific targeting approaches, such as antibody-drug conjugates (ADCs).

[0006] Antibody-drug conjugates (ADCs) comprise an antibody attached to a chemotherapeutic agent such as a cytotoxic agent or a growth inhibitory agent or a cytostatic agent. The chemotherapeutic agent is typically attached to the antibody via a chemical linker. These antibody-drug conjugates (ADCs) have great potential in cancer chemotherapy and enable selective delivery of a potent chemotherapeutic agent to target cancer cells, resulting in improved efficacy, reduced systemic toxicity, and improved pharmacokinetics, pharmacodynamics and biodistribution compared to traditional chemotherapy. To date, hundreds of diverse antibody-drug conjugates (ADCs) have been developed against various cancers, of which several have been approved for human use.

[0007] The mechanism of action of antibody drug conjugates (ADCs) begins with its binding to a specific antigen, sufficiently expressed on the tumor cells in order to achieve a selective and efficient internalization of the drug. Selectively targeting potent cytotoxic agents to tumor cells using ADCs has now been shown to be an effective strategy for the treatment of cancer, as demonstrated by the recent approvals of brentuximab vedotin for the treatment of Hodgkin lymphoma and trastuzumab emtansine (T-DM1 ) for the treatment of relapsed metastatic HER2+ breast cancer (Younes A, Gopal AK, Smith SE, Ansell SM, Rosenblatt JD, Savage KJ, et al. Results of a pivotal phase II study of brentuximab vedotin for patients with relapsed or refractory Hodgkin's lymphoma. J Clin Oncol. 2012;30(18) :2183-9; Verma S, Miles D, Gianni L, Krop IE, Welslau M, Baselga J, et al. Trastuzumab emtansine for HER2-positive advanced breast cancer. N Engl J Med. 2012;19:1783-91 ). Many other malignant diseases with unmet medical needs, such as solid tumor cancers, could benefit from such therapeutic options.

[0008] The international patent application published as WO 2014/079886 discloses an antibody binding to the A3-B3 domain of human and Macaca fascicularis CEACAM5 proteins and which does not significantly cross-react with human CEACAM1 , human CEACAM6, human CEACAM7, human CEACAM8, Macaca fascicularis CEACAM1 , Macaca fascicularis CEACAM6, and Macaca fascicularis CEACAM8. This antibody has been conjugated to a maytansinoid, thereby providing an antibody-drug conjugate having a significant cytotoxic activity on MKN45 human gastric cancer cells, with IC50 values < 1 nM.

[0009] huMAb2-3-SPDB-DM4 disclosed in WO 2014/079886 is an immunoconjugate (antibody-drug conjugate, ADC) comprising a humanized anti-CEACAM5 antibody linked to maytansinoid derivative 4 (DM4), a potent antimitotic agent that inhibits microtubule assembly. DM4 is covalently bound to the antibody through an optimized linker SPDB [N-succinimidyl-4- (2-pyridyldithio)butanoic acid] that is stable in plasma and cleavable inside cells. After binding and internalization in targeted cancer cells, huMAb2-3-SPDB-DM4 is degraded, releasing cytotoxic DM4 metabolites. huMAb2-3-SPDB-DM4 is also known as tusamitamab ravtansine.

[0010] WO 2020/161214, the entire content of which is incorporated herein by reference, discloses use of anti-CEACAM5 immunoconjugates (ADCs) for treating lung cancer.

[0011] Immune checkpoint inhibitors (ICIs), administered alone or in combination with other anticancer therapies, have shown activity in treating a variety of tumor types, including non-small-cell lung cancer (NSCLC), with a manageable safety profile when combined with cytotoxic agents.

[0012] The cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) is an inhibitory regulator of T-cell mediated responses. It has been studied as a target of monoclonal antibodies for cancer immunotherapy. CTLA-4 was the first immune checkpoint to be targeted by an Immune checkpoint inhibitor (ICI), with the all-human immunoglobulin G1 (lgG1 ) monoclonal anti-CTLA-4 antibody ipilimumab being approved by the FDA in 2011 .

[0013] Over the past decade, cancer therapies involving CTLA-4 inhibitor monotherapy have been largely replaced by combination therapies combining CTLA-4 inhibitors with anti- PD-1/PD-L1 antibodies. Anti-CTLA-4 antibodies have been shown to enhance the efficacy of PD-1/PD-L1 blockade in various solid tumors, with additive or even synergistic effects.

[0014] Programmed death 1 (PD-1 ) is a cell surface receptor acting as a T cell checkpoint and regulating T cell depletion. Binding of PD-1 T cell activation. Abnormal high expression of PD-L1 on tumor cells and antigen-presenting cells in the tumor microenvironment mediates tumor immune escape. The development of anti-PD-1/PD-L1 antibodies has recently attracted a lot of interest in cancer immunotherapy.

[0015] Pembrolizumab (KEYTRUDA®), a humanized lgG4 monoclonal antibody against programmed cell death protein 1 (PD-1 ), is indicated as a single agent as the first-line treatment of patients with NSCLC expressing programmed death-ligand 1 protein (PD-L1 ). [0016] Despite recent progress in the treatment of cancer such as advanced NSCLC, there remains a need for effective new treatment at the time of disease progression.

[0017] The present disclosure has for object to satisfy all or part of this need.

[SUMMARY]

[0018] According to one of its objects, the present disclosure relates to a combination of (i) an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody and (ii) an anti-CTLA4 antibody, for use in the treatment of a cancer.

[0019] In some embodiments, the present disclosure relates to an antibody-drug conjugate (ADC) comprising an anti-CEACAM5-antibody for use in combination with an anti- CTLA4 antibody for the treatment of cancer.

[0020] The present disclosure relates to an anti-CTLA4 antibody for use in combination with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5-antibody and a chemotherapeutic agent for the treatment of cancer

[0021] In the disclosure, “antibody-drug conjugate comprising an anti-CEACAM5- antibody”, the “antibody-drug conjugate” and “ADC” are used interchangeably.

[0022] In some embodiments, the ADC and the anti-CTLA4 antibody may be each in an effective amount.

[0023] In some embodiments, the use may be in a subject in need thereof.

[0024] In some embodiments, the cancer may be a CEACAM5-positive cancer, i.e., the cancer (or tumor) cells express CEACAM5.

[0025] As shown in the Examples section, it has been observed that the administration of a combination comprising an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody, such as tusamitamab ravtansine (huMAb2-3-SPDB-DM4), an anti-CTLA4 antibody, such as durvalumab and tremelimumab, and an anti-PD1 antibody was highly efficient for reducing or preventing the tumor growth, and even for reducing tumor size, in various syngeneic colorectal mouse models.

[0026] Compared to the ADC alone and to the combination anti-CTLA4/anti-PD1 antibodies, the combination ADC + anti-CTLA4/anti-PD1 antibodies was able to induce partial and complete response in about 50 to about 100% of the treated mice. Further, the combination ADC + anti-CTLA4/anti-PD1 antibodies was able to reduce AT/AC below 10%, or even below 0%, revealing a synergistic effect of the combination compared to the ADC alone and to the combination anti-CTLA4/anti-PD1 antibodies.

[0027] Further, the results show that a synergistic effect was obtained with the ADC, anti-CTLA4 antibody and anti-PD1/PDL1 antibody each used at a sub-optimal dose. Advantageously, the obtaining of a synergistic effect at suboptimal doses allows maintaining a good therapeutic effect while reducing the risk of an adverse event.

[0028] Also, it was surprisingly observed that the combination of ADC + anti-CTLA4 antibody was able to induce a partial or complete regression in about 30% of the treated mice, while no regression at all could be observed in mice treated with the ADC alone of with the anti-CTLA4 antibody alone. Also, the combination of ADC + anti-CTLA4 antibody was able to reduce AT/AC below 10%, revealing a synergistic effect of the combination compared to the ADC alone and the anti-CTLA4 antibody alone.

[0029] In some embodiments, the anti-CEACAM5 antibody may comprise a CDR-H1 having the amino acid sequence of SEQ ID NO: 1 , a CDR-H2 having the amino acid sequence of SEQ ID NO: 2, a CDR-H3 having the amino acid sequence of SEQ ID NO: 3, a CDR-L1 having the amino acid sequence of SEQ ID NO: 4, a CDR-L2 having the amino acid sequence NTR, and a CDR-L3 having the amino acid sequence of SEQ ID NO: 5.

[0030] In some embodiments, the anti-CEACAM5 antibody may comprise a variable domain of a heavy chain (VH) consisting of SEQ ID NO: 6, or a sequence at least 85%, or at least 90% identical thereto and comprising the CDR-H1 , CDR-H2 and CDR-H3 as above indicated, and a variable domain of a light chain (VL) consisting of SEQ ID NO: 7, or a sequence at least 85%, or at least 90% identical thereto and comprising the CDR-L1 , CDR- L2 and CDR-L3 as above indicated.

[0031] In some embodiments, the anti-CEACAM5 antibody may comprise a variable domain of a heavy chain (VH) consisting of SEQ ID NO: 6 and a variable domain of a light chain (VL) consisting of SEQ ID NO: 7.

[0032] In some embodiments, the anti-CEACAM5 antibody may be tusamitamab.

[0033] In some embodiments, the antibody-drug conjugate may comprise at least one chemotherapeutic agent.

[0034] In some embodiments, the chemotherapeutic agent may be selected from the group consisting of radioisotopes, protein toxins, small molecule toxins, and any combination thereof. [0035] In some embodiments, the small molecule toxin may be selected from antimetabolites, DNA-alkylating agents, DNA-cross-linking agents, DNA-intercalating agents, anti-microtubule agents, topoisomerase inhibitors, and any combination thereof.

[0036] In some embodiments, the anti-microtubule agent may be selected from taxanes, vinca alkaloids, maytansinoids, colchicine, podophyllotoxin, griseofulvin, and any combination thereof.

[0037] In some embodiments, the chemotherapeutic agent may be a maytansinoid.

[0038] In some embodiments, the maytansinoid may be selected from N2’-deacetyl- N2’-(3-mercapto-1 -oxopropyl)-maytansine (DM1 ), N2’-deacetyl-N2’-(4-methyl-4-mercapto-1 - oxopentyl)-maytansine (DM4), and combinations thereof.

[0039] In some embodiments, the anti-CEACAM5 antibody may be covalently attached via a cleavable or non-cleavable linker to the at least one chemotherapeutic agent.

[0040] In some embodiments, linker may be selected from N-succinimidyl pyridyldithiobutyrate (SPDB), 4-(pyridin-2-yldisulfanyl)-2-sulfo-butyric acid (sulfo-SPDB), and succinimidyl(N-maleimidomethyl) cyclohexane-1 -carboxylate (SMCC).

[0041 ] In some embodiments, the anti-CEACAM5 antibody may be covalently attached via a cleavable linker to the at least one chemotherapeutic agent, the linker being selected from N-succinimidyl pyridyldithiobutyrate (SPDB), 4-(pyridin-2-yldisulfanyl)-2-sulfo-butyric acid (sulfo-SPDB), and succinimidyl(N-maleimidomethyl) cyclohexane-1 -carboxylate (SMCC).

[0042] In some embodiments, the CEACAM5-antibody may comprise a heavy chain (HC) consisting of SEQ ID NO: 8, or a sequence at least 85%, or at least 90% identical thereto and comprising the CDR-H1 , CDR-H2 and CDR-H3 as above indicated, and a light chain (LC) consisting of SEQ ID NO: 9, or a sequence at least 85%, or at least 90% identical thereto and comprising the CDR-L1 , CDR-L2 and CDR-L3 as above indicated.

[0043] In some embodiments, the CEACAM5-antibody may comprise a heavy chain (HC) consisting of SEQ ID NO: 8 and a light chain (LC) consisting of SEQ ID NO: 9 (huMAb2- 3).

[0044] In some embodiments, the CEACAM5-antibody may comprise a heavy chain (HC) consisting of SEQ ID NO: 8 and a light chain (LC) consisting of SEQ ID NO: 9 and which may be covalently linked to N2’-deacetyl-N-2’(4-methyl-4-mercapto-1 -oxopentyl)-maytansine (DM4) via N-succinimidyl pyridyldithiobutyrate (SPDB).

[0045] In some embodiments, the antibody-drug conjugate may be characterized by a drug-to-antibody ratio (DAR) ranging from 1 to 10. [0046] In some embodiments, the antibody-drug conjugate may be tusamitamab ravtansine.

[0047] In some embodiments, the antibody-drug conjugate may be administered at a dose of 80 mg/m² to 210 mg/m², 80 mg/m² to 170 mg/m², or at a dose of 80 mg/m² to 150 mg/m², or at a dose of 80 mg/m² to 120 mg/m², or at a dose of 80 mg/m² to 100 mg/m².

[0048] In some embodiments, the antibody-drug conjugate may be administered at a dose level of 80, 100, 120, 150, 170, 180, or 210 mg/m².

[0049] In some embodiments, the CTLA4 antibody may be ipilimumab, tremelimumab, quavonlimab, durvalumab, or zalifrelimab.

[0050] In some embodiments, the CTLA4 antibody may be tremelimumab or durvalumab.

[0051] In some embodiments, the CTLA4 antibody may be tremelimumab.

[0052] In some embodiments, the CTLA4 antibody may be durvalumab.

[0053] In some embodiments, the CTLA4 antibody may be administered at a dose of 1 mg/kg to 10 mg/kg, or at a dose of 3 mg/kg to 10 mg/kg.

[0054] In some embodiments, the CTLA4 antibody may be administered at a dose 1 mg/kg, or at a dose of 2.5 mg/kg, or at a dose of 3 mg/kg, or at a dose of 4 mg/kg, or at a dose of 10 mg/kg.

[0055] In some embodiments, the combination may be administered to a subject having not received any prior systemic chemotherapy for treatment of the cancer.

[0056] In some embodiments, (i) the antibody-drug conjugate and (ii) the anti-CTLA4 antibody may be administered on day 1 of a first cycle of treatment. In some embodiments, (i) the antibody-drug conjugate and (ii) the anti-CTLA4 antibody may be administered on day 1 of a first cycle of treatment and on day 1 of at least one additional cycle(s) of treatment.

[0057] In some embodiments, (i) the antibody-drug conjugate may be administered before or after the (ii) the anti-CTLA4 antibody.

[0058] In some embodiments, (i) the antibody-drug conjugate may be administered before the (ii) the anti-CTLA4 antibody. [0059] In some embodiments, (i) the antibody-drug conjugate may be administered on day 1 of a first cycle of treatment and (ii) the anti-CTLA4 antibody may be administered at a subsequent day of the cycle of treatment. In some embodiments, (i) the antibody-drug conjugate may be administered on day 1 of a first cycle of treatment and on day 1 of at least one additional cycle(s) of treatment, and (ii) the anti-CTLA4 antibody may be administered at a subsequent day of treatment of the cycle of treatment and at a subsequent day of the at least one additional cycle(s) of treatment.

[0060] In some embodiments, a cycle of treatment may be from 1 to 4 weeks, or may be 1 , 2, 3 or 4 weeks, or may be 7, 10, 13, 14, 17, 20, or 21 days.

[0061] In some embodiments, (i) the antibody-drug conjugate and (ii) the anti-CTLA4 antibody may be administered once every three weeks or once every two weeks.

[0062] In some embodiments, (i) the antibody-drug conjugate and (ii) the anti-CTLA4 antibody may be simultaneously, separately, or sequentially administered.

[0063] In some embodiments, (i) the antibody-drug conjugate and (ii) the anti-CTLA4 antibody may be sequentially administered.

[0064] In some embodiments, the combination may further comprise (iii) an anti-PD-1 antibody or anti-PD-L1 antibody.

[0065] In some embodiments, the anti-PD-1 antibody may be selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, sintilimab, dostarlimab, and tislelizumab.

[0066] In some embodiments, the anti-PD-1 antibody may be pembrolizumab or sintilimab.

[0067] In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of 150 mg to 400 mg, or at a dose of 150 mg to 300 mg.

[0068] In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of 200 mg.

[0069] In some embodiments, (i) the antibody-drug conjugate, (ii) the anti-CTLA4 antibody, and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered on day 1 of a first cycle of treatment. In some embodiments, (i) the antibody-drug conjugate, (ii) the anti-CTLA4 antibody, and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered on day 1 of a first cycle of treatment and on day 1 of at least one additional cycle(s) of treatment. [0070] In some embodiments, (i) the antibody-drug conjugate may be administered before or after (ii) the anti-CTLA4 antibody and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody.

[0071] In some embodiments, (i) the antibody-drug conjugate may be administered before (ii) the anti-CTLA4 antibody and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody.

[0072] In some embodiments, (i) the antibody-drug conjugate may be administered on day 1 of a first cycle of treatment and (ii) the anti-CTLA4 antibody and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a subsequent day of the cycle of treatment. In some embodiments, (i) the antibody-drug conjugate may be administered on day 1 of a first cycle of treatment and on day 1 of at least one additional cycle(s) of treatment, and (ii) the anti-CTLA4 antibody and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a subsequent day of treatment of the cycle of treatment and at a subsequent day of the at least one additional cycle(s) of treatment

[0073] In some embodiments, a cycle of treatment may be from 1 to 4 weeks, or may be 1 , 2, 3 or 4 weeks, or may be 7, 10, 13, 14, 17, 20, or 21 days.

[0074] In some embodiments, (i) the antibody-drug conjugate, (ii) the anti-CTLA4 antibody, and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered once every three weeks or once every two weeks.

[0075] In some embodiments, (i) the antibody-drug conjugate, (ii) the anti-CTLA4 antibody, and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody may be simultaneously, separately, or sequentially administered.

[0076] In some embodiments, (i) the antibody-drug conjugate, (ii) the anti-CTLA4 antibody, and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody may be sequentially administered.

[0077] In some embodiments, (ii) the anti-CTLA4 antibody and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody may be simultaneously administered.

[0078] In some embodiments, (i) the antibody-drug conjugate may be administered at a dose of 80 mg/m² to 210 mg/m², 80 mg/m² to 170 mg/m², or at a dose of 80 mg/m² to 150 mg/m², or at a dose of 80 mg/m² to 120 mg/m², or at a dose of 80 mg/m² to 100 mg/m², (ii) the anti-CTLA4 antibody may be administered at a dose of 1 mg/kg to 10 mg/kg, or at a dose of 3 mg/k to 10 mg/kg, and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of 150 mg to 400 mg, or at a dose of 150 mg to 300 mg. [0079] In some embodiments, (i) the antibody-drug conjugate may be administered at a dose level of 80, 100, 120, 150, 170, 180, or 210 mg/m² (ii) the anti-CTLA4 antibody may be administered at a dose of 1 mg/kg, or at a dose of 2.5 mg/kg, or at a dose of 3 mg/kg, or at a dose of 4 mg/kg, or at a dose of 10 mg/kg, and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of 200 mg.

[0080] In some embodiments, the anti-PD1/PDL1 antibody and the anti-CTLA4 antibody may be provided as a bispecific antibody directed to PD1/PDL1 and CTLA4.

[0081] In some embodiments, the cancer may be a CEACAM5 positive cancer.

[0082] In some embodiments, the cancer expresses CEACAM5 with moderate or high intensity defined by immunohistochemistry.

[0083] In some embodiments, the cancer may be a CEACAM5 positive cancer having a CEACAM5 immunohistochemical intensity > 2+ in > 1 % and < 50% of cancer cells. Such cancer expresses CEACAM5 with moderate intensity.

[0084] In some embodiments, the cancer may be a CEACAM5 positive cancer having a CEACAM5 immunohistochemical intensity > 2+ in > 50% of cancer cells. Such cancer expresses CEACAM5 with high intensity.

[0085] In some embodiments, the cancer may be selected from hepatocellular carcinoma, colorectal cancer, gastric cancer, gastroesophageal junction adenocarcinoma (GEJ), esophageal cancer, lung cancer, uterine cervix cancer, pancreatic cancer, ovarian cancer, thyroid cancer, bladder cancer, endometrial cancer, breast cancer, liver cancer, biliary tract cancer (e.g., cholangiocarcinoma), prostate cancer, neuroendocrine cancer, and skin cancer.

[0086] In some embodiments, the cancer may be selected from a colorectal cancer, a gastric cancer, a gastroesophageal junction adenocarcinoma (GEJ), an esophageal cancer, a pancreatic cancer and a lung cancer.

[0087] In some embodiments, the cancer may be a colorectal cancer.

[0088] In some embodiments, the cancer may be a pancreatic cancer.

[0089] In some embodiments, the cancer may be a gastric cancer, a gastroesophageal junction (GEJ) adenocarcinoma, or an esophageal cancer.

[0090] In some embodiments, the cancer may be a lung cancer. [0091] In some embodiments, the lung cancer may be a non-squamous non-small cell lung cancer (NSQ NSCLC).

[0092] In some embodiments, the non-squamous non-small cell lung cancer may be an advanced or metastatic NSQ NSCLC.

[0093] In some embodiments, the non-squamous non-small cell lung cancer has no epidermal growth factor receptor (EGFR) sensitizing mutation or v-raf murine sarcoma viral oncogene homolog B1 (BRAF) mutation or anaplastic lymphoma kinase/c-ros oncogene 1 (ALK/ROS) alterations.

[0094] According to one of its objects, the present disclosure relates to a pharmaceutical composition comprising (i) an antibody-drug conjugate as disclosed herein, (ii) an anti-CTLA4 antibody, and a pharmaceutically acceptable excipient.

[0095] According to one of its objects, the present disclosure relates to a pharmaceutical composition comprising (i) an antibody-drug conjugate as disclosed herein, (ii) an anti-CTLA4 antibody, and (iii) an anti-PD-1 antibody or an anti-PD-L1 antibody, and a pharmaceutically acceptable excipient.

[0096] In some embodiments, (i) the antibody-drug conjugate and (ii) the anti-CTLA4 antibody may be formulated in the form of two separate pharmaceutical compositions, wherein (a) a first pharmaceutical composition may comprise the antibody-drug conjugate and a pharmaceutically acceptable excipient, and (b) a second pharmaceutical composition may comprise the anti-CTLA4 antibody and a pharmaceutically acceptable excipient.

[0097] In some embodiments, (i) the antibody-drug conjugate and (ii) the anti-CTLA4 antibody, and (iii) an anti-PD-1 antibody or an anti-PD-L1 antibody may be formulated in the form of three separate pharmaceutical compositions, wherein (a) a first pharmaceutical composition may comprise the antibody-drug conjugate and a pharmaceutically acceptable excipient, (b) a second pharmaceutical composition may comprise the anti-CTLA4 antibody and a pharmaceutically acceptable excipient, and c) a third pharmaceutical composition may comprise the anti-PD-1 antibody or the anti-PD-L1 antibody and a pharmaceutically acceptable excipient.

[0098] According to one of its objects, the present disclosure relates to a kit-of-parts comprising (i) a pharmaceutical composition comprising an antibody-drug conjugate as disclosed herein, and a pharmaceutically acceptable excipient, and (ii) a pharmaceutical composition comprising an anti-CTLA4 antibody and a pharmaceutically acceptable excipient, and, optionally, (ill) a pharmaceutical composition comprising an anti-PD-1 antibody or an anti- PD-L1 antibody and a pharmaceutically acceptable excipient.

[0099] According to one of its objects, the present disclosure relates to a pharmaceutical composition as disclosed herein, or a kit as disclosed herein, for use in the treatment of a cancer.

[0100] According to one of its objects, the present disclosure relates to a use of a combination of (i) an antibody-drug conjugate as disclosed herein and (ii) an anti-CTLA4 antibody for the manufacture of a pharmaceutical composition or a kit for the treatment of a cancer.

[0101] According to one of its objects, the present disclosure relates to a use of a combination of (i) an antibody-drug conjugate as disclosed herein, (ii) an anti-CTLA4 antibody, and (iii) an anti-PD-1 antibody or anti-PD-L1 antibody for the manufacture of a pharmaceutical composition or a kit, for the treatment of a cancer.

[0102] According to one of its objects, the present disclosure relates to a method for treating a cancer in a subject in need thereof, the method comprising at least a step of administering a combination of (i) an antibody-drug conjugate as disclosed herein and (ii) an anti-CTLA4 antibody.

[0103] According to one of its objects, the present disclosure relates to a method for treating a cancer in a subject in need thereof, the method comprising at least a step of administering a combination of (i) an antibody-drug conjugate as disclosed herein, (ii) an anti- CTLA4 antibody, and (iii) an anti-PD-1 antibody or anti-PD-L1 antibody.

[0104] In the methods of the disclosure, the administration may be simultaneous, separate or sequential.

[0105] In the methods of the disclosure, the administration of (i) an antibody-drug conjugate as disclosed herein may be sequential to the administration of (ii) the anti-CTLA4 antibody, and (iii) optionally of the anti-PD-1 antibody or anti-PD-L1 antibody.

[0106] In the methods of the disclosure, (i) the antibody-drug conjugate as disclosed herein may be administered before (ii) the anti-CTLA4 antibody, and (iii) optionally of the anti- PD-1 antibody or anti-PD-L1 antibody.

[0107] In the methods of the disclosure, (ii) the anti-CTLA4 antibody and (iii) the anti- PD-1 antibody or anti-PD-L1 antibody, when present, may be administered simultaneously.

[DESCRIPTION OF THE FIGURES] [0108] Figure 1 : shows the activity of the immunoconjugate huMAb2-3-SPDB-DM4 and the anti-muPD-1 + anti-muCTLA-4 antibody co-treatment, as single agents or in combination against subcutaneous colon MC38 syngeneic tumor model in C57BI/6 mice. The curves represent the tumor volume evolution by treatment group. The curves represent medians + or - MAD at each day for each group. Continuous line: control (untreated). Dashed/dotted line: huMab2-3-SPDB-DM4 alone. Dashed line: Combination anti- muPD1/muCTLA-4 antibodies. Lozenge: huMAb2-3-SPDB-DM4 + anti-muPD1/muCTLA-4 antibodies. Panel A: huMAb2-3-SPDB-DM4 at 25 mg/kg. Panel B: huMAb2-3-SPDB-DM4 at 15 mg/kg.

[0109] Figure 2: Activity of the immunoconjugate huMAb2-3-SPDB-DM4 and the anti- mu/huPD-L1 + anti-muCTLA-4 antibody co-treatment, as single agents or in combination against subcutaneous colon MC38 syngeneic tumor model in C57BI/6 mice. Tumor volume evolution by treatment group. The curves represent medians + or - MAD at each day for each group. Continuous line: control (untreated). Dashed/dotted line: huMab2-3-SPDB-DM4 alone. Dashed line: Combination anti-mu/huPD-L1/mCTLA-4 antibodies. Lozenge: huMAb2-3-SPDB- DM4 + anti-mu/huPD-L1/mCTLA-4 antibodies. Panel A: huMAb2-3-SPDB-DM4 at 25 mg/kg. Panel B: huMAb2-3-SPDB-DM4 at 15 mg/kg.

[0110] Figure 3: Activity of the immunoconjugate huMAb2-3-SPDB-DM4 and anti- muCTLA-4 antibody as single agents or in combination against subcutaneous colon CT26 syngeneic tumor model in Balb/C mice. Tumor volume evolution by treatment group. The curves represent medians + or - MAD at each day for each group. Continuous line: control (untreated). Dashed/dotted line: huMab2-3-SPDB-DM4 alone. Dashed line: anti-mCTLA-4 antibody. Lozenge: huMAb2-3-SPDB-DM4 + anti-mCTLA-4 antibody. Panel A: huMAb2-3- SPDB-DM4 at 35 mg/kg. Panel B: huMAb2-3-SPDB-DM4 at 25 mg/kg.

[0111] Figure 4: Activity of the immunoconjugate huMAb2-3-SPDB-DM4 and the anti- mu/huPD-L1 + anti-mPD1 antibody co-treatment, as single agents or in combination against subcutaneous colon CT26 syngeneic tumor model in Balb/C mice. Tumor volume evolution by treatment group. The curves represent medians + or - MAD at each day for each group. Continuous line: control (untreated). Dashed/dotted line: huMab2-3-SPDB-DM4 alone. Dashed line: Combination anti-mPD1/mCTLA-4 antibodies. Lozenge: huMAb2-3-SPDB-DM4 + anti-mPD1/mCTLA-4 antibodies. Panel A: huMAb2-3-SPDB-DM4 at 25 mg/kg. Panel B: huMAb2-3-SPDB-DM4 at 15 mg/kg.

[0112] Figure 5: Activity of the immunoconjugate huMAb2-3-SPDB-DM4 and the anti- mu/huPD-L1 + anti-muCTLA-4 antibody co-treatment, as single agents or in combination against subcutaneous colon CT26 syngeneic tumor model in Balb/C mice. Tumor volume evolution by treatment group. The curves represent medians + or - MAD at each day for each group. Continuous line: control (untreated). Dashed/dotted line: huMab2-3-SPDB-DM4 alone. Dashed line: Combination anti-mu/huPD-L1/mCTLA-4 antibodies. Lozenge: huMAb2-3-SPDB- DM4 + anti-mu/huPD-L1/mCTLA-4 antibodies. Panel A: huMAb2-3-SPDB-DM4 at 25 mg/kg. Panel B: huMAb2-3-SPDB-DM4 at 15 mg/kg.

[BRIEF DESCRIPTION OF THE SEQUENCES]

[0001] SEQ ID NO: 1 -5 show the sequences CDR-H1 , CDR-H2, CDR-H3, CDR-L1 and CDR-L3 of the anti-CEACAM5-antibody (huMAb2-3).

[0002] SEQ ID NO: 6 shows the sequence of the variable domain of the heavy chain (VH) of the anti-CEACAM5-antibody (huMAb2-3).

[0003] SEQ ID NO: 7 shows the sequence of the variable domain of the light chain (VL) of the anti-CEACAM5-antibody (huMAb2-3).

[0004] SEQ ID NO: 8 shows the heavy chain sequence of the anti-CEACAM5-antibody (huMAb2-3).

[0005] SEQ ID NO: 9 shows the light chain sequence of the anti-CEACAM5-antibody (huMAb2-3).

[DETAILED DESCRIPTION]

Definitions

[0006] Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure shall have the meanings that are commonly understood by those of ordinary skill in the art. For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd ed., 1999, Academic Press; and the Oxford Dictionary Of Biochemistry And Molecular Biology, Revised, 2000, Oxford University Press, may provide one of skill with a general dictionary of many of the terms used in this disclosure. Exemplary methods and materials are described below, although methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure. In case of conflict, the present specification, including definitions, will control. Generally, nomenclature used in connection with, and techniques of, cell and tissue culture, molecular biology, virology, immunology, microbiology, genetics, analytical chemistry, synthetic organic chemistry, medicinal and pharmaceutical chemistry, and protein and nucleic acid chemistry and hybridization described herein are those well-known and commonly used in the art. Enzymatic reactions and purification techniques are performed according to manufacturer’s specifications, as commonly accomplished in the art or as described herein. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular.

[0007] Units, prefixes, and symbols are denoted in their International System Units (Systeme International des Unites (SI)) accepted form. Numeric ranges are inclusive of the numbers defining the range. Unless otherwise indicated, amino acid sequences are written left to right in amino to carboxy orientation. The headings provided herein are not limitations of the various aspects of the disclosure. Accordingly, the terms defined immediately below are more fully defined by reference to the specification in its entirety.

[0008] All publications and other references mentioned herein are incorporated by reference in their entirety. Although a number of documents are cited herein, this citation does not constitute an admission that any of these documents forms part of the common general knowledge in the art.

[0009] Throughout this specification and embodiments, the words “have” and “comprise,” or variations such as “has,” “having,” “comprises,” or “comprising,” will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers. It is understood that wherever aspects are described herein with the language "comprising," otherwise analogous aspects described in terms of "consisting of" and/or "consisting essentially of" are also provided.

[0010] It is to be noted that the term "a" or "an" entity refers to one or more of that entity; for example, "a nucleotide sequence," is understood to represent one or more nucleotide sequences. As such, the terms "a" (or "an"), "one or more," and "at least one" can be used interchangeably herein.

[0011] Furthermore, "and/or" where used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other. Thus, the term "and/or" as used in a phrase such as "A and/or B" herein is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Likewise, the term "and/or" as used in a phrase such as "A, B, and/or C" is intended to encompass each of the following aspects: 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).

[0012] The term “approximately” or "about" is used herein to mean approximately, roughly, around, or in the regions of. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. In general, the term "about" can modify a numerical value above and below the stated value by a variance of, e.g., 10 percent, up or down (higher or lower). In some embodiments, the term indicates deviation from the indicated numerical value by ±10%, ±5%, ±4%, ±3%, ±2%, ±1 %, ±0.9%, ±0.8%, ±0.7%, ±0.6%, ±0.5%, ±0.4%, ±0.3%, ±0.2%, ±0.1%, ±0.05%, or ±0.01 %. In some embodiments, “about” indicates deviation from the indicated numerical value by ±10%. In some embodiments, “about” indicates deviation from the indicated numerical value by ±5%. In some embodiments, “about” indicates deviation from the indicated numerical value by ±4%. In some embodiments, “about” indicates deviation from the indicated numerical value by ±3%. In some embodiments, “about” indicates deviation from the indicated numerical value by ±2%. In some embodiments, “about” indicates deviation from the indicated numerical value by ±1%. In some embodiments, “about” indicates deviation from the indicated numerical value by ±0.9%. In some embodiments, “about” indicates deviation from the indicated numerical value by ±0.8%. In some embodiments, “about” indicates deviation from the indicated numerical value by ±0.7%. In some embodiments, “about” indicates deviation from the indicated numerical value by ±0.6%. In some embodiments, “about” indicates deviation from the indicated numerical value by ±0.5%. In some embodiments, “about” indicates deviation from the indicated numerical value by ±0.4%. In some embodiments, “about” indicates deviation from the indicated numerical value by ±0.3%. In some embodiments, “about” indicates deviation from the indicated numerical value by ±0.1%. In some embodiments, “about” indicates deviation from the indicated numerical value by ±0.05%. In some embodiments, “about” indicates deviation from the indicated numerical value by ±0.01%.

[0013] An “antibody” may be a natural or conventional antibody in which two heavy chains are linked to each other by disulfide bonds and each heavy chain is linked to a light chain by a disulfide bond. There are two types of light chain, lambda (I) and kappa (k). There are five main heavy chain classes (or isotypes) which determine the functional activity of an antibody molecule: IgM, IgD, IgG, IgA and IgE. Each chain contains distinct sequence domains. The light chain includes two domains or regions, a variable domain (VL) and a constant domain (CL). The heavy chain includes four domains, a variable domain (VH) and three constant domains (CH1 , CH2 and CH3, collectively referred to as CH). The variable regions of both light (VL) and heavy (VH) chains determine binding recognition and specificity to the antigen. The constant region domains of the light (CL) and heavy (CH) chains confer important biological properties, such as antibody chain association, secretion, trans-placental mobility, complement binding, and binding to Fc receptors (FcR). The Fv fragment is the N- terminal part of the Fab fragment of an immunoglobulin and consists of the variable portions of one light chain and one heavy chain. The specificity of the antibody resides in the structural complementarity between the antibody combining site and the antigenic determinant. Antibody combining sites are made up of residues that are primarily from the hypervariable or complementarity determining regions (CDRs). Occasionally, residues from non-hypervariable or framework regions (FR) influence the overall domain structure and hence the combining site. Complementarity Determining Regions or CDRs therefore refer to amino acid sequences which together define the binding affinity and specificity of the natural Fv region of a native immunoglobulin binding site. The light and heavy chains of an immunoglobulin each have three CDRs, designated CDR1 -L, CDR2-L, CDR3-L and CDR1 -H, CDR2-H, CDR3-H, respectively. A conventional antibody antigen-binding site, therefore, includes six CDRs, comprising the CDR set from each of a heavy and a light chain V region.

[0014] As used herein, the term “antibody” intends to refer to conventional antibodies and fragments thereof, as well as single domain antibodies and fragments thereof, in particular variable heavy chain of single domain antibodies, and chimeric, humanized, bispecific or multispecific antibodies. Fragments of antibody considered herein are antigen-binding fragments.

[0015] “Framework Regions” (FRs) refer to amino acid sequences interposed between CDRs, i.e., to those portions of immunoglobulin light and heavy chain variable regions that are relatively conserved among different immunoglobulins in a single species. The light and heavy chains of an immunoglobulin each have four FRs, designated FR1 -L, FR2-L, FR3- L, FR4-L, and FR1 -H, FR2-H, FR3-H, FR4-H, respectively. A human framework region is a framework region that is substantially identical (about 85%, or more, in particular 90%, 95%, 97%, 99% or 100%) to the framework region of a naturally occurring human antibody.

[0016] In the context of the disclosure, CDR/FR definition in an immunoglobulin light or heavy chain is to be determined based on IMGT definition (Lefranc et al. Dev. Comp. Immunol., 2003, 27(1 ):55-77; www.imgt.org).

[0017] As used herein, antibody or immunoglobulin also includes “single domain antibodies” which have been more recently described and which are antibodies whose complementary determining regions are part of a single domain polypeptide. Examples of single domain antibodies include heavy chain antibodies, antibodies naturally devoid of light chains, single domain antibodies derived from conventional four-chain antibodies, engineered single domain antibodies. Single domain antibodies may be derived from any species including, but not limited to mouse, human, camel, llama, goat, rabbit, bovine. Single domain antibodies may be naturally occurring single domain antibodies known as heavy chain antibody devoid of light chains. In particular, camelidae species, for example camel, dromedary, llama, alpaca and guanaco, produce heavy chain antibodies naturally devoid of light chain. Camelid heavy chain antibodies also lack the CH1 domain.

[0018] The variable heavy chain of these single domain antibodies devoid of light chains are known in the art as “VHH” or “Nanobody®”. Similar to conventional VH domains, VHHs contain four FRs and three CDRs. VHH have advantages over conventional antibodies: they are about ten times smaller than IgG molecules, and as a consequence properly folded functional VHH can be produced by in vitro expression while achieving high yield. Furthermore, VHH are very stable, and resistant to the action of proteases. The properties and production of VHH have been reviewed by Harmsen and De Haard HJ (Appl. Microbiol. Biotechnol. 2007 Nov;77(1 ):13-22).

[0019] The term "monoclonal antibody" or “mAb” as used herein refers to an antibody molecule of a single amino acid sequence, which is directed against a specific antigen, and is not to be construed as requiring production of the antibody by any particular method. A monoclonal antibody may be produced by a single clone of B cells or hybridoma, but may also be recombinant, i.e., produced by protein engineering.

[0020] The term "humanized antibody" refers to an antibody which is wholly or partially of non-human origin, and which has been modified to replace certain amino acids, in particular in the framework regions of the VH and VL domains, in order to avoid or minimize an immune response in humans. The constant domains of a humanized antibody are most of the time human CH and CL domains.

[0021] “Fragments” of (conventional) antibodies comprise a portion of an intact antibody, in particular 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. A fragment of a conventional antibody may also be a single domain antibody, such as a heavy chain antibody or VHH.

[0022] The term “Fab” denotes an antibody fragment having a molecular weight of about 50,000 and antigen binding activity, in which about a half of the N-terminal side of the heavy chain and the entire light chain are bound together through a disulfide bond. It is usually obtained among fragments by treating IgG with a protease, such as papain.

[0023] The term “F(ab')2” refers to an antibody fragment having a molecular weight of about 100,000 and antigen binding activity, which is slightly larger than 2 identical Fab fragments bound via a disulfide bond of the hinge region. It is usually obtained among fragments by treating IgG with a protease, such as pepsin. [0024] The term “Fab'“ refers to an antibody fragment having a molecular weight of about 50,000 and antigen binding activity, which is obtained by cutting a disulfide bond of the hinge region of the F(ab')2.

[0025] A single chain Fv ("scFv") polypeptide is a covalently linked VH::VL heterodimer which is usually expressed from a gene fusion including VH and VL encoding genes linked by a peptide-encoding linker. The human scFv fragment of the disclosure includes CDRs that are held in appropriate conformation, in particular by using gene recombination techniques. Divalent and multivalent antibody fragments can form either spontaneously by association of monovalent scFvs, or can be generated by coupling monovalent scFvs by a peptide linker, such as divalent sc(Fv)2. “dsFv” is a VH::VL heterodimer stabilized by a disulphide bond. “(dsFv)2” denotes two dsFv coupled by a peptide linker.

[0026] The term “bispecific antibody” or “BsAb” denotes an antibody which combines the antigen-binding sites of two antibodies within a single molecule. Thus, BsAbs are able to bind two different antigens simultaneously. Genetic engineering has been used with increasing frequency to design, modify, and produce antibodies or antibody derivatives with a desired set of binding properties and effector functions as described for instance in EP 2 050 764 A1 .

[0027] The term “multispecific antibody” denotes an antibody which combines the antigen-binding sites of two or more antibodies within a single molecule.

[0028] The term "diabodies" refers to small antibody fragments with two antigenbinding sites, which fragments comprise a heavy-chain variable domain (VH) connected 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 of the same chain, the domains are forced to pair with the complementary domains of another chain and create two antigenbinding sites.

[0029] An amino acid sequence “at least 85% identical to a reference sequence” is a sequence having, on its entire length, 85%, or more, in particular 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity with the entire length of the reference amino acid sequence.

[0030] A percentage of “sequence identity” between amino acid sequences may be determined by comparing the two sequences, optimally aligned over a comparison window, wherein the portion of the polynucleotide or polypeptide sequence in the comparison window may comprise additions or deletions (i.e., gaps) as compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions at which the identical nucleic acid base or amino acid residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the result by 100 to yield the percentage of sequence identity. Optimal alignment of sequences for comparison is conducted by global pairwise alignment, e.g., using the algorithm of Needleman and Wunsch J. Mol. Biol. 48:443 (1970). The percentage of sequence identity can be readily determined for instance using the program Needle, with the BLOSUM62 matrix, and the following parameters gap-open=10, gap- extend=0.5.

[0031] A "conservative amino acid substitution" is one in which an amino acid residue is substituted by another amino acid residue having a side chain R group with similar chemical properties (e.g., charge, size or hydrophobicity). In general, a conservative amino acid substitution will not substantially change the functional properties of a protein. Examples of groups of amino acids that have side chains with 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 acids substitution groups can also be defined on the basis of amino acid size.

[0032] By "purified" and "isolated" it is meant, when referring to a polypeptide (i.e., the antibody of the disclosure) or a nucleotide sequence, that the indicated molecule is present in the substantial absence of other biological macromolecules of the same type. The term "purified" as used herein in particular means at least 75%, 85%, 95%, or 98% by weight, of biological macromolecules of the same type are present. An “isolated” nucleic acid molecule which encodes a particular polypeptide refers to a nucleic acid molecule which is substantially free of other nucleic acid molecules that do not encode the subject polypeptide; however, the molecule may include some additional bases or moieties which do not deleteriously affect the basic characteristics of the composition.

[0033] As used herein, the term “subject” or “patient" denotes a mammal, such as a rodent, a feline, a canine, and a primate. In particular, a subject according to the disclosure is a human.

[0034] "Administer" or "administering," as used herein refers to delivering to a subject a composition described herein. The composition can be administered to a subject using methods known in the art. In particular, the composition can be administered intravenously, subcutaneously, intramuscularly, intradermally, or via any mucosal surface, e.g., orally, sublingually, buccally, nasally, rectally, vaginally or via pulmonary route. In some embodiments, the administration is intravenous. In some embodiments, the administration is subcutaneous.

[0035] The terms “treat” or “treatment” or “therapy” refers to the administration of a compound or a composition according to the disclosure with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve, or affect a disorder, the symptoms of the condition, or to prevent or delay the onset of the symptoms, complications, or otherwise arrest or inhibit further development of the disorder in a statistically significant manner. More particularly, “treating” or “treatment” includes any approach for obtaining beneficial or desired results in a subject’s cancer condition. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more cancer symptoms or conditions, diminishment or reduction of the extent of a cancer disease or of a cancer symptom, stabilizing, i.e., not worsening, the state of a cancer disease or of a cancer symptom, prevention of a cancer disease or of a cancer symptom’s spread, delay or slowing of cancer disease or cancer symptom progression, amelioration or palliation of the cancer disease state, diminishment of the reoccurrence of cancer disease, and remission, whether partial or total and whether detectable or undetectable. In other words, "treatment" as used herein includes any cure, amelioration, or reduction of a cancer disease or symptom. A “reduction” of a symptom or a disease means decreasing of the severity or frequency of the disease or symptom, or elimination of the disease or symptom.

[0036] As used herein, the terms “effective amount” refer to an amount that provides a therapeutic benefit in the treatment, prevention, or management of pathological processes considered. The specific amount that is therapeutically effective can be readily determined by an ordinary medical practitioner and may vary depending on factors such as the type and stage of pathological processes considered, the patient’s medical history and age, and the administration of other therapeutic agents.

[0037] The unit “mg/m²” indicates the amount of compound in mg per m² of subject body surface administered per dose. The person skilled in the art is aware how to determine the required amount of compound for the subject to be treated based on his body surface, which in turn may be calculated based on height and body weight.

[0038] The unit “mg/kg” indicates the amount of compound in mg per kg of subject body administered per dose. The person skilled in the art is aware how to determine the required amount of compound for the subject to be treated based on his body weight. [0039] It is appreciated that certain features of the disclosure, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosure, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination.

[0040] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure. All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and/or materials in connection with which the publications are cited.

[0041] The list of sources, ingredients, and components as described hereinafter are listed such that combinations and mixtures thereof are also contemplated and within the scope herein.

[0042] It should be understood that every maximum numerical limitation given throughout this specification includes every lower numerical limitation, as if such lower numerical limitations were expressly written herein. Every minimum numerical limitation given throughout this specification will include every higher numerical limitation, as if such higher numerical limitations were expressly written herein. Every numerical range given throughout this specification will include every narrower numerical range that falls within such broader numerical range, as if such narrower numerical ranges were all expressly written herein.

[0043] All lists of items, such as, for example, lists of ingredients, are intended to and should be interpreted as Markush groups. Thus, all lists can be read and interpreted as items “selected from the group consisting of’ the list of items “and combinations and mixtures thereof.”

[0044] Referenced herein may be trade names for components including various ingredients utilized in the present disclosure. The inventors herein do not intend to be limited by materials under any particular trade name. Equivalent materials (e.g., those obtained from a different source under a different name or reference number) to those referenced by trade name may be substituted and utilized in the descriptions herein.

Antibody-drug conjugate comprising an anti-CEACAM5-antibody [0045] The present disclosure relates to an antibody-drug conjugate (ADC) comprising an anti-CEACAM5-antibody, or a fragment thereof, which is used in combination with an anti- CTLA4 antibody, or a fragment thereof, and optionally an anti-PD-1 antibody or anti-PD-L1 antibody, or a fragment thereof, for the treatment of cancer.

[0046] The antibody-drug conjugate typically comprises an anti-CEACAM5-antibody and at least one chemotherapeutic agent. An antibody-drug conjugate (ADC) comprises an anti-CEACAM5-antibody conjugated to at least one chemotherapeutic agent. In the antibodydrug conjugate, the anti-CEACAM5-antibody may be covalently attached via a cleavable or non-cleavable linker to at least one chemotherapeutic agent.

Antl-CEA CAM5-antibody

[0047] According to an embodiment, the antibody-drug conjugate comprises an anti- CEACAM5-antibody or a fragment thereof.

[0048] According to an embodiment, the antibody-drug conjugate comprises a humanized anti-CEACAM5-antibody or a fragment thereof.

[0049] According to an embodiment, the anti-CEACAM5-antibody or a fragment thereof comprises a CDR-H1 consisting of SEQ ID NO: 1 , CDR-H2 consisting of SEQ ID NO: 2, CDR-H3 consisting of SEQ ID NO: 3, CDR-L1 consisting of SEQ ID NO: 4, CDR-L2 consisting of amino acid sequence NTR, and CDR-L3 consisting of SEQ ID NO: 5.

[0050] In a further embodiment, the anti-CEACAM5-antibody or a fragment thereof comprises a variable domain of a heavy chain (VH) having at least 90% identity to SEQ ID NO: 6, and a variable domain of a light chain (VL) having at least 90% identity to SEQ ID NO: 7.

[0051] In a further embodiment, the anti-CEACAM5-antibody or a fragment thereof comprises a variable domain of a heavy chain (VH) having at least 90% identity to SEQ ID NO: 6, and a variable domain of a light chain (VL) having at least 90% identity to SEQ ID NO: 7, wherein CDR1 -H consists of SEQ ID NO: 1 , CDR2-H consists of SEQ ID NO: 2, CDR3-H consists of SEQ ID NO: 3, CDR1 -L consists of SEQ ID NO: 4, CDR2-L consists of amino acid sequence NTR, and CDR3-L consists of SEQ ID NO: 5.

[0052] In a further embodiment, the anti-CEACAM5-antibody or a fragment thereof comprises a variable domain of a heavy chain (VH) having at least 92%, at least 95%, at least 98% identity to SEQ ID NO: 6, and a variable domain of a light chain (VL) having at least 92%, at least 95%, at least 98% identity to SEQ ID NO: 7, wherein CDR1 -H consists of SEQ ID NO: 1 , CDR2-H consists of SEQ ID NO: 2, CDR3-H consists of SEQ ID NO: 3, CDR1 -L consists of SEQ ID NO: 4, CDR2-L consists of amino acid sequence NTR, and CDR3-L consists of SEQ ID NO: 5.

[0053] In a further embodiment, the anti-CEACAM5-antibody or a fragment thereof comprises a variable domain of a heavy chain (VH) consisting of SEQ ID NO: 6 and a variable domain of a light chain (VL) consisting of SEQ ID NO: 7.

[0054] The anti-CEACAM5-antibody or a fragment thereof comprises in a further embodiment:

[0055] - a variable domain of heavy chain consisting of sequence EVQLQESGPGLVKPGGSLSLSCAASGFVFSSYDMSWVRQTPERGLEWVAYISSGGGITYA PSTVKGRFTVSRDNAKNTLYLQMNSLTSEDTAVYYCAAHYFGSSGPFAYWGQGTLVTVSS (SEQ ID NO: 6, with CDRs shown in bold characters) in which FR1 -H spans amino acid positions 1 to 25, CDR1 -H spans amino acid positions 26 to 33 (SEQ ID NO: 1 ), FR2-H spans amino acid positions 34 to 50, CDR2-H spans amino acid positions 51 to 58 (SEQ ID NO: 2), FR3-H spans amino acid positions 59 to 96, CDR3-H spans amino acid positions 97 to 109 (SEQ ID NO: 3), and FR4-H spans amino acid positions 1 10 to 120, and

[0056] - a variable domain of light chain consisting of sequence DIQMTQSPASLSASVGDRVTITCRASENIFSYLAWYQQKPGKSPKLLVYNTRTLAEGVPSFS GSGSGTDFSLTISSLQPEDFATYYCQHHYGTPFTFGSGTKLEIK (SEQ ID NO: 7, with CDRs shown in bold characters) in which FR1 -L spans amino acid positions 1 to 26, CDR1 -L spans amino acid positions 27 to 32 (SEQ ID NO: 4), FR2-L spans amino acid positions 33 to 49, CDR2-L spans amino acid positions 50 to 52, FR3-L spans amino acid positions 53 to 88, CDR3-L spans amino acid positions 89 to 97 (SEQ ID NO: 5), and FR4-L spans amino acid positions 98 to 107.

[0057] In a further embodiment, the anti-CEACAM5-antibody or a fragment thereof comprises a heavy chain (HC) having at least 90% sequence identity to SEQ ID NO: 8 and a light chain (LC) having at least 90% sequence identity to SEQ ID NO: 9.

[0058] In a further embodiment, the anti-CEACAM5-antibody or a fragment thereof comprises a heavy chain (HC) having at least 90% sequence identity to SEQ ID NO: 8, and a light chain (LC) having at least 90% sequence identity to SEQ ID NO: 9, wherein CDR1 -H consists of SEQ ID NO: 1 , CDR2-H consists of SEQ ID NO: 2, CDR3-H consists of SEQ ID NO: 3, CDR1 -L consists of SEQ ID NO: 4, CDR2-L consists of amino acid sequence NTR, and CDR3-L consists of SEQ ID NO: 5.

[0059] In a further embodiment, the anti-CEACAM5-antibody or a fragment thereof comprises a heavy chain (HC) having at least 92%, at least 95%, at least 98% identity to SEQ ID NO: 8 and a light chain (LC) having at least 92%, at least 95%, at least 98% identity to SEQ ID NO: 9, wherein CDR1 -H consists of SEQ ID NO: 1 , CDR2-H consists of SEQ ID NO: 2, CDR3-H consists of SEQ ID NO: 3, CDR1 -L consists of SEQ ID NO: 4, CDR2-L consists of amino acid sequence NTR, and CDR3-L consists of SEQ ID NO: 5.

[0060] In a further embodiment, the anti-CEACAM5-antibody or a fragment thereof comprises a heavy chain (HC) consisting of SEQ ID NO: 8 and a light chain (LC) consisting of SEQ ID NO: 9.

[0061] The anti-CEACAM5-antibody may also be a single domain antibody or a fragment thereof. In particular, a single domain antibody fragment may consist of a variable heavy chain (VHH) which comprises the CDR1 -H, CDR2-H and CDR3-H of the antibodies as described above. The antibody may also be a heavy chain antibody, i.e., an antibody devoid of light chain, which may or may not contain a CH1 domain.

[0062] The single domain antibody or a fragment thereof may also comprise the framework regions of a camelid single domain antibody, and optionally the constant domain of a camelid single domain antibody.

[0063] The anti-CEACAM5-antibody may also be an antibody fragment, in particular a humanized antibody fragment, selected from the group consisting of Fv, Fab, F(ab')2, Fab', dsFv, (dsFv)2, scFv, sc(Fv)2, and diabodies.

[0064] The antibody may also be a bispecific or multispecific antibody formed from antibody fragments, at least one antibody fragment being an antibody fragment according to the disclosure. Multispecific antibodies are polyvalent protein complexes as described for instance in EP 2 050 764 A1 or US 2005/0003403 A1 .

[0065] The anti-CEACAM5-antibody and fragments thereof can be produced by any technique well known in the art. In particular, said antibodies are produced by techniques as hereinafter described.

[0066] The anti-CEACAM5-antibody and fragments thereof can be used in an isolated (e.g., purified) from or contained in a vector, such as a membrane or lipid vesicle (e.g., a liposome).

[0067] The anti-CEACAM5-antibody and fragments thereof may be produced by any technique known in the art, such as, without limitation, any chemical, biological, genetic, or enzymatic technique, either alone or in combination.

[0068] Knowing the amino acid sequence of the desired sequence, one skilled in the art can readily produce the anti-CEACAM5-antibody and fragments thereof, by standard techniques for production of polypeptides. For instance, they can be synthesized using well- known solid phase method, in particular using a commercially available peptide synthesis apparatus (such as that made by Applied Biosystems, Foster City, California) and following the manufacturer’s instructions. Alternatively, anti-CEACAM5-antibody and fragments thereof 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 incorporation of DNA sequences encoding the desired (poly)peptide into expression vectors and introduction of such vectors into suitable eukaryotic or prokaryotic hosts that will express the desired polypeptide, from which they can be later isolated using well-known techniques.

[0069] Anti-CEACAM5-antibody and fragments thereof are suitably separated from the culture medium by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxyapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.

[0070] Methods for producing humanized antibodies based on conventional recombinant DNA and gene transfection techniques are well known in the art (See, e. g., Riechmann L. et al. 1988; Neuberger MS. et al. 1985). Antibodies can be humanized using a variety of techniques known in the art including, for example, the technique disclosed in the application W02009/032661 , CDR-grafting (EP 239,400; PCT publication WO91/09967; U.S. Pat. Nos. 5,225,539; 5,530,101 ; and 5,585,089), veneering or resurfacing (EP 592,106; EP 519,596; Padlan EA (1991 ); Studnicka GM et al. (1994); Roguska MA. et al. (1994)), and chain shuffling (U.S. Pat. No.5, 565, 332). The general recombinant DNA technology for preparation of such antibodies is also known (see European Patent Application EP 125023 and International Patent Application WO 96/02576).

[0071 ] The Fab of the anti-CEACAM5-antibody can be obtained by treating an antibody which specifically reacts with CEACAM5 with a protease, such as papain. Also, the Fab of the anti-CEACAM5-antibody can be produced by inserting DNA sequences encoding both chains of the Fab of the anti-CEACAM5-antibody into a vector for prokaryotic expression, or for eukaryotic expression, and introducing the vector into prokaryotic or eukaryotic cells (as appropriate) to express the Fab of the anti-CEACAM5-antibody.

[0072] The F(ab')2 of the anti-CEACAM5-antibody can be obtained treating an antibody which specifically reacts with CEACAM5 with a protease, pepsin. Also, the F(ab')2 of the anti-CEACAM5-antibody can be produced by binding Fab' described below via a thioether bond or a disulfide bond. [0073] The Fab' of the anti-CEACAM5-antibody can be obtained treating F(ab')2 which specifically reacts with CEACAM5 with a reducing agent, such as dithiothreitol . Also, the Fab' of the anti-CEACAM5-antibody can be produced by inserting DNA sequences encoding Fab' chains of the antibody into a vector for prokaryotic expression, or a vector for eukaryotic expression, and introducing the vector into prokaryotic or eukaryotic cells (as appropriate) to perform its expression.

[0074] The scFv of the anti-CEACAM5-antibody can be produced by taking sequences of the CDRs or VH and VL domains as previously described, constructing a DNA encoding an scFv fragment, inserting the DNA into a prokaryotic or eukaryotic expression vector, and then introducing the expression vector into prokaryotic or eukaryotic cells (as appropriate) to express the scFv. To generate a humanized scFv fragment, a well-known technology called CDR grafting may be used, which involves selecting the complementary determining regions (CDRs) according to the disclosure and grafting them onto a human scFv fragment framework of known three-dimensional structure (see, e. g., W098/45322; WO 87/02671 ; US5,859,205; US5,585,089; US4, 816,567; EP0173494).

[0075] In an embodiment, the anti-CEACAM5 antibody is tusamitamab (CAS [2349294-95-5],

Chemotherapeutic agents

[0076] The antibody-drug conjugate for the use according to the present disclosure typically comprises at least one chemotherapeutic agent (also referred herein to cytotoxic agent). A chemotherapeutic agent as used herein refers to an agent that kills cells, including cancer cells. Such agents favorably stop cancer cells from dividing and growing and cause tumors to shrink in size. The expression “chemotherapeutic agent” is used herein interchangeably with the expressions “cytotoxic agent”, “growth inhibitory agent” or “cytostatic drug”.

[0077] The term "chemotherapeutic agent" as used herein refers to a substance that inhibits or prevents the function of cells and/or causes destruction of cells. The term “chemotherapeutic agent" is intended to include radioisotopes, enzymes, antibiotics, and toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof, and the various antitumor or anticancer agents disclosed below. In some embodiments, the chemotherapeutic agent is an antimetabolite. [0078] In a further embodiment, the chemotherapeutic agent is selected from the group consisting of radioisotopes, protein toxins, small molecule toxins, and combinations thereof.

[0079] Radioisotopes include radioactive isotopes suitable for treating cancer. Such radioisotopes generally emit mainly beta-radiation. In a further embodiment, the radioisotopes are selected from the group consisting of At²¹¹, Bi²¹², Er¹⁶⁹, 1¹³¹ , 1¹²⁵, Y⁹⁰, In¹¹¹ , P³², Re¹⁸⁶, Re¹⁸⁸, Sm¹⁵³, Sr⁸⁹, radioactive isotopes of Lu, and combinations thereof. In an embodiment, the radioactive isotope is alpha-emitter isotope, more specifically Th²²⁷, which emits alpharadiation.

[0080] In a further embodiment, the small molecule toxins are selected from antimetabolites, DNA-alkylating agents, DNA-cross-linking agents, DNA-intercalating agents, antimicrotubule agents, topoisomerase inhibitors, and combinations thereof.

[0081] In a further embodiment, the anti-microtubule agent is selected from the group consisting of taxanes, vinca alkaloids, maytansinoids, colchicine, podophyllotoxin, gruseofulvin, and combinations thereof.

[0082] In some embodiment a chemotherapeutic agent may be a maytansinoid.

[0083] According to an embodiment, maytansinoids are selected from maytansinol, maytansinol analogs, and combinations thereof.

[0084] Examples of suitable maytansinol analogues include those having a modified aromatic ring and those having modifications at other positions. Such suitable maytansinoids are disclosed 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; 4,362,663; 4,364,866; 4,450,254; 4,322,348; 4,371 ,533; 6,333,410; 5,475,092; 5,585,499; and 5,846,545.

[0085] In a further embodiment, the cytotoxic conjugates of the present disclosure utilize the thiol-containing maytansinoid (DM1 ), formally termed A/2’-deacetyl-/\/2’-(3-mercapto- 1 -oxopropyl)-maytansine, as the cytotoxic agent. DM1 is represented by the following structural formula (I):

[0086] In a further embodiment, the cytotoxic conjugates of the present disclosure utilize the thiol-containing maytansinoid DM4, formally termed A/2’-deacetyl-/V-2’(4-methyl-4- mercapto-1 -oxopentyl)-maytansine, as the cytotoxic agent. DM4 is represented by the following structural formula (II):

[0087] In further embodiments of the disclosure other maytansines, including thiol and disulfide-containing maytansinoids bearing a mono or di-alkyl substitution on the carbon atom bearing the sulfur atom, may be used. These include a maytansinoid having, at C-3, C-14 hydroxymethyl, C-15 hydroxy, or C-20 desmethyl, an acylated amino acid side chain with an acyl group bearing a hindered sulfhydryl group, wherein the carbon atom of the acyl group bearing the thiol functionality has one or two substituents, said substituents being CH₃, C2H5, linear or branched alkyl or alkenyl having from 1 to 10 reagents and any aggregate which may be present in the solution.

[0088] Examples of these cytotoxic agents and of methods of conjugation are further given in the application WO 2008/010101 which is incorporated by reference. [0089] The immunoconjugates according to the present disclosure can be prepared as described in the application WO 2004/091668, the entire content of which is incorporated herein by reference.

[0090] Accordingly, in a further embodiment, the maytansinoids are selected from the group consisting of A/2’-deacetyl-/\/2’-(3-mercapto-1 -oxopropyl)-maytansine (DM1) or N2’- deacetyl-/V-2’(4-methyl-4-mercapto-1 -oxopentyl)-maytansine (DM4), and combinations thereof.

[0091] In a further embodiment, in the antibody-drug conjugate, the anti-CEACAM5- antibody is covalently attached via a cleavable or non-cleavable linker to the at least one chemotherapeutic agent.

[0092] In a further embodiment, the linker is selected from the group consisting of N- succinimidyl pyridyldithiobutyrate (SPDB), 4-(pyridin-2-yldisulfanyl)-2-sulfo-butyric acid (sulfo- SPDB), and succinimidyl(N-maleimidomethyl) cyclohexane-1 -carboxylate (SMCC).

[0093] In a further embodiment, the linker binds to a lysine or cysteine residue in the Fc region of the anti-CEACAM5 antibody. In a further embodiment, the linker forms a disulfide bond or a thioether bond with the maytansine.

[0094] In particular, the anti-CEACAM5-antibody-drug conjugate may be selected from the group consisting of:

[0095] the anti-CEACAM5-SPDB-DM4-antibody-drug conjugate of formula (III):

[0097] and

[0098] an anti-CEACAM5-SMCC-DM1 -antibody-drug conjugate of formula (V):

[0099] In formulas (III), (IV) and (V) above, “n” corresponds to the number of molecules of chemotherapeutic agent conjugated per molecule of antibody. It corresponds to the “drug- to-antibody ratio” (or “DAR”) defined below and may range from 1 to 10.

[0100] In a further embodiment, the antibody-drug conjugate of the present disclosure comprises an anti-CEACAM5-antibody, which comprises a heavy chain (HC) of SEQ ID NO: 8 and a light chain (CL) of SEQ ID NO: 9 (tusamitamab), wherein tusamitamab is covalently linked to N2’-deacetyl-N-2’(4-methyl-4-mercapto-1 -oxopentyl)-maytansine (DM4) via N- succinimidyl pyridyldithiobutyrate (SPDB). Thereby, the antibody-drug conjugate tusamitamab ravtansine (huMAb2-3-SPDB-DM4) is obtained.

[0101] In an embodiment, the antibody-drug conjugate of the present disclosure is tusamitamab ravtansine (CAS [2254086-60-5]).

[0102] “Linker”, as used herein, means a chemical moiety comprising a covalent bond or a chain of atoms that covalently attaches the antibody to the chemotherapeutic agent moiety (e.g., a cytostatic agent, a cytotoxic agent or a growth inhibitory agent). Suitable linkers are well known in the art and include disulfide groups, thioether groups, acid labile groups, photolabile groups, peptidase labile groups and esterase labile groups.

[0103] The conjugates may be prepared by in vitro methods. In order to link a drug or prodrug to the antibody, e.g., a chemotherapeutic agent, a linking group is used. Suitable linking groups are well known in the art and include disulfide groups, thioether groups, acid labile groups, photolabile groups, peptidase labile groups and esterase labile groups. Conjugation of an antibody with a chemotherapeutic agent of the disclosure, such as a cytotoxic agent, may be made using a variety of bifunctional protein coupling agents including but not limited to N-succinimidyl pyridyldithiobutyrate (SPDB), butanoic acid 4-[(5-nitro-2- pyridinyl)dithio]-2,5-dioxo- 1 -pyrrolidinyl ester (nitro-SPDB), 4-(pyridin-2-yldisulfanyl)-2-sulfo- butyric acid (sulfo-SPDB), N-succinimidyl (2-pyridyldithio) propionate (SPDP), succinimidyl (N- maleimidomethyl) cyclohexane-1 -carboxylate (SMCC), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCL), active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bis-azido compounds (such as bis (p-azidobenzoyl)-hexanediamine), bis-diazonium derivatives (such as bis-(p- diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as toluene 2,6-diisocyanate), and bis-active fluorine compounds (such as 1 ,5-difluoro-2,4- dinitrobenzene). For example, a ricin immunotoxin can be prepared as described in Vitetta et al (1987). Carbon labeled 1 - isothiocyanatobenzyl methyldiethylene triaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for conjugation of radionucleotide to the antibody (WO 94/1 1026).

[0104] The linker may be a "cleavable linker" facilitating release of the chemotherapeutic agent in the cell. For example, an acid-labile linker, a peptidase-sensitive linker, an esterase labile linker, a photolabile linker or a disulfide-containing linker (See e.g., U.S. Patent No. 5,208,020) may be used. The linker may be also a "non-cleavable linker" (for example SMCC linker) that might led to better tolerance in some cases.

[0105] In general, the conjugate can be obtained by a process comprising the steps of:

[0106] (i) bringing into contact an optionally-buffered aqueous solution of a cell-binding agent (e.g., an antibody according to the disclosure) with solutions of a linker and a chemotherapeutic agent, such as a cytotoxic compound (or agent);

[0107] (ii) then optionally separating the conjugate which was formed in (i) from the unreacted cell-binding agent (e.g., antibody of the disclosure) and unreacted chemotherapeutic agent, such as unreacted cytotoxic compound (or agent).

[0108] The aqueous solution of cell-binding agent can be buffered with buffers such as, e.g., potassium phosphate, acetate, citrate or N-2-Hydroxyethylpiperazine-N’-2- ethanesulfonic acid (Hepes buffer). The buffer depends upon the nature of the cell-binding agent (e.g., antibody of the disclosure). The chemotherapeutic agent, such as the cytotoxic compound (or agent), is in solution in an organic polar solvent, e.g., dimethyl sulfoxide (DMSO) or dimethylacetamide (DMA).

[0109] The reaction temperature is usually comprised between 20°C and 40°C. The reaction time can vary from 1 hour to 24 hours. The reaction between the cell-binding agent and the chemotherapeutic agent, such as the cytotoxic agent, can be monitored by size exclusion chromatography (SEC) with a refractometric and/or UV detector. If the conjugate yield is too low, the reaction time can be extended.

[0110] A number of different chromatography methods can be used by the person skilled in the art in order to perform the separation of step (ii): the conjugate can be purified, for example from aggregates, e.g., by SEC, adsorption chromatography (such as ion exchange chromatography, IEC), hydrophobic interaction chromatography (HIC), affinity chromatography, mixed-support chromatography such as hydroxyapatite chromatography, or high-performance liquid chromatography (HPLC). Purification by dialysis or diafiltration can also be used.

[0111] As used herein, the term “aggregates” means the associations which can be formed between two or more cell-binding agents, said agents being modified or not by conjugation. The aggregates can be formed under the influence of a great number of parameters, such as a high concentration of cell-binding agent (e.g., antibody of the disclosure) in the solution, the pH of the solution, high shearing forces, the number of bonded dimers and their hydrophobic character, the temperature (see Wang & Gosh, 2008, J. Membrane Sci., 318: 311 -316, and references cited therein); note that the relative influence of some of these parameters is not clearly established. In the case of proteins and antibodies, the person skilled in the art will refer to Cromwell et al. (2006, AAPS Journal, 8(3): E572-E579). The content in aggregates can be determined with techniques well known to the skilled person, such as SEC (see Walter et al., 1993, Anal. Biochem., 212(2): 469-480).

[0112] After step (i) or (ii), the conjugate-containing solution can be submitted to an additional step (iii) of chromatography, ultrafiltration and/or diafiltration.

[0113] The conjugate is recovered at the end of these steps in an aqueous solution.

[0114] In a further embodiment, the antibody-drug conjugate according to the disclosure is characterized by a “drug-to-antibody ratio” (or “DAR”) ranging from 1 to 10, or from 2 to 5, or from 3 to 4. This is generally the case of conjugates including maytansinoid molecules.

[0115] This DAR number can vary with the nature of the antibody and of the drug (i.e. the chemotherapeutic agent, such as a cytotoxic agent or a growth-inhibitory agent) used along with the experimental conditions used for the conjugation (like the ratio chemotherapeutic agent (e.g., growth-inhibitory agent)/antibody, the reaction time, the nature of the solvent and of the cosolvent if any).Thus the contact between the antibody and the chemotherapeutic agent, such as a cytotoxic agent or a growth-inhibitory agent, leads to a mixture comprising several conjugates differing from one another by different drug-to-antibody ratios; optionally the naked antibody; optionally aggregates. The DAR that is determined is thus a mean value.

[0116] A method which can be used to determine the DAR consists in measuring spectrophotometrically the ratio of the absorbance at of a solution of substantially purified conjugate at AD and 280 nm. 280 nm is a wavelength generally used for measuring protein concentration, such as antibody concentration. The wavelength AD is selected so as to allow discriminating the drug from the antibody, i.e., as readily known to the skilled person, AD is a wavelength at which the drug (i.e., chemotherapeutic agent) has a high absorbance and AD is sufficiently remote from 280 nm to avoid substantial overlap in the absorbance peaks of the drug and antibody. AD may be selected as being 252 nm in the case of maytansinoid molecules. A method of DAR calculation may be derived from Antony S. Dimitrov (ed), LLC, 2009, Therapeutic Antibodies and Protocols, vol 525, 445, Springer Science:

[0117] The absorbances for the conjugate at AD (AAD) and at 280 nm (A280) are measured either on the monomeric peak of the size exclusion chromatography (SEC) analysis (allowing to calculate the “DAR(SEC)” parameter) or using a classic spectrophotometer apparatus (allowing to calculate the “DAR(UV)” parameter). The absorbances can be expressed as follows:

[0118] AAD = (cD x eDAD) + (cA x eAAD)

[0119] A280 = (cD x eD280) + (cA x eA280)

[0120] wherein:

[0121] cD and cA are respectively the concentrations in the solution of the drug (i.e., chemotherapeutic agent) and of the antibody,

[0122] eDAD and eD280 are respectively the molar extinction coefficients of the drug at AD and 280 nm,

[0123] eAAD and eA280 are respectively the molar extinction coefficients of the antibody at AD and 280 nm.

[0124] Resolution of these two equations with two unknowns leads to the following equations:

[0125] cD = [(eA280 x AAD) - (eAAD x A280)] / [(eDAD x eA280) - (eAAD x eD280)]

[0126] cA = [A280 - (cD x eD280)] / eA280

[0127] The average DAR is then calculated from the ratio of the drug concentration to that of the antibody: DAR = cD / cA. ADC dosages

[0128] In some embodiments, it is disclosed an antibody-drug conjugate comprising an anti-CEACAM5-antibody and a chemotherapeutic agent for use for treating a cancer in combination with an anti-CTLA4 antibody and, when presents, an anti-PD-1 antibody or anti- PD-L1 antibody, wherein the antibody-drug conjugate may be administered at a dose of about 60 mg/m² to about 210 mg/m², or from about 80 to about 170 mg/m², or from about 100 to about 170 mg/m², or from about 120 to about 170 mg/m², or from about 135 to about 170 mg/m², or from about 150 to about 170 mg/m² of body surface area of a subject in need thereof.

[0129] In some embodiments, the antibody-drug conjugate may be administered at a dose of from about 60 to about 210 mg/m², or from about 80 to about 170 mg/m², or from about 100 to about 150 mg/m².

[0130] In some embodiments, the antibody-drug conjugate may be administered at a dose of 80 mg/m² to 210 mg/m², 80 mg/m² to 170 mg/m², or at a dose of 80 mg/m² to 150 mg/m², or at a dose of 80 mg/m² to 120 mg/m², or at a dose of 80 mg/m² to 100 mg/m².

[0131] In various embodiments, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 60, 70, 80, 90, 100, 110, 120, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, or about 210 mg/m².

[0132] In some embodiments, the antibody-drug conjugate may be administered at a dose level of 80, 100, 120, 150, 170, 180, or 210 mg/m².

[0133] In various embodiments, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m².

[0134] In some embodiments, the ADC may be administered at a dose of about 80 mg/m² to about 170 mg/m², or at a dose of about 80 mg/m² to about 150 mg/m², or at a dose of about 100 mg/m² to about 120 mg/m².

[0135] In some embodiments, the ADC may be administered at a dose of about 80 mg/m² or about 100 mg/m² or about 120 mg/m² or about 150 mg/m² or about 170 mg/m².

[0136] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 80 mg/m².

[0137] In some embodiments, the ADC may be administered at a dose of about 100 mg/m². [0138] In some embodiments, the ADC may be administered at a dose of about 120 mg/m².

[0139] In some embodiments, the ADC may be administered at a dose of about 150 mg/m².

[0140] In some embodiments, the ADC may be administered at a dose of about 170 mg/m².

[0141] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 80, 100, 120, 150 or about 170 mg/m², as a loading dose (or first dose).

[0142] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 80 mg/m², as a loading dose.

[0143] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 100 mg/m², as a loading dose.

[0144] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 120 mg/m², as a loading dose.

[0145] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 150 mg/m², as a loading dose.

[0146] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 170 mg/m², as a loading dose.

[0147] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 80, 100, 120, 150 or about 170 mg/m², as a subsequent dose (or second dose).

[0148] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 80 mg/m², as a subsequent dose.

[0149] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 100 mg/m², as a subsequent dose.

[0150] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 120 mg/m², as a subsequent dose. [0151] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 150 mg/m², as a subsequent dose.

[0152] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 170 mg/m², as a subsequent dose.

[0153] A subsequent dose may be administered on day 1 of cycle(s) subsequent to the first cycle (the subsequent or additional cycles).

[0154] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 80, 100, 120, 150 or about 170 mg/m², as a loading dose, on a first cycle of treatment, e.g., at day 1 , and then at a dose of about 80, 100, 120, 150 or about 170 mg/m², as a subsequent dose, e.g., at day 1 , on additional cycle(s).

[0155] In certain embodiments, for subjects with a body surface area (BSA) >2.2 m², the dose of the antibody-drug conjugate comprising an anti-CEACAM5-antibody may be calculated based on a BSA of 2.2 m².

[0156] In some embodiments, the dosage regimen may comprise administration of a dose over a period of about 10 minutes to about 48 hours, or of about 1 h to about 48h, such as over a period of 1 h to 4h. In some embodiments, the dosage regimen may comprise administration of a dose over a period of about 1 h.

[0157] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered over about 30 min to about 3 hours, over about 1 hour to about 2hours, or over about 1 .5 hours.

[0158] The antibody-drug conjugate may be tusamitamab ravtansine (huMAb2-3- SPDB-DM4).

Anti-CTLA4 antibody

[0159] The combination of the disclosure comprises an anti-CTLA4 antibody, or a fragment thereof, for the treatment of cancer.

[0160] The combination of the disclosure may comprise an anti-CTLA4 antibody, or a fragment thereof, for the treatment of cancer. [0161] In one embodiment, the anti-CTLA4 antibody, or a fragment thereof, is a monoclonal antibody. In one embodiment, the anti-CTLA4 antibody is an IgG.

[0162] CTLA-4 is a CD28 homologue that binds to CD80/CD86 (B7 ligands) to inhibit T-cell function. It is expressed on the surface of activated CD4-positive and CD8+ T cells, as well as on regulatory T lymphocytes (Tregs), which helps support their suppressive function. CTLA-4 mediates immuno-suppression by blocking the T-lymphocyte response, reducing the proliferation of T lymphocytes and increasing the activity of Tregs. Blocking CTLA-4 enhances an antitumor immunity by promoting T-cell activation and cytotoxic T-lymphocyte proliferation.

[0163] CTLA-4-blocking antibodies are antibodies directed against CTLA-4. By targeting CTLA-4 these antibodies prevent the interaction between the costimulatory molecules B7.1 an B7.2 (CD80 and CD86) and CTLA-4, thus removing the CTLA4 inhibitory signal and releasing a brake on the immune system.

[0164] A number of anti-CTLA4 antibodies have been approved for clinical use or are undergoing clinical trials in the treatment of cancer. These include ipilimumab (YERVOY®), tremelimumab (IMJUDO®), quavonlimab, or zalifrelimab.

[0165] In one embodiment, the anti-CTLA4 antibody is ipilimumab, tremelimumab, or durvalumab.

[0166] In some embodiments, the CTLA4 antibody may be tremelimumab or durvalumab.

[0167] In some embodiments, the CTLA4 antibody may be tremelimumab.

[0168] In some embodiments, the CTLA4 antibody may be durvalumab.

[0169] In one embodiment, the anti-CTLA4 antibody is a fully human monoclonal IgG 1 antibody against human CTLA4.

[0170] The anti-CTLA4 antibody, or a fragment thereof, may also be a single domain antibody or a fragment thereof. In particular, a single domain antibody fragment may consist of a variable heavy chain (VHH) which comprises the CDR1 -H, CDR2-H and CDR3-H of the antibodies as described above. The antibody may also be a heavy chain antibody, i.e., an antibody devoid of light chain, which may or may not contain a CH1 domain.

[0171] The single domain antibody or a fragment thereof may also comprise the framework regions of a camelid single domain antibody, and optionally the constant domain of a camelid single domain antibody. [0172] The anti-CTLA4 antibody may also be an antibody fragment, in particular a humanized antibody fragment, selected from the group consisting of Fv, Fab, F(ab')2, Fab', dsFv, (dsFv)2, scFv, sc(Fv)2, and diabodies.

[0173] The antibody may also be a bispecific or multispecific antibody formed from antibody fragments, at least one antibody fragment being an antibody fragment according to the disclosure. The anti-CTLA4 antibody and fragments thereof can be produced by any technique well known in the art. In particular, said antibodies are produced by techniques as already described.

[0174] The anti-CTLA4 antibody and fragments thereof can be used in an isolated (e.g., purified) from or contained in a vector, such as a membrane or lipid vesicle (e.g., a liposome).

[0175] The anti-CTLA4 antibody and fragments thereof may be produced by any technique known in the art, such as, without limitation, any chemical, biological, genetic, or enzymatic technique, either alone or in combination.

[0176] In some embodiments, the dosage regimen may comprise administration of a dose over a period of about 10 minutes to about 48 hours, or of about 1 h to about 48h, such as over a period of 1 h to 4h. In some embodiments, the dosage regimen may comprise administration of a dose over a period of about 1 h.

[0177] In some embodiments, the anti-CTLA4 may be administered over about 30 minutes to over about 3 hours, or over about 1 hour to about 2 hours, or over about one hour.

Antl-CTLA4 antibody dosage

[0178] The anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg to about 10 mg/kg, or at a dose of about 3 mg/kg to about 10 mg/kg of weight of a subject in need thereof.

[0179] In some embodiments, the CTLA4 antibody may be administered at a dose of about 1 mg/kg to about 10 mg/kg, or at a dose of about 3 mg/kg to about 10 mg/kg.

[0180] In some embodiments, the CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of about 6 mg/kg, or at a dose of about 8 mg/kg, or at a dose of about 10 mg/kg.

[0181 ] In some embodiments, the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg. [0182] In some embodiments, the anti-CTLA4 antibody may be administered at a dose of about 2.5 mg/kg.

[0183] In some embodiments, the anti-CTLA4 antibody may be administered at a dose of about 3 mg/kg.

[0184] In some embodiments, the anti-CTLA4 antibody may be administered at a dose of about 4 mg/kg.

[0185] In some embodiments, the anti-CTLA4 antibody may be administered at a dose of about 6 mg/kg.

[0186] In some embodiments, the anti-CTLA4 antibody may be administered at a dose of about 8 mg/kg.

[0187] In some embodiments, the anti-CTLA4 antibody may be administered at a dose of about 10 mg/kg.

[0188] According to an embodiment, the anti-CTLA4 antibody may be administered at a dose of about 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of 10 mg/kg, as a loading dose (or first dose).

[0189] According to an embodiment, the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of about 6 mg/kg, or at a dose of about 8 mg/kg, or at a dose of about 10 mg/kg, as a loading dose.

[0190] A loading dose may be administered on day 1 of the first cycle.

[0191] According to an embodiment, the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of about 6 mg/kg, or at a dose of about 8 mg/kg, or at a dose of about 10 mg/kg, as a subsequent dose (or second dose).

[0192] A subsequent dose may be administered on day 1 of cycle(s) subsequent to the first cycle (the subsequent or additional cycles).

[0193] According to an embodiment, the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of about 10 mg/kg, as a loading dose, on a first cycle of treatment, e.g., at day 1 , and then at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of about 6 mg/kg, or at a dose of about 8 mg/kg, or at a dose of about 10 mg/kg, as a subsequent dose, e.g., at day 1 , on additional cycle(s). [0194] In some embodiments, the dosage regimen may comprise administration of a dose over a period of about 10 minutes to about 48 hours, or of about 1 h to about 48h, such as over a period of 1 h to 4h. In some embodiments, the dosage regimen may comprise administration of a dose over a period of about 1 h.

[0195] In some embodiments, the anti-CTLA4 may be administered over about 30 minutes to over about 3 hours, or over about 1 hour to about 2 hours, or over about one hour.

Anti-PD-1 antibodies; anti-PD-L1 antibodies

[0196] The combination of the disclosure may comprise optionally an anti-PD-1 antibody or an anti-PD-L1 antibody, or a fragment thereof, for the treatment of cancer.

[0197] In one embodiment, the anti-PD-1 antibody or the anti-PD-L1 antibody, or a fragment thereof, is a monoclonal antibody having interfering activity with interaction between PD-1 and PD-L1 . In one embodiment, the anti-PD-1 antibody or the anti-PD-L1 antibody is an IgG.

[0198] Anti-PD-1 antibodies and anti-PD-L1 antibodies capable of interfering with interaction between PD-1 , which is expressed on the surface of immune cells, and PD-L1 , which is expressed on the surface of cancer cells, are useful as immune checkpoint inhibitors, thereby blocking a pathway that shields tumor cells from immune system components able and poised to fight cancer. When PD-1 and PD-L1 interact, they form a biochemical “shield” protecting tumor cells from being destroyed by the immune system. Thus, blockade of either PD-1 or PD-L1 leading to blockade of interaction between PD-1 and PD-L1 prevents or unmasks the biochemical “shield” protecting tumor cells from being destroyed by the immune system.

[0199] A number of anti-PD-1 antibodies have been approved for clinical use in the treatment of cancer. These include pembrolizumab (KEYTRUDA®), nivolumab (OPDIVO®), cemiplimab (LIBTAYO®), sintilimab (TYVYT®), dostarlimab (JEMPERLI®), and tislelizumab.

[0200] Similarly, a number of anti-PD-L1 antibodies have been approved for clinical use in the treatment of cancer. These include atezolizumab (TECENTRIQ®), avelumab (BAVENCIO®), and durvalumab (IMFINZI®).

[0201] In one embodiment, the anti-PD-1 antibody is pembrolizumab or sintilimab.

[0202] In one embodiment, the anti-PD-1 antibody is pembrolizumab. It is a fully human monoclonal lgG1 antibody against human PD-1. [0203] In one embodiment, the anti-PD-1 antibody, or a fragment thereof, comprises the light chain and heavy chain CDRs of pembrolizumab.

[0204] In one embodiment, the anti-PD-1 antibody or a fragment thereof comprises the variable domain of heavy chain (VH) and the variable domain of light chain (VL) of pembrolizumab.

[0205] In one embodiment, the anti-PD-1 antibody is sintilimab. In one embodiment, the anti-PD-1 antibody, or a fragment thereof, comprises the light chain and heavy chain CDRs of sintilimab. In one embodiment, the anti-PD-1 antibody or a fragment thereof comprises the variable domain of heavy chain (VH) and the variable domain of light chain (VL) of sintilimab.

[0206] The anti-PD-1 antibody or the anti-PD-L1 antibody, or a fragment thereof, may also be a single domain antibody or a fragment thereof. In particular, a single domain antibody fragment may consist of a variable heavy chain (VHH) which comprises the CDR1 -H, CDR2- H and CDR3-H of the antibodies as described above. The antibody may also be a heavy chain antibody, i.e., an antibody devoid of light chain, which may or may not contain a CH1 domain.

[0207] The single domain antibody or a fragment thereof may also comprise the framework regions of a camelid single domain antibody, and optionally the constant domain of a camelid single domain antibody.

[0208] The anti-PD-1 antibody or the anti-PD-L1 antibody may also be an antibody fragment, in particular a humanized antibody fragment, selected from the group consisting of Fv, Fab, F(ab')2, Fab', dsFv, (dsFv)2, scFv, sc(Fv)2, and diabodies.

[0209] The antibody may also be a bispecific or multispecific antibody formed from antibody fragments, at least one antibody fragment being an antibody fragment according to the disclosure. The anti-PD-1 antibody or the anti-PD-L1 antibody and fragments thereof can be produced by any technique well known in the art. In particular, said antibodies are produced by techniques as already described.

[0210] The anti-PD-1 antibody or the anti-PD-L1 antibody and fragments thereof can be used in an isolated (e.g., purified) from or contained in a vector, such as a membrane or lipid vesicle (e.g., a liposome).

[0211] The anti-PD-1 antibody or the anti-PD-L1 antibody and fragments thereof may be produced by any technique known in the art, such as, without limitation, any chemical, biological, genetic, or enzymatic technique, either alone or in combination.

Anti-PD-1 antibody or the anti-PD-L1 antibody dosage [0212] In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg. The anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of about 150, 155, 160, 165, 170, 175, 185, 190, 195, 200, 205, 210, 215, 220, 225, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, or about 300 mg.

[0213] In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of about 200 mg.

[0214] According to an embodiment, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, as a loading dose (or first dose).

[0215] According to an embodiment, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered a dose of about 200 mg, as a loading dose (or first dose).

[0216] A loading dose may be administered on day 1 of the first cycle.

[0217] According to an embodiment, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, as a subsequent dose (or second dose).

[0218] According to an embodiment, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered a dose of about 200 mg, as a subsequent dose (or second dose).

[0219] A subsequent dose may be administered on day 1 of cycle(s) subsequent to the first cycle (the subsequent or additional cycles).

[0220] According to an embodiment, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, as a loading dose, on a first cycle of treatment, e.g., at day 1 , and then at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, as a subsequent dose, e.g., at day 1 , on additional cycle(s).

[0221] According to an embodiment, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered a dose of about 200 mg, as a loading dose, on a first cycle of treatment, e.g., at day 1 , and then at a dose of about 200 mg, as a subsequent dose, e.g., at day 1 , on additional cycle(s).

[0222] In some embodiments, the dosage regimen may comprise administration of a dose over a period of about 10 minutes to about 48 hours, or of about 1 h to about 48h, such as over a period of 1 h to 4h. In some embodiments, the dosage regimen may comprise administration of a dose over a period of about 1 h.

[0223] In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered over about 30 minutes to over about 3 hours, or over about 1 hour to about 2 hours, or over about one hour.

Anti-PD1/PDL-1-CTLA4 bispecific antibodies

[0224] In some embodiment, the ADC as disclosed herein may be used with a bispecific antibody directed to PD1/PDL1 and CTLA4 as an alternative of the combined administration of an anti-PD1/PDL1 antibody and an anti-CTLA4 antibody.

[0225] These antibodies simultaneously block the inhibitory signals from both PD1/PDL1 and CTLA4, leading to enhanced T-cell activation and improved anti-tumor responses.

[0226] In some embodiments, a bispecific antibody may be selected from MEDI5752, MDG019, XmAb20717, the bispecific antibodies disclosed in US2019161548, SI-B003 and cadolinimab.

[0227] In some embodiments, a bispecific antibody may be cadolinimab.

[0228] A bispecific antibody may be administered at a dose ranging from about 0.05 mg/kg to about 12 mg/kg.

Combined treatment

Cycle of treatment

[0229] In an embodiment, the treatment duration may be of at least 4 or 6 months.

[0230] A treatment, or a course of treatment, may comprise at least one cycle of treatment.

[0231] In some embodiments, a treatment may comprise a first cycle of treatment, i.e., cycle 1 , and at least one additional, or subsequent, cycle of treatment, i.e., cycle(s) 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, or more.

[0232] In the disclosure, “additional cycle” and “subsequent cycle” are used interchangeably. [0233] In a further embodiment, the antibody-drug conjugate comprising an anti- CEACAM5-antibody, the anti-CTLA4 antibody, and, when presents, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered for a treatment comprising from 1 to 24 cycles, or from 2 to 22 cycles, or from 4 to 20 cycles, or from 6 to 18 cycles, or from 8 to 16 cycles, or from 10 to 14 cycles.

[0234] According to some embodiments, a cycle may last from 1 -week to 10-weeks, or more, or from 1 to 6 weeks, or from 1 to 4 weeks, or from 1 to 3 weeks.

[0235] According to some embodiments, a cycle may be a 1 -week cycle, a 2-weeks cycle, a 3-weeks cycle, a 4-weeks cycle, a 5-weeks cycle, a 6-weeks cycle, a 7-weeks cycle, an 8-weeks cycle, a 9-weeks cycle, or a 10-weeks cycle, or more.

[0236] In some embodiments, a cycle may be a 2-weeks cycle.

[0237] In some embodiments, a cycle may be a 3-weeks cycle.

[0238] In some embodiments, a cycle may be a 4-weeks cycle.

[0239] In some embodiments, the first cycle and the additional cycle(s) may have same or different duration.

[0240] In some embodiments, the first cycle and the additional cycle(s) may have the same duration.

[0241] In some embodiment, a cycle of treatment may comprise a period of treatment on one day or on a plurality of days. The plurality of days may be continuous or may be interrupted by one or a plurality of days of rest, e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10 days of rest.

[0242] In some embodiment, a cycle of treatment may comprise a period of treatment at least on day 1 , for example on day 1 , 2, 3, 4, 5 or 6, of the cycle and a period of rest lasting until the completion of said cycle.

[0243] In some embodiment, a cycle of treatment may comprise a period of treatment on at least a subsequent day relative to day 1 of the cycle, for example on day 2, 3, 4, 5, 6 or 10, of the cycle and a period of rest lasting until the completion of said cycle.

[0244] In some embodiment, a cycle of treatment may comprise a plurality of periods of treatment, for example at least 2, 3, 4 or 5, each separated by a period of rest, and possibly a period of rest lasting until the completion of said cycle.

[0245] The period(s) of treatment and the period(s) of rest may be identical or different between a first cycle and an at least one additional cycle. [0246] In some embodiment, a cycle of treatment, i.e., first and additional cycles, may comprise a period of treatment on day 1 of the cycle and a period of rest lasting until the completion of said cycle.

[0247] In some embodiment, a cycle of treatment, i.e., first and additional cycles, may comprise a period of treatment on a subsequent relative to day 1 of the cycle and a period of rest lasting until the completion of said cycle.

[0248] In some embodiment, a cycle of treatment, i.e., first and additional cycles, may comprise a plurality of periods of treatment, for example at least 2, 3, 4 or 5, each separated by a period of rest, and possibly a period of rest lasting until the completion of said cycle.

[0249] In some embodiments, the periods of treatment and the periods of rest may be identical between a first cycle and an at least one additional cycle.

[0250] In some embodiments, the dose frequency varies from twice a week to once every three weeks, for example every 2 weeks or every 3 weeks.

[0251] In some embodiments, a dose frequency may be once every three weeks.

[0252] In some embodiments, a dose frequency may be once every two weeks.

[0253] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody, the anti-CTLA4 antibody, and, when presents, the anti-PD-1 antibody or anti-PD-L1 antibody may be administered on the same day, or each at different days of a cycle of treatment.

[0254] In some embodiments, one of the antibody-drug conjugate comprising an anti- CEACAM5-antibody, the anti-CTLA4 antibody, and, when presents, the anti-PD-1 antibody or anti-PD-L1 antibody may be administered on day 1 of a cycle of treatment, and the two others may be administered on one same subsequent day or on subsequent separate days, of the cycle of treatment.

[0255] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody, the anti-CTLA4 antibody, and, when presents, the anti-PD-1 antibody or anti-PD-L1 antibody may be administered on day 1 of a first cycle of treatment and on day 1 of an at least one additional cycle(s) of treatment. The antibody-drug conjugate comprising an anti-CEACAM5-antibody, the anti-CTLA4 antibody, and, when presents, the anti-PD-1 antibody or anti-PD-L1 antibody may be administered on day 1 of each cycle of treatment.

[0256] In some embodiments, the ADC may be administered on day 1 of a first cycle of treatment and of at least one subsequent cycle of treatment, and the anti-CTLA4 antibody, and, when presents, the anti-PD-1 antibody or anti-PD-L1 antibody may be administered on a subsequent day of a first cycle of treatment and of at least one subsequent cycle of treatment.

[0257] In some embodiments, the anti-CTLA4 antibody, and, when presents, the anti- PD-1 antibody or anti-PD-L1 antibody may be administered on at least one subsequent day, for example on 1 to 20, or 2 to 18, or 4 to 16, or 6 to 14 or 8 to 12 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by one or a plurality, e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10 days of rest.

[0258] In some embodiments, the anti-CTLA4 antibody, and, when presents, the anti- PD-1 antibody or anti-PD-L1 antibody may be administered on at least one subsequent day, for example on at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by one or a plurality, e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10 days of rest.

[0259] In some embodiments, the anti-CTLA4 antibody, and, when presents, the anti- PD-1 antibody or anti-PD-L1 antibody may be administered on at least one subsequent day, for example on at least 1 , 2, 3, 4, 5, or 6 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by one or a plurality, e.g., 2, 3, 4, 5, or 6, days of rest.

[0260] In some embodiments, the anti-CTLA4 antibody, and, when presents, the anti- PD-1 antibody or anti-PD-L1 antibody may be administered on at least one subsequent day, for example on at least 4 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by one or a plurality, e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10 days of rest.

[0261] In some embodiments, a cycle may last three weeks, and the ADC may be administered on day 1 of a first cycle of treatment and of at least one subsequent cycle of treatment, and the anti-CTLA4 antibody, and, when presents, the anti-PD-1 antibody or anti- PD-L1 antibody may be administered on subsequent day 10, 13, 17 and 20 of a first cycle of treatment and of at least one subsequent cycle of treatment. The anti-CTLA4 antibody, and, when presents, the anti-PD-1 antibody or anti-PD-L1 antibody may be administered sequentially, separately, or simultaneously. The anti-CTLA4 antibody, and, when presents, the anti-PD-1 antibody or anti-PD-L1 antibody may be administered simultaneously.

[0262] In some embodiments, the first cycle and the additional cycle(s) may comprise the administration of same or different dosage for the ADC and/or the anti-CTLA4 antibody, and/or, when presents, the anti-PD-1 antibody or anti-PD-L1 antibody. [0263] The dosage of the ADC and/or the anti-CTLA4 antibody, and/or, when presents, the anti-PD-1 antibody or anti-PD-L1 antibody used for a first cycle may be different than the dosage for an additional cycle. A dosage for a first cycle may be a "loading” dose. A dosage for a subsequent cycle may be a “subsequent” dose.

[0264] For example, the first cycle may comprise an administration of a loading dose (or first dose), and the additional cycle(s) may comprise an administration of a subsequent dose (or second), i.e., different dosages for the loading and subsequent doses.

[0265] In some embodiments, a loading dose may be higher than a subsequent dose.

[0266] Alternatively, the first and additional cycles may comprise an administration of a same dose, i.e., same dosage for the loading and subsequent doses. In such embodiment, “loading” and “subsequent” doses are named “dose”.

[0267] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered at a loading dose in a first cycle and at a subsequent dose in additional cycle(s), i.e., different dosages for the loading and subsequent doses.

[0268] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered at a same dose in a first cycle and additional cycle(s), i.e., same dosage for the loading and subsequent doses.

[0269] In some embodiments, the anti-CTLA4 antibody may be administered at a loading dose in a first cycle and at a subsequent dose in additional cycle(s), i.e., different dosages for the loading and subsequent doses.

[0270] In some embodiments, the anti-CTLA4 antibody may be administered at a same dose in a first cycle and additional cycle(s), i.e., same dosage for the loading and subsequent doses.

[0271 ] In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a loading dose in a first cycle and at a subsequent dose in additional cycle(s), i.e., different dosages for the loading and subsequent doses.

[0272] In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a same dose in a first cycle and additional cycle(s), i.e., same dosage for the loading and subsequent doses.

[0273] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered at a same dose in a first cycle and additional cycle(s), the anti-CTLA4 antibody, and, when presents, the anti-PD-1 antibody or the anti-PD- L1 antibody may be administered at a same dose in a first cycle and additional cycle(s), i.e., same dosage for the loading and subsequent doses.

[0274] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered at a loading dose in a first cycle and at a subsequent dose in additional cycle(s), i.e., different dosages for the loading and subsequent doses, the anti-CTLA4 antibody may be administered at a same dose in a first cycle and additional cycle(s), i.e., same dosage for the loading and subsequent doses, and, when presents, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a same dose in a first cycle and additional cycle(s), i.e., same dosage for the loading and subsequent doses.

[0275] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered at a same dose in a first cycle and additional cycle(s), the anti-CTLA4 antibody may be administered at a loading dose in a first cycle and at a subsequent dose in additional cycle(s), i.e., different dosages for the loading and subsequent doses, and, when presents, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a same dose in a first cycle and additional cycle(s), i.e., same dosage for the loading and subsequent doses.

[0276] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered at a loading dose in a first cycle and at a subsequent dose in additional cycle(s), i.e., different dosages for the loading and subsequent doses, the anti-CTLA4 antibody may be administered at a loading dose in a first cycle and at a subsequent dose in additional cycle(s), i.e., different dosages for the loading and subsequent doses, and, when presents, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a same dose in a first cycle and additional cycle(s), i.e., same dosage for the loading and subsequent doses.

[0277] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered at a loading dose in a first cycle and at a subsequent dose in additional cycle(s), i.e., different dosages for the loading and subsequent doses, the anti-CTLA4 antibody may be administered at a loading dose in a first cycle and at a subsequent dose in additional cycle(s), i.e., different dosages for the loading and subsequent doses, and, when presents, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a same dose in a first cycle and additional cycle(s), i.e., different dosage for the loading and subsequent doses.

[0278] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered at a loading dose in a first cycle and at a subsequent dose in additional cycle(s), i.e., different dosages for the loading and subsequent doses, the anti-CTLA4 antibody may be administered at a loading dose in a first cycle and at a subsequent dose in additional cycle(s), i.e., same dosages for the loading and subsequent doses, and, when presents, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a same dose in a first cycle and additional cycle(s), i.e., different dosage for the loading and subsequent doses.

Sequences of administration

[0279] The disclosure relates to an antibody-drug conjugate comprising an anti- CEACAM5-antibody for use for treating cancer in a subject in need thereof who receives, simultaneously, separately, or sequentially an anti-CTLA4 antibody. Optionally, an anti-PD-1 antibody or anti-PD-L1 antibody may be administered simultaneously, separately, or sequentially with the ADC and the anti-CTLA4 antibody.

[0280] According to the present disclosure, the ADC is for use for treating cancer in combination with an anti-CTLA4 antibody and, optionally, an anti-PD-1 antibody or anti-PD-L1 antibody. The disclosure also relates to an anti-CTLA4 antibody for use for treating cancer in combination with the ADC and, optionally, an anti-PD-1 antibody or anti-PD-L1 antibody. The disclosure also relates to an anti-PD-1 antibody or anti-PD-L1 antibody for use for treating cancer in combination with the ADC and an anti-CTLA4 antibody.

[0281] In some embodiments, (i) the antibody-drug conjugate, (ii) the anti-CTLA4 antibody, and, optionally, (iii) the anti-PD-1 antibody or anti-PD-L1 antibody may be administered simultaneously (or concurrently), separately or sequentially to a subject in need thereof.

[0282] As used herein, the expression “in combination with” means that the anti- CTLA4 antibody or the anti-PD-1 antibody or anti-PD-L1 antibody are administered before, after, or concurrent with the ADC. In some embodiments, the term “in combination with” includes sequential or concomitant administration of the ADC and the anti-CTLA4 antibody or the anti-PD-1 antibody or anti-PD-L1 antibody. In some embodiments, the term “combination" used with respect to the anti-CTLA4 antibody, the antibody-drug conjugate comprising an anti- CEACAM5-antibody, and the anti-PD-1 antibody or the anti-PD-L1 antibody intends to refer to antibodies formulated individually and administered sequentially to a subject in need thereof, for a cancer indication.

[0283] Methods to treat cancer (e.g., colorectal, GC, GEJ cancer, NSQ NSCLC) includes administering an antibody-drug conjugate comprising an anti-CEACAM5-antibody (e.g., tusamitamab ravtansine) in combination with an anti-CTLA4 antibody and, optionally, an anti-PD-1 antibody or anti-PD-L1 antibody for additive or synergistic activity.

[0284] Methods to treat cancer (e.g., colorectal, GC, GEJ cancer, NSQ NSCLC) includes administering an antibody-drug conjugate comprising an anti-CEACAM5-antibody (e.g., tusamitamab ravtansine) in combination with an anti-CTLA4 antibody and, optionally, an anti-PD-1 antibody or anti-PD-L1 antibody for synergistic activity.

[0285] In some embodiments, (i) the antibody-drug conjugate and (ii) the anti-CTLA4 antibody are administered sequentially to a subject in need thereof. The anti-PD-1 antibody or anti-PD-L1 antibody, when presents, is administered, relative to the ADC, sequentially to a subject in need thereof. The CTLA4 antibody and, when presents, the anti-PD-1 antibody or anti-PD-L1 antibody may be administered sequentially or simultaneously. The CTLA4 antibody and the anti-PD-1 antibody or anti-PD-L1 antibody may be administered simultaneously.

[0286] According to some embodiments, the ADC and the anti-CTLA4 antibody are administered simultaneously, separately, or sequentially to a subject in need thereof.

[0287] According to an embodiment, the ADC and the anti-CTLA4 antibody are simultaneously administered to a subject in need thereof. For example, the ADC and the anti- CTLA4 antibody are administered on day one of a cycle of treatment, approximatively at the same time. A simultaneous administration of the ADC and the anti-CTLA4 antibody may be administered by the same route.

[0288] According to an embodiment, the ADC and the anti-CTLA4 antibody are separately administered to a subject in need thereof. For example, the ADC and the anti- CTLA4 antibody are administered on day one of a cycle, by separate routes or at separates location of the body of said subject. A separate administration of the ADC and the anti-CTLA4 antibody may be at the same time or at close times, e.g., 5 min or less.

[0289] According to an embodiment, the ADC and the anti-CTLA4 antibody are sequentially administered to a subject in need thereof. For example, the ADC and the anti- CTLA4 antibody are administered on day one of a cycle, at different times, for example the ADC is administered one to three hours before or after the anti-CTLA4 antibody. [0290] The ADC may be administered about 0.5 hr, 1 hr, 2hrs, 3hrs, 4hrs, 5hrs or about 6hrs before or after the anti-CTLA4 antibody. The ADC may be administered about 1 hr before or after the anti-CTLA4 antibody.

[0291] In a sequential administration, the period of time between the administration of the anti-CTLA4 antibody and the antibody-drug conjugate comprising an anti-CEACAM5- antibody may last from about a few minutes to about several hours, days, or weeks. In some embodiments, the period of time may range from about 5 minutes to about 5 hours, from about 5 minutes to about 3 hours, for example from 10 minutes to about 2.5 hours, from about 30 minutes to about 2 hours, or from about 1 hour to about 1 .5 hours. A period of time between the administration of the anti-CTLA4 antibody and the antibody-drug conjugate comprising an anti-CEACAM5-antibody may last about 5 minutes, about 10 minutes, about 30 minutes, about 1 hour, about 1 .5 hours, about 2 hours, about 2.5 hours or about 3 hours.

[0292] The period of time between an administration of the anti-CTLA4 antibody and an administration of the ADC may be about 72 hours, about 60 hours, about 48 hours, about 36 hours, about 24 hours, about 12 hours, about 10 hours, about 8 hours, about 6 hours, about 4 hours, about 2 hours, about 1 hour, about 30 minutes, about 15 minutes, or about 10 minutes.

[0293] The period of time between an administration of the anti-CTLA4 antibody and an administration of the antibody-drug conjugate comprising an anti-CEACAM5-antibody may range from about 20 minutes to about 5 hours, from about 30 minutes to about 3 hours, from about 40 minutes to about 2 hours, from about 50 minutes to about 1.5 hours, or may last about 1 hour.

[0294] In an embodiment, in a sequential administration, the period of time between the administration of the anti-CTLA4 antibody and the antibody-drug conjugate comprising an anti-CEACAM5-antibody may be at least one hour.

[0295] In some embodiments, the ADC and the anti-CTLA4 antibody are administered on different days of a cycle. For example, the ADC is administered on day one of a cycle and the CTLA4 antibody is administered on a subsequent day of a cycle, for example on day 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 13, 14, 15, 16, 17, 18, 19, 20, or 21 , or reciprocally.

[0296] A sequential administration of the ADC and the anti-CTLA4 antibody may be administered by separate routes or by a same route.

[0297] A sequential administration of the ADC and the anti-CTLA4 antibody may comprise administration of the ADC before or after the anti-CTLA4 antibody. [0298] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered before the anti-CTLA4 antibody.

[0299] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered after the anti-CTLA4 antibody.

[0300] In some embodiments, the sequence of administration of the antibody-drug conjugate comprising an anti-CEACAM5-antibody and of the anti-CTLA4 antibody may vary along the cycles of treatment. In some embodiments, one or more cycles of a treatment may comprise a first sequence of administration and one or more cycles of said treatment may comprise a second sequence of administration, the first and second sequences being different.

[0301] In some embodiments, the sequence of administration of the antibody-drug conjugate comprising an anti-CEACAM5-antibody and of the anti-CTLA4 antibody may be the same for all cycles of treatment.

[0302] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody and the anti-CTLA4 antibody may be administered on the same day, or each at different days of a cycle of treatment.

[0303] In some embodiments, one of the antibody-drug conjugate comprising an anti- CEACAM5-antibody and of the anti-CTLA4 antibody may be administered on day 1 of a cycle of treatment, and the other may be administered on a subsequent day of the cycle of treatment.

[0304] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered on day 1 of a first cycle of treatment. In some embodiments, the antibody-drug conjugate comprising an anti-CEACAM5-antibody may be administered on day 1 of an at least one additional cycle(s) of treatment. In some embodiments, the anti-CTLA4 antibody may be administered on day 1 of a first cycle of treatment. In some embodiments, the anti-CTLA4 antibody may be administered on day 1 of an at least one additional cycle(s) of treatment.

[0305] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody and the anti-CTLA4 antibody may be administered on day 1 of a first cycle of treatment. In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody and the anti-CTLA4 antibody may be administered on day 1 of an at least one additional cycle(s) of treatment.

[0306] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody and the anti-CTLA4 antibody may be administered on day 1 of a first cycle of treatment and on day 1 of an at least one additional cycle(s) of treatment. The antibody- drug conjugate comprising an anti-CEACAM5-antibody and the anti-CTLA4 antibody may be administered on day 1 of each cycle of treatment.

[0307] In some embodiments, the ADC may be administered on day 1 of a first cycle of treatment, and the anti-CTLA4 antibody may be administered on a subsequent day of a first cycle of treatment. In some embodiments, the ADC may be administered on day 1 of at least one subsequent cycle of treatment, and the anti-CTLA4 antibody may be administered on a subsequent day of at least one subsequent cycle of treatment.

[0308] In some embodiments, the ADC may be administered on day 1 of a first cycle of treatment and of at least one subsequent cycle of treatment, and the anti-CTLA4 antibody may be administered on a subsequent day of a first cycle of treatment and of at least one subsequent cycle of treatment.

[0309] In some embodiments, the anti-CTLA4 antibody may be administered on at least one subsequent day, for example on 1 to 20, or 2 to 18, or 4 to 16, or 6 to 14 or 8 to 12 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by one or a plurality of days of rest.

[0310] In some embodiments, the anti-CTLA4 antibody may be administered on at least one subsequent day, for example on at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by one or a plurality of days of rest.

[0311] In some embodiments, the anti-CTLA4 antibody may be administered on at least one subsequent day, for example on at least 1 , 2, 3, 4, 5, or 6 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by 1 , 2, 3, 4, 6, or 7 of days of rest.

[0312] In some embodiments, the anti-CTLA4 antibody may be administered on at least one subsequent day, for example on at least 4 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The subsequent days may be continuous or interrupted by 1 , 2, 3, 4, 6, or 7 of days of rest.

[0313] In some embodiments, a cycle may last three weeks, and the ADC may be administered on day 1 of a first cycle of treatment and of at least one subsequent cycle of treatment, and the anti-CTLA4 antibody may be administered on day 10, 13, 17 and 20 of a first cycle of treatment and of at least one subsequent cycle of treatment. [0314] According to some embodiments, an anti-PD-1 antibody or an anti-PD-L1 antibody may be administered with the combination comprising an ADC and an anti-CTLA4 antibody as disclosed herein.

[0315] According to some embodiments, (i) the ADC, (ii) the anti-CTLA4 antibody, and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered simultaneously, separately, or sequentially to a subject in need thereof.

[0316] According to an embodiment, (i) the ADC, (ii) the anti-CTLA4 antibody, and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody may be simultaneously administered to a subject in need thereof. For example, (i) ADC, (ii) the anti-CTLA4 antibody, and (iii) the anti- PD-1 antibody or the anti-PD-L1 antibody may be administered on day one of a cycle of treatment, approximatively at the same time. A simultaneous administration of the ADC, the anti-CTLA4 antibody, and the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered by the same route.

[0317] According to an embodiment, the (i) ADC, (ii) the anti-CTLA4 antibody, and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody are separately administered to a subject in need thereof. For example, the (i) ADC, the (ii) anti-CTLA4 antibody, and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody are administered on day one of a cycle, by separate routes or at separates location of the body of said subject. A separate administration of the ADC, the anti-CTLA4 antibody, and the anti-PD-1 antibody or the anti-PD-L1 antibody may be at the same time or at close times, e.g., 5 min or less.

[0318] According to an embodiment, the (i) ADC, the (ii) anti-CTLA4 antibody, and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody are sequentially administered to a subject in need thereof. For example, the (i) ADC, the (ii) anti-CTLA4 antibody, (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody are administered on day one of a cycle, at different times, for example the ADC is administered one to three hours after the anti-CTLA4 antibody and/or the anti-PD-1 antibody or the anti-PD-L1 antibody.

[0319] In some embodiments, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody are simultaneously, separately or sequentially administered.

[0320] In some embodiments, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody are simultaneously administered as a combination. [0321] In some embodiments, the ADC is administered sequentially to a simultaneous administration of the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody.

[0322] The ADC may be administered about 0.5 hr, 1 hr, 2hrs, 3hrs, 4hrs, 5hrs or about 6hrs before or after the anti-CTLA4 antibody and/or before or after the anti-PD-1 antibody or the anti-PD-L1 antibody, or a simultaneous administration of the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody. The ADC may be administered about 1 hr before or after the anti-CTLA4 antibody and/or before or after the anti-PD-1 antibody or the anti-PD- L1 antibody and/or before or after a simultaneous administration of the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody.

[0323] In a sequential administration, the period of time between the administration of the antibody-drug conjugate comprising an anti-CEACAM5-antibody, the administration of the anti-CTLA4 antibody, and the administration of the anti-PD-1 antibody or the anti-PD-L1 antibody, or a simultaneous administration of the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, may last from about a few minutes to about several hours, days, or weeks. In some embodiments, the period of time may range from about 5 minutes to about 5 hours, from about 5 minutes to about 3 hours, for example from 10 minutes to about 2.5 hours, from about 30 minutes to about 2 hours, or from about 1 hour to about 1 .5 hours. A period of time between the administration of the antibody-drug conjugate comprising an anti- CEACAM5-antibody, the administration of the anti-CTLA4 antibody, and the administration of the anti-PD-1 antibody or the anti-PD-L1 antibody, or a simultaneous administration of the anti- CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody may last about 5 minutes, about 10 minutes, about 30 minutes, about 1 hour, about 1.5 hours, about 2 hours, about 2.5 hours or about 3 hours.

[0324] The period of time between an administration of the ADC, an administration of the anti-CTLA4 antibody, and an administration of the anti-PD-1 antibody or the anti-PD-L1 antibody, or a simultaneous administration of the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, may be about 72 hours, about 60 hours, about 48 hours, about 36 hours, about 24 hours, about 12 hours, about 10 hours, about 8 hours, about 6 hours, about 4 hours, about 2 hours, about 1 hour, about 30 minutes, about 15 minutes, or about 10 minutes.

[0325] The period of time between an administration of the antibody-drug conjugate comprising an anti-CEACAM5-antibody, an administration of the anti-CTLA4 antibody, and an administration of the anti-PD-1 antibody or the anti-PD-L1 antibody, or a simultaneous administration of the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, may range from about 20 minutes to about 5 hours, from about 30 minutes to about 3 hours, from about 40 minutes to about 2 hours, from about 50 minutes to about 1 .5 hours, or may last about 1 hour.

[0326] In an embodiment, in a sequential administration, the period of time between the administration of the antibody-drug conjugate comprising an anti-CEACAM5-antibody, of the anti-CTLA4 antibody, and of the anti-PD-1 antibody or the anti-PD-L1 antibody, or a simultaneous administration of the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti- PD-L1 antibody, may be at least one hour.

[0327] In some embodiments, the (i) ADC, the (ii) anti-CTLA4 antibody and (iii) the anti- PD-1 antibody or the anti-PD-L1 antibody, or a combination of the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, are administered on different days of a cycle. For example, the ADC is administered on day one of a cycle and the CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or a combination of the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, are administered on at least one subsequent day of a cycle, for example on day 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 13, 14, 15, 16, 17, 18, 19, 20, or 21 , or reciprocally.

[0328] A sequential administration of the ADC, the anti-CTLA4 antibody and the anti- PD-1 antibody or the anti-PD-L1 antibody, or a combination of the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered by separate routes or by a same route.

[0329] A sequential administration of the ADC, the anti-CTLA4 antibody and the anti- PD-1 antibody or the anti-PD-L1 antibody, or a combination of the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, may comprise administration of the ADC before or after the anti-CTLA4 antibody, before or after the anti-PD-1 antibody or the anti-PD- L1 antibody, or before or after the combination of the anti-CTLA4 antibody and of the anti-PD- 1 antibody or the anti-PD-L1 antibody.

[0330] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered before the anti-CTLA4 antibody, the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody.

[0331] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered after the anti-CTLA4 antibody, the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody. [0332] In some embodiments, the anti-CTLA4 antibody may be administered after, before or simultaneously with the anti-PD-1 antibody or the anti-PD-L1 antibody.

[0333] In some embodiments, the sequence of administration of the antibody-drug conjugate comprising an anti-CEACAM5-antibody, of the anti-CTLA4 antibody and of the anti- PD-1 antibody or the anti-PD-L1 antibody, or of the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may vary along the cycles of treatment. In some embodiments, one or more cycles of a treatment may comprise a first sequence of administration and one or more cycles of said treatment may comprise a second sequence of administration, the first and second sequences being different.

[0334] In some embodiments, the sequence of administration of the antibody-drug conjugate comprising an anti-CEACAM5-antibody, of the anti-CTLA4 antibody and of the anti- PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be the same for all cycles of treatment.

[0335] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on the same day, or each at different days of a cycle of treatment.

[0336] In some embodiments, one of the antibody-drug conjugate comprising an anti- CEACAM5-antibody, of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD- L1 antibody, or of the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on day 1 of a cycle of treatment, and the other(s) may be administered on a subsequent day of the cycle of treatment.

[0337] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered on day 1 of a first cycle of treatment. In some embodiments, the anti-CTLA4 antibody may be administered on day 1 of a first cycle of treatment. In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered on day 1 of a first cycle of treatment. In some embodiments, the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered on day 1 of a first cycle of treatment. In some embodiments, the antibody-drug conjugate comprising an anti-CEACAM5-antibody may be administered on day 1 of an at least one additional cycle(s) of treatment. In some embodiments, the anti-CTLA4 antibody may be administered on day 1 of an at least one additional cycle(s) of treatment. In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered on day 1 of an at least one additional cycle(s) of treatment. In some embodiments, the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered on day 1 of an at least one additional cycle(s) of treatment. In some embodiments, the antibody-drug conjugate comprising an anti-CEACAM5-antibody, the anti- CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on day 1 of an at least one additional cycle(s) of treatment.

[0338] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on day 1 of a first cycle of treatment. In some embodiments, the antibody-drug conjugate comprising an anti-CEACAM5-antibody, the anti- CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on day 1 of an at least one additional cycle(s) of treatment.

[0339] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on day 1 of a first cycle of treatment and on day 1 of an at least one additional cycle(s) of treatment. The antibody-drug conjugate comprising an anti-CEACAM5-antibody, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD- L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on day 1 of each cycle of treatment.

[0340] In some embodiments, the ADC may be administered on day 1 of a first cycle of treatment, and the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on a subsequent day of a first cycle of treatment. In some embodiments, the ADC may be administered on day 1 of at least one subsequent cycle of treatment, and the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on a subsequent day of at least one subsequent cycle of treatment. [0341] In some embodiments, the ADC may be administered on day 1 of a first cycle of treatment and of at least one subsequent cycle of treatment, and the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on a subsequent day of a first cycle of treatment and of at least one subsequent cycle of treatment.

[0342] In some embodiments, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on at least one subsequent day, for example on 1 to 20, or 2 to 18, or 4 to 16, or 6 to 14 or 8 to 12 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by one or a plurality of days of rest.

[0343] In some embodiments, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on at least one subsequent day, for example on at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by one or a plurality of days of rest.

[0344] In some embodiments, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on at least one subsequent day, for example on at least 1 , 2, 3, 4, 5, or 6 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by 1 , 2, 3, 4, 6, or 7 of days of rest.

[0345] In some embodiments, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on at least one subsequent day, for example on at least 4 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The subsequent days may be continuous or interrupted by 1 , 2, 3, 4, 6, or 7 of days of rest.

[0346] In some embodiments, a cycle may last three weeks, and the ADC may be administered on day 1 of a first cycle of treatment and of at least one subsequent cycle of treatment, and the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on day 10, 13, 17 and 20 of a first cycle of treatment and of at least one subsequent cycle of treatment.

[0347] In some embodiments, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody may be simultaneously administered as a combination.

[0348] The present disclosure also relates to a method of treatment of cancer in a subject in need thereof, comprising administering an antibody-drug conjugate comprising an anti-CEACAM5-antibody and administering an anti-CTLA4 antibody to the subject in need thereof.

[0349] The present disclosure also relates to a method of treatment of cancer in a subject in need thereof, comprising administering, sequentially, in the following order: (a) an antibody-drug conjugate comprising an anti-CEACAM5-antibody and (b) an anti-CTLA4 antibody to the subject in need thereof.

[0350] The present disclosure also relates to a method of treatment of cancer in a subject in need thereof, comprising administering, sequentially, in the following order: (a) an anti-CTLA4 antibody and (b) an antibody-drug conjugate comprising an anti-CEACAM5- antibody to the subject in need thereof.

[0351] The present disclosure also relates to a method of treatment of cancer in a subject in need thereof, comprising administering an antibody-drug conjugate comprising an anti-CEACAM5-antibody, administering an anti-CTLA4 antibody, and administering an anti- PD-1 antibody or an anti-PD-L1 antibody, or administering a combination of an anti-CTLA4 antibody and of an anti-PD-1 antibody or an anti-PD-L1 antibody, to the subject in need thereof.

[0352] The present disclosure also relates to a method of treatment of cancer in a subject in need thereof, comprising administering, sequentially, in the following order: (a) an antibody-drug conjugate comprising an anti-CEACAM5-antibody, (b) an anti-CTLA4 antibody, and (c) an anti-PD-1 antibody or an anti-PD-L1 antibody to the subject in need thereof.

[0353] The present disclosure also relates to a method of treatment of cancer in a subject in need thereof, comprising administering, sequentially, in the following order: (a) an antibody-drug conjugate comprising an anti-CEACAM5-antibody, (b) an anti-PD-1 antibody or an anti-PD-L1 antibody, and (c) an anti-CTLA4 antibody to the subject in need thereof.

[0354] The present disclosure also relates to a method of treatment of cancer in a subject in need thereof, comprising administering, sequentially, in the following order: (a) an antibody-drug conjugate comprising an anti-CEACAM5-antibody and (b) a combination of an anti-CTLA4 antibody and of an anti-PD-1 antibody or an anti-PD-L1 antibody to the subject in need thereof.

[0355] The present disclosure also relates to a method of treatment of cancer in a subject in need thereof, comprising administering, sequentially, in the following order: (a) an anti-CTLA4 antibody, (b) an antibody-drug conjugate comprising an anti-CEACAM5-antibody, and (c) an anti-PD-1 antibody or an anti-PD-L1 antibody to the subject in need thereof.

[0356] The present disclosure also relates to a method of treatment of cancer in a subject in need thereof, comprising administering, sequentially, in the following order: (a) an anti-CTLA4 antibody, (b) an anti-PD-1 antibody or an anti-PD-L1 antibody, and (c) an antibodydrug conjugate comprising an anti-CEACAM5-antibody to the subject in need thereof.

[0357] The present disclosure also relates to a method of treatment of cancer in a subject in need thereof, comprising administering, sequentially, in the following order: (a) an anti-PD-1 antibody or an anti-PD-L1 antibody, (b) an anti-CTLA4 antibody, and (c) an antibodydrug conjugate comprising an anti-CEACAM5-antibody to the subject in need thereof.

[0358] The present disclosure also relates to a method of treatment of cancer in a subject in need thereof, comprising administering, sequentially, in the following order: (a) a combination of an anti-CTLA4 antibody and of an anti-PD-1 antibody or an anti-PD-L1 antibody and (b) an antibody-drug conjugate comprising an anti-CEACAM5-antibody to the subject in need thereof.

[0359] The present disclosure also relates to a combination comprising an antibodydrug conjugate comprising an anti-CEACAM5-antibody, an anti-CTLA4 antibody, and an anti- PD-1 antibody or an anti-PD-L1 antibody for use for treating cancer.

[0360] The combination is for a sequential administration of the antibody-drug conjugate comprising an anti-CEACAM5-antibody, the anti-CTLA4 antibody, and the anti-PD- 1 antibody or the anti-PD-L1 antibody.

[0361] A method of treatment or a use, as disclosed herein, may achieve a synergistic effect in reducing tumor size.

[0362] A method of treatment or a use, as disclosed herein, may achieve a synergistic effect in inhibiting tumor growth.

Combination dosages [0363] In some embodiments, (i) the antibody-drug conjugate may be administered at a dose of 80 mg/m² to 210 mg/m², 80 mg/m² to 170 mg/m², or at a dose of 80 mg/m² to 150 mg/m², or at a dose of 80 mg/m² to 120 mg/m², or at a dose of 80 mg/m² to 100 mg/m², and (ii) the anti-CTLA4 antibody may be administered at a dose of 1 mg/kg to 10 mg/kg, or at a dose of 3 mg/k to 10 mg/kg.

[0364] In some embodiments, (i) the antibody-drug conjugate may be administered at a dose level of 80, 100, 120, 150, 170, 180, or 210 mg/m² and (ii) the anti-CTLA4 antibody may be administered at a dose of 1 mg/kg, or at a dose of 2.5 mg/kg, or at a dose of 3 mg/kg, or at a dose of 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of 10 mg/kg.

[0365] In some embodiments, (i) the antibody-drug conjugate may be administered at a dose of 80 mg/m² to 210 mg/m², 80 mg/m² to 170 mg/m², or at a dose of 80 mg/m² to 150 mg/m², or at a dose of 80 mg/m² to 120 mg/m², or at a dose of 80 mg/m² to 100 mg/m², (ii) the anti-CTLA4 antibody may be administered at a dose of 1 mg/kg to 10 mg/kg, or at a dose of 3 mg/k to 10 mg/kg, and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of 150 mg to 400 mg, or at a dose of 150 mg to 300 mg.

[0366] In some embodiments, (i) the antibody-drug conjugate may be administered at a dose level of 80, 100, 120, 150, 170, 180, or 210 mg/m² (ii) the anti-CTLA4 antibody may be administered at a dose of 1 mg/kg, or at a dose of 2.5 mg/kg, or at a dose of 3 mg/kg, or at a dose of 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of 10 mg/kg, and (iii) the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of 200 mg.

[0367] In some embodiments, the dosage regimen may comprise administration of a dose over a period of about 10 minutes to about 48 hours, or of about 1 h to about 48h, such as over a period of 1 h to 4h. In some embodiments, the dosage regimen may comprise administration of a dose over a period of about 1 h.

[0368] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m² of surface body area of a subject in need thereof on day 1 of a first cycle of treatment. In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m² on day 1 of an at least one additional cycle(s) of treatment. In some embodiments, the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg of weight of a subject in need thereof on day 1 of a first cycle of treatment. In some embodiments, the anti- CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg on day 1 of an at least one additional cycle(s) of treatment.

[0369] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m² and the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, on day 1 of a first cycle of treatment, as a loading dose.

[0370] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m² and the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, on day 1 of at least one additional cycle(s) of treatment, as a subsequent dose.

[0371] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m² and the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, on day 1 of a first cycle of treatment, as a loading dose, and the antibody-drug conjugate comprising the anti-CEACAM5 antibody may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m² and the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, on day 1 of at least one additional cycle(s) of treatment, as a subsequent dose.

[0372] In some embodiments the loading dose and the subsequent dose are the same.

[0373] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m² on day 1 of a first cycle of treatment, as a loading dose, and the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, on a subsequent day of a first cycle of treatment, as a loading dose.

[0374] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m² on day 1 of at least one additional cycle(s) of treatment, as a subsequent dose, and the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, on a subsequent day of at least one additional cycle(s) of treatment, as a subsequent dose.

[0375] In some embodiments, the ADC may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m² on day 1 of a first cycle of treatment and of at least one subsequent cycle of treatment, and the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg on a subsequent day of a first cycle of treatment and of at least one subsequent cycle of treatment.

[0376] In some embodiments, the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg on at least one subsequent day, for example on 1 to 20, or 2 to 18, or 4 to 16, or 6 to 14 or 8 to 12 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by one or a plurality of days of rest.

[0377] In some embodiments, the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg on at least one subsequent day, for example on at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by one or a plurality of days of rest.

[0378] In some embodiments, the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg on at least one subsequent day, for example on at least 1 , 2, 3, 4, 5, or 6 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by 1 , 2, 3, 4, 6, or 7 of days of rest.

[0379] In some embodiments, the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg on at least one subsequent day, for example on at least 4 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The subsequent days may be continuous or interrupted by 1 , 2, 3, 4, 6, or 7 of days of rest.

[0380] In some embodiments, a cycle may last three weeks, and the ADC may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m² on day 1 of a first cycle of treatment and of at least one subsequent cycle of treatment, and the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 4 mg/kg, or at a dose of about 10 mg/kg on day 10, 13, 17 and 20 of a first cycle of treatment and of at least one subsequent cycle of treatment.

[0381] In some embodiments, the antibody-drug conjugate comprising an anti- CEACAM5-antibody may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m² on day 1 of a first cycle of treatment. In some embodiments, the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg on day 1 of a first cycle of treatment. In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg on day 1 of a first cycle of treatment. In some embodiments, the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered, respectively, at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg and at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg on day 1 of a first cycle of treatment. In some embodiments, the antibody-drug conjugate comprising an anti-CEACAM5-antibody may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m²on day 1 of an at least one additional cycle(s) of treatment. In some embodiments, the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg on day 1 of an at least one additional cycle(s) of treatment. In some embodiments, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg on day 1 of an at least one additional cycle(s) of treatment. In some embodiments, the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered, respectively, at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg on day 1 and at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg of an at least one additional cycle(s) of treatment. In some embodiments, the antibodydrug conjugate comprising an anti-CEACAM5-antibody, the anti-CTLA4 antibody and the anti- PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered, respectively, at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m², on day 1 of an at least one additional cycle(s) of treatment.

[0382] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m², the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, and the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, on day 1 of a first cycle of treatment, as a loading dose.

[0383] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m², the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, and the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, on day 1 of at least on additional cycle(s) of treatment, as a subsequent dose. [0384] In some embodiments, the antibody-drug conjugate comprising (i) an anti- CEACAM5-antibody, (ii) the anti-CTLA4 antibody and (iii) the anti-PD-1 antibody or the anti- PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on day 1 of a first cycle of treatment and on day 1 of an at least one additional cycle(s) of treatment, respectively, (i) at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m, (ii) at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, and (iii) at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg. The (i) antibody-drug conjugate comprising an anti-CEACAM5- antibody, the (ii) anti-CTLA4 antibody and the (iii) anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered on day 1 of each cycle of treatment, respectively, (i) at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m, (ii) at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, and (iii) at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg.

[0385] In some embodiments the loading dose and the subsequent dose are the same.

[0386] A cycle may be 2 or 3 weeks.

[0387] The ADC may be administered before the anti-CTLA4 antibody, and, when presents, the anti-PD-1 antibody or the anti-PD-L1 antibody.

[0388] The anti-CTLA4 antibody, and, when presents, the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered simultaneously.

[0389] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m², on day 1 of a first cycle of treatment, as a loading dose, and the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, and the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, on a subsequent day of a first cycle of treatment, as a loading dose.

[0390] According to an embodiment, the antibody-drug conjugate comprising the anti- CEACAM5 antibody may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m², on day 1 of at least on additional cycle(s) of treatment, as a subsequent dose, and the anti-CTLA4 antibody may be administered at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, and the anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, on a subsequent day of at least on additional cycle(s) of treatment, as a subsequent dose.

[0391 ] In some embodiments the loading dose and the subsequent dose are the same.

[0392] In some embodiments, the ADC may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m² on day 1 of a first cycle of treatment and of at least one subsequent cycle of treatment, and the anti-CTLA4 antibody and the anti- PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered, for the anti-CTLA4 antibody at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, and for the anti-PD-1 antibody or the anti-PD-L1 antibody at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, on a subsequent day of a first cycle of treatment and of at least one subsequent cycle of treatment.

[0393] In some embodiments, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered, for the anti-CTLA4 antibody at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, and for the anti-PD-1 antibody or the anti-PD-L1 antibody at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, on at least one subsequent day, for example on 1 to 20, or 2 to 18, or 4 to 16, or 6 to 14 or 8 to 12 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by one or a plurality of days of rest. [0394] In some embodiments, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered, for the anti-CTLA4 antibody at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, and for the anti-PD-1 antibody or the anti-PD-L1 antibody at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, on at least one subsequent day, for example on at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by one or a plurality of days of rest.

[0395] In some embodiments, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered, for the anti-CTLA4 antibody at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, and for the anti-PD-1 antibody or the anti-PD-L1 antibody at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, on at least one subsequent day, for example on at least 1 , 2, 3, 4, 5, or 6 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by 1 , 2, 3, 4, 6, or 7 of days of rest.

[0396] In some embodiments, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered, for the anti-CTLA4 antibody at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, and for the anti-PD-1 antibody or the anti-PD-L1 antibody at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, on at least one subsequent day, for example on at least 4 subsequent days of a first cycle and of at least one subsequent cycle of treatment. The plurality of subsequent days may be continuous or interrupted by 1 , 2, 3, 4, 6, or 7 of days of rest.

[0397] In some embodiments, a cycle may last three weeks, and the ADC may be administered at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m² on day 1 of a first cycle of treatment and of at least one subsequent cycle of treatment, and the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody, or the combination of the anti-CTLA4 antibody and of the anti-PD-1 antibody or the anti-PD-L1 antibody, may be administered, for the anti-CTLA4 antibody at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, and for the anti- PD-1 antibody or the anti-PD-L1 antibody at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, on day 10, 13, 17 and 20 of a first cycle of treatment and of at least one subsequent cycle of treatment.

[0398] In some embodiments, the anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody may be simultaneously administered as a combination.

[0399] According to some embodiments, in a use or method for treating a cancer, one cycle (first and additional cycles) may comprise the steps of:

[0400] i) administering the antibody-drug conjugate comprising an anti-CEACAM5- antibody at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m², once in the cycle, for example at day one of the cycle; and

[0401] ii) administering an anti-CTLA4 antibody at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, once in the cycle, for example at day one of the cycle.

[0402] The administration may be carried out on day one of a first and of at least one additional cycles of treatment.

[0403] The cycle is about two or three weeks.

[0404] The antibody-drug conjugate may be administered before the anti-CTLA4 antibody.

[0405] According to some embodiments, in a use or method for treating a cancer, one cycle (first and additional cycles) may comprise the steps of:

[0406] i) administering the antibody-drug conjugate comprising an anti-CEACAM5- antibody at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m², on day 1 of a first and of at least one additional cycles of treatment; and

[0407] ii) administering an anti-CTLA4 antibody at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, on at least one subsequent day of a first and of at least one additional cycles of treatment; [0408] The cycle is about two or three weeks.

[0409] According to some embodiments, in a use or method for treating a cancer, one cycle (first and additional cycles) may comprise the steps of:

[0410] i) administering the antibody-drug conjugate comprising an anti-CEACAM5- antibody at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m², once in the cycle, for example at day one of the cycle;

[0411] ii) administering an anti-CTLA4 antibody at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, once in the cycle, for example at day one of the cycle;

[0412] ill) administering an anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, once in the cycle, for example at day one of the cycle.

[0413] The administration may be carried out on day one of a first and of at least one additional cycles of treatment.

[0414] The cycle is about two or three weeks.

[0415] The antibody-drug conjugate may be administered before the anti-CTLA4 antibody.

[0416] The antibody-drug conjugate may be administered before the anti-PD-1 antibody or the anti-PD-L1 antibody.

[0417] The anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody may be simultaneously administered.

[0418] According to some embodiments, in a use or method for treating a cancer, one cycle (first and additional cycles) may comprise the steps of:

[0419] i) administering the antibody-drug conjugate comprising an anti-CEACAM5- antibody at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m², on day 1 of a first and of at least one additional cycles of treatment;

[0420] ii) administering an anti-CTLA4 antibody at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of about 10 mg/kg, on at least one subsequent day of a first and of at least one additional cycles of treatment; [0421] iii) administering an anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, on at least one subsequent day of a first and of at least one additional cycles of treatment.

[0422] The cycle is about two or three weeks.

[0423] The anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody may be simultaneously administered.

[0424] According to some embodiments, in a use or method for treating a cancer, one cycle (first and additional cycles) may comprise the steps of:

[0425] i) administering the antibody-drug conjugate comprising an anti-CEACAM5- antibody at a dose of about 60, 80, 100, 120, 135, 150, 170, 180, 190 or about 210 mg/m², on day 1 of a first and of at least one additional cycles of treatment;

[0426] ii) administering an anti-CTLA4 antibody at a dose of about 1 mg/kg, or at a dose of about 2.5 mg/kg, or at a dose of about 3 mg/kg, or at a dose of about 4 mg/kg, or at a dose of 6 mg/kg, or at a dose of 8 mg/kg, or at a dose of about 10 mg/kg, on day 10, 13, 17 and 20 of a first and of at least one additional cycles of treatment;

[0427] iii) administering an anti-PD-1 antibody or the anti-PD-L1 antibody may be administered at a dose of about 150 mg to about 400 mg, or at a dose of about 150 mg to about 300 mg or at a dose of about 200 mg, on day 10, 13, 17 and 20 of a first and of at least one additional cycles of treatment.

[0428] The cycle is about three weeks.

[0429] The anti-CTLA4 antibody and the anti-PD-1 antibody or the anti-PD-L1 antibody may be simultaneously administered.

Cancer

[0430] In an embodiment, the cancer is a carcinoma, a sarcoma or a blastoma. In a further embodiment, the cancer is a carcinoma.

[0431] According to an embodiment, the cancer is a cancer expressing CEACAM5. A cancer expressing CEACAM5 may also be named CEACAM5 positive cancer.

[0432] In some embodiments, the cancer is a CEACAM5-positive cancer. [0433] A CEACAM5-positive cancer is defined as cancer for which a CEACAM5 immunohistochemical [IHC] intensity is > 2+ in > 50% of cancer cells or > 2+ intensity in > 1% and < 50% of the cells tumors (or cancer cells).

[0434] In certain embodiments, the cancer has a negative or low CEACAM5 expression on tumor cells. A negative or low CEACAM5 expression on tumor cells defined as being a CEACAM5 immunohistochemical [IHC] intensity > 2+ in < 1 % of cells, as measured by immunohistochemistry (IHC).

[0435] In certain embodiments, the cancer has a moderate CEACAM5 expression on tumor cells. A moderate CEACAM5 expression on tumor cells may be defined as being a CEACAM5 immunohistochemical [IHC] intensity > 2+ in > 1% and in < 50% of cancer cells, as measured by immunohistochemistry.

[0436] In certain embodiments, the cancer has a high CEACAM5 expression on tumor cells. A high CEACAM5 expression on tumor cells may be defined as being a CEACAM5 immunohistochemical [IHC] intensity > 2+ intensity in > 50% of cancer cells, as measured by immunohistochemistry.

[0437] Immunohistochemical techniques for detecting antigens on cells or in tissue sections by means of immunological and chemical reactions are well-known in the field. Those techniques are highly sensitive and specific and can detect a wide variety of antigens. Immunohistochemistry methods comprise the following steps: binding of an antibody to a specific antigen; formation of an antibody-antigen complex by incubation with a secondary, enzyme-conjugated antibody, and generation of colored deposits at the sites of antibodyantigen binding in presence of substrate and chromogen catalyzed by the enzyme.

[0438] The CEACAM5 tumor expression may be determined by using Immunohistochemistry (IHC) assay. An assay may be done using anti-CEACAM5 antibody such as SANOFI’s antibody clone 769. Anti-CEACAM5 clone 769 is a murine monoclonal antibody with the same specificity as tusamitamab ravtansine to the CEACAM5 target. The assay may be run on Techmate platformer or on a Dako/Agilent Autostainer Link 48 IHC. Interpretation of CEACAM5 reactivity is to be performed using semi-quantitative Percent Scores (calculated by summing the percentages of intensities >2+) or H-score for CEACAM5 plasma membrane staining (whole or polarized) in tumor cells.

[0439] According to an embodiment, the cancer is selected from hepatocellular carcinoma, colorectal cancer, gastric cancer, gastroesophageal junction adenocarcinoma (GEJ), esophageal cancer, lung cancer (e.g., non-squamous non-small cell lung cancer), uterus cervix cancer, pancreatic cancer, ovarian cancer, thyroid cancer, bladder cancer, endometrial cancer, breast cancer, liver cancer, biliary tract cancer (for instance cholangiocarcinoma), prostate cancer, neuroendocrine cancer and skin cancer.

[0440] The cancer may be selected from a colorectal cancer, a gastric cancer, a gastroesophageal junction adenocarcinoma (GEJ), an esophageal cancer, a pancreatic cancer and a lung cancer.

[0441 ] In some embodiments, the cancer may be a colorectal cancer.

[0442] In some embodiments, the cancer may be a pancreatic cancer.

[0443] In some embodiments, the cancer may be selected from gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, esophageal cancer, and lung cancer.

[0444] In some embodiments, the cancer may be a gastric cancer, a gastroesophageal junction (GEJ) adenocarcinoma, or an esophageal cancer.

[0445] According to an embodiment, the cancer is gastric cancer or gastroesophageal junction adenocarcinoma (GEJ).

[0446] According to an embodiment, the cancer is a gastric cancer.

[0447] According to an embodiment, the cancer is a lung cancer.

[0448] A lung cancer may be a non-squamous non-small-cell lung cancer (NSQ NSCLC).

[0449] Non-small cell lung cancer is a disease in which malignant (cancer) cells form in the tissues of the lung. Smoking is the major cause of the disease. This is a type of epithelial lung cancer other than small cell lung carcinoma. There are several types of non-small cell lung cancer. Each type of non-small cell lung cancer has different kinds of cancer cells. The cancer cells of each type grow and spread in different ways. The types of non-small cell lung cancer are named for the kinds of cells found in the cancer and how the cells look under a microscope: (1 ) squamous cell carcinoma: Cancer that begins in squamous cells, which are thin, flat cells that look like fish scales. This is also called epidermoid carcinoma. (2) large cell carcinoma: Cancer that may begin in several types of large cells. (3) adenocarcinoma: Cancer that begins in the cells that line the alveoli and make substances such as mucus.

[0450] In some embodiments, the non-squamous non-small cell lung cancer may be an advanced or metastatic NSQ NSCLC.

[0451 ] According to an embodiment, the subject is a patient with malignant tumor, in particular with a malignant solid tumor, and more specifically with locally advanced or metastatic solid malignant tumor. A metastatic solid malignant tumor may be a metastatic cancer, for example a metastatic carcinoma. A cancer or a carcinoma may be as above indicated.

[0452] In some embodiments, the non-squamous non-small cell lung cancer has no epidermal growth factor receptor (EGFR) sensitizing mutation or v-raf murine sarcoma viral oncogene homolog B1 (BRAF) mutation or anaplastic lymphoma kinase/c-ros oncogene 1 (ALK/ROS) alterations.

Pharmaceutical compositions or combinations

[0453] The present disclosure also relates to a combination for the manufacture of a medicament for the treatment of cancer, the combination comprising an antibody-drug conjugate comprising an anti-CEACAM5-antibody and an anti-CTLA4 antibody.

[0454] The present disclosure also relates to a combination for the manufacture of a medicament for the treatment of cancer, the combination comprising an antibody-drug conjugate comprising an anti-CEACAM5-antibody, an anti-CTLA4 antibody, and an anti-PD-1 antibody or an anti-PD-L1 antibody.

[0455] The medicament may comprise the antibodies formulated individually, in separate containers.

[0456] In an embodiment, the present disclosure relates to a use of a combination of (i) an antibody-drug conjugate as disclosed herein and (ii) an anti-CTLA4 antibody, for the manufacture of a pharmaceutical composition or a kit, for the treatment of a cancer.

[0457] In an embodiment, the present disclosure relates to a use of a combination of (i) an antibody-drug conjugate as disclosed herein, (ii) an anti-CTLA4 antibody, and (iii) an anti-PD-1 antibody or anti-PD-L1 antibody, for the manufacture of a pharmaceutical composition or a kit, for the treatment of a cancer.

[0458] The pharmaceutical composition or the kit may comprise the antibodies formulated individually, in separate containers

[0459] In an embodiment, the combinations of the disclosure permit a simultaneous, separate or sequential administration of the antibody-drug conjugate comprising an anti- CEACAM5-antibody, the anti-CTLA4 antibody, and, when presents, the anti-PD-1 antibody or the anti-PD-L1 antibody.

[0460] In an embodiment, the combinations permit a sequential administration of the antibody-drug conjugate comprising an anti-CEACAM5-antibody, the anti-CTLA4 antibody, and, when presents, the anti-PD-1 antibody or the anti-PD-L1 antibody. [0461] In a further embodiment, combinations according to the disclosure are pharmaceutical compositions.

[0462] In a further embodiment, combinations according to the disclosure are pharmaceutical kits-of-parts.

[0463] In some embodiments, the present disclosure relates to a pharmaceutical composition comprising (i) an antibody-drug conjugate as disclosed herein, (ii) an anti-CTLA4 antibody, and a pharmaceutically acceptable excipient.

[0464] In some embodiments, (i) the antibody-drug conjugate, (ii) the anti-CTLA4 antibody, and (iii) the anti-PD-1 antibody or anti-PD-L1 antibody may be formulated in the form of two separate pharmaceutical compositions.

[0465] In some embodiments, the present disclosure relates to a pharmaceutical composition comprising (i) an antibody-drug conjugate as disclosed herein, (ii) an anti-CTLA4 antibody, (iii) an anti-PD-1 antibody or an anti-PD-L1 antibody, and a pharmaceutically acceptable excipient.

[0466] In some embodiments, (i) the antibody-drug conjugate, (ii) the anti-CTLA4 antibody, and, when presents, (iii) the anti-PD-1 antibody or anti-PD-L1 antibody may be formulated in a single pharmaceutical composition.

[0467] In some embodiments, (i) the antibody-drug conjugate, (ii) the anti-CTLA4 antibody, and (iii) the anti-PD-1 antibody or anti-PD-L1 antibody may be formulated in the form of two separate pharmaceutical compositions, where:

[0468] (a) one pharmaceutical composition may comprise the antibody-drug conjugate and the anti-CTLA4 antibody and the other pharmaceutical composition may comprise the anti- PD-1 antibody or anti-PD-L1 antibody, or

[0469] (b) one pharmaceutical composition may comprise the antibody-drug conjugate and the other pharmaceutical composition may comprise and the anti-CTLA4 antibody the anti- PD-1 antibody or anti-PD-L1 antibody, or

[0470] (c) one pharmaceutical composition may comprise the antibody-drug conjugate and the anti-PD-1 antibody or anti-PD-L1 antibody and the other pharmaceutical composition may comprise the anti-CTLA4 antibody.

[0471] In some embodiments, (i) the antibody-drug conjugate, (ii) the anti-CTLA4 antibody, and (iii) the anti-PD-1 antibody or anti-PD-L1 antibody may be formulated in the form of three separate pharmaceutical compositions, where (a) a first pharmaceutical composition may comprise the antibody-drug conjugate, (b) a second pharmaceutical composition may comprise the anti-CTLA4 antibody, and (c) a third pharmaceutical composition may comprise the anti-PD-1 antibody or anti-PD-L1 antibody.

[0472] In some embodiments, the present disclosure relates to a kit comprising (i) a pharmaceutical composition comprising an antibody-drug conjugate as disclosed herein and a pharmaceutically acceptable excipient, and (ii) a pharmaceutical composition comprising an anti-CTLA4 antibody and a pharmaceutically acceptable excipient.

[0473] In some embodiments, the present disclosure relates to a kit comprising (i) a pharmaceutical composition comprising an antibody-drug conjugate as disclosed herein and a pharmaceutically acceptable excipient, (ii) a pharmaceutical composition comprising an anti- CTLA4 antibody and a pharmaceutically acceptable excipient, and (iii) a pharmaceutical composition comprising an anti-PD-1 antibody or an anti-PD-L1 antibody and a pharmaceutically acceptable excipient.

[0474] In some embodiments, the present disclosure relates to a pharmaceutical composition as disclosed herein, or a kit as disclosed herein, for a use for treating cancer.

[0475] In some embodiments, in the uses as disclosed herein, the administration of the antibody-drug conjugate comprising an anti-CEACAM5-antibody and/or the anti-CTLA4 antibody and/or the anti-PD-1 antibody or the anti-PD-L1 antibody may be carried out by parenteral route. A suitable parenteral route may be intravenous infusion.

[0476] "Pharmaceutical excipient" or "pharmaceutically acceptable excipient" refers to molecular entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to a mammal, especially a human, as appropriate. A pharmaceutically acceptable carrier or excipient refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type.

[0477] As used herein, “pharmaceutically-acceptable carriers or excipients” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, and the like that are physiologically compatible. Examples of suitable carriers, diluents and/or excipients include one or more of water, amino acids, saline, phosphate buffered saline, buffer phosphate, acetate, citrate, succinate; amino acids and derivates such as histidine, arginine, glycine, proline, glycylglycine; inorganic salts NaCI, calcium chloride; sugars or polyalcohols such as dextrose, glycerol, ethanol, sucrose, trehalose, mannitol; surfactants such as Polysorbate 80, polysorbate 20, poloxamer 188; and the like, as well as combination thereof. In many cases, it will be preferable to include isotonic agents, such as sugars, polyalcohols, or sodium chloride in the composition, and formulation may also contain an antioxidant such as tryptamine and a stabilizing agent such as Tween 20.

[0478] The form of the pharmaceutical compositions, the route of administration, the dosage and the regimen naturally depend upon the condition to be treated, the severity of the illness, the age, weight, and gender of the subject, etc.

[0479] The pharmaceutical compositions of the disclosure can be formulated for a topical, oral, parenteral, intranasal, intravenous, intramuscular, subcutaneous or intraocular administration and the like. In an embodiment, the pharmaceutical compositions and combinations of the disclosure are formulated for intravenous administration.

[0480] In particular, the pharmaceutical compositions contain vehicles or excipients, which are pharmaceutically acceptable for a formulation capable of being injected. These may be in particular isotonic, sterile, saline solutions (monosodium or disodium phosphate, sodium, potassium, calcium or magnesium chloride and the like or mixtures of such salts), or dry, especially freeze-dried compositions which upon addition, depending on the case, of sterilized water or physiological saline, permit the constitution of injectable solutions.

[0481] The pharmaceutical composition can be administrated through drug combination devices.

[0482] The doses used for the administration can be adapted as a function of various parameters, and in particular as a function of the mode of used, of the relevant pathology, or alternatively of the desired duration of treatment.

[0483] To prepare pharmaceutical compositions, an effective amount of antibody-drug conjugate comprising an anti-CEACAM5-antibody and/or of an anti-CTLA4 antibody and/or of an anti-PD-1 antibody or anti-PD-L1 antibody may be dissolved or dispersed in a pharmaceutically acceptable carrier or aqueous medium.

[0484] The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; formulations including sesame oil, peanut oil or aqueous propylene glycol; and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and injectable with the appropriate device or system for delivery without degradation. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi.

[0485] Solutions of the active compounds as free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

[0486] The antibody-drug conjugate comprising an anti-CEACAM5-antibody can be formulated into a composition in a neutral or salt form. Pharmaceutically acceptable salts include the acid addition salts (formed with the free amino groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, glycine, histidine, procaine and the like.

[0487] The carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetables oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the subsequent of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminium monostearate and gelatin.

[0488] Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum-drying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. [0489] The preparation of more, or highly concentrated solutions for direct injection is also contemplated, where the use of DMSO as solvent is envisioned to result in extremely rapid penetration, delivering high concentrations of the active agents to a small tumor area.

[0490] Upon formulation, solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective. The formulations are easily administered in a variety of dosage forms, such as the type of injectable solutions described above, but drug release capsules and the like can also be employed.

[0491] For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. In this connection, sterile aqueous media which can be employed will be known to those of skill in the art in light of the present disclosure. For example, one dosage could be dissolved in 1 ml of isotonic NaCI solution and either added to 1000 ml of hypodermoclysis fluid or injected at the proposed site of infusion, (see for example, "Remington's Pharmaceutical Sciences" 15th Edition, pages 1035-1038 and 1570-1580). Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject.

[0492] The antibody-drug conjugate comprising an anti-CEACAM5-antibody formulated for parenteral administration, such as intravenous or intramuscular injection, other pharmaceutically acceptable forms include, e.g., tablets or other solids for oral administration; time release capsules; and any other form currently used.

[0493] In certain embodiments, the use of liposomes and/or nanoparticles is contemplated for the introduction of polypeptides into host cells. The formation and use of liposomes and/or nanoparticles are known to those of skill in the art.

[0494] Nanocapsules can generally entrap compounds in a stable and reproducible way. To avoid side effects due to intracellular polymeric overloading, such ultrafine particles (sized around 0.1 pm) are generally designed using polymers able to be degraded in vivo. Biodegradable polyalkyl-cyanoacrylate nanoparticles, or biodegradable polylactide or polylactide co glycolide nanoparticules that meet these requirements are contemplated for use in the present disclosure, and such particles may be easily made.

[0495] Liposomes are formed from phospholipids that are dispersed in an aqueous medium and spontaneously form multilamellar concentric bilayer vesicles (also termed multilamellar vesicles (MLVs)). MLVs generally have diameters of from 25 nm to 4 pm. Sonication of MLVs results in the formation of small unilamellar vesicles (SLIVs) with diameters in the range of 200 to 500 A, containing an aqueous solution in the core. The physical characteristics of liposomes depend on pH, ionic strength and the presence of divalent cations.

[REFERENCES]

Schofield et al, Activity of murine surrogate antibodies for durvalumab and tremelimumab lacking effector function and the ability to deplete regulatory T cells in mouse models of cancer, mAbs, 2021 ;13:1 Magiera-Mularz et al, Human and mouse PD-L1 : similar molecular structure, but different druggability profiles (iScience 2021 ; 24)

Plowman J, Dykes DJ, Hollingshead M, Simpson-Herren L and Alley MC. Human tumor xenograft models in NCI drug development. In: Feibig HH BA, editor. Basel: Karger.; 1999 p 101 -125

[EXAMPLES]

[0496] The following examples illustrate the embodiments of the invention that are presently best known. However, it is to be understood that the following are only exemplary or illustrative of the application of the principles of the present invention. Numerous modifications and alternative compositions, methods, and systems may be devised by those skilled in the art without departing from the spirit and scope of the present invention. Thus, while the present invention has been described above with particularity, the following examples provide further detail in connection with what are presently deemed to be the most practical and preferred embodiments of the invention.

Preliminary remarks

[0497] Preclinical studies of PD-1 , PD-L1 and/or CTLA-4 blockade have relied heavily on mouse syngeneic tumor models with intact immune systems, which facilitate dissection of immunosuppressive mechanisms in the tumor microenvironment. Commercially developed monoclonal antibodies (mAbs) targeting human PD-L1 and PD-1 may not demonstrate cross- reactive binding to their mouse orthologs, and surrogate anti-mouse antibodies are often used in their place to inhibit these immune checkpoints (Schofield et al, Activity of murine surrogate antibodies for durvalumab and tremelimumab lacking effector function and the ability to deplete regulatory T cells in mouse models of cancer, mAbs, 2021 ;13:1 ). They are functionally equivalent to the therapeutic human antibodies, but they are not strictly equivalent (IgG isotype, level of affinity, biological activity, effector function, ...). For these preclinical investigations, the anti muPD-1 surrogate mAb, clone RMP1 -14 (rat lgG1 anti-muPD-1 ), the anti muCTLA-4 surrogate mAb, clone 9D9, and the anti-huPD-L1 mAb, Atezolizumab, that is able to bind and blocks mouse PD-L1 (Magiera-Mularz et al, Human and mouse PD-L1 : similar molecular structure, but different druggability profiles (iScience 2021 ; 24) were used.

[0498] In addition, CEACAM5 protein is not expressed in rodents and human CEACAM5 engineered murine tumors do not grow in immunocompetent mice, reason why these experiments are conducted with the huMAb2-3-SPDB-DM4 ADC, which, at dose high enough, to deliver the payload in a non-specific manner. It will be administered at high dose to exploit the enhanced permeability and retention effect observed for solid tumors subcutaneously implanted in mice that leads to selective delivery of macromolecular drugs to the tumor site. Dose have been modulated to obtain different levels of antitumoral activity (inactivity, moderate activity and/or high activity mimicking the different response levels expected in tumor with low, moderate or high CEACAM5 expression, reciprocally).

[0499] The first used syngeneic tumor model, MC38 was isolated from a colon tumor in a C57BL/6 mouse following long term exposure to the carcinogen DMH (1 ,2- dimethylhydrazine dihydrochloride). MC38 was an hypermutated and microsatellite-instable colorectal cancer and showed a favorable response profile to immunomodulatory antibodies suggesting a tumor microenvironment amendable to immune activation. In this immune checkpoint inhibitors (ICI) sensitive model, sub-optimal dose of each blocking mAb (5 mg/kg) were used for the IO co-treatment.

[0500] The second one, CT26 was isolated from a colon tumor in a BALB/c mouse following long term exposure to the carcinogen N-nitroso-N-methylurea. CT26 was a nonhypermutated colorectal cancer and was only partially sensitive immunomodulatory antibodies. In this ICI poorly sensitive model, optimal dose of each blocking mAb (10 mg//kg) were used for the ICI co-treatment.

Example 1 : Activity of the immunoconjugate huMAb2-3-SPDB-DM4 and the anti- muPD-1 + anti-muCTLA-4 antibody co-treatment, as single agents or in combination against subcutaneous colon MC38 syngeneic tumor model in C57BI/6 mice

Experimental procedure

[0501] The activity of huMAb2-3-SPDB-DM4 and the anti-muPD-1/anti-muCTLA-4 antibodies, was evaluated as single agent or in combination in a subcutaneous colon MC38 syngeneic tumor, implanted s.c. in female C57BI/6 mice. Control groups were left untreated. The doses of the compounds used are given in mg/kg.

[0502] Mice were randomized in 6 groups (n = 6) when tumors were well established (median tumour burden reached a median tumor volume of 157 mm³). huMAb2-3-SPDB-DM4 was administered at 15 and 25 mg/kg following a single IV administration on day 10 and the anti-muPD-1 and anti-muCTLA-4 antibodies were co-administered at 5 mg/kg following IV administrations on days 10, 13, 17 and 20.

[0503] For the evaluation of anti-tumor activity, animals were weighed, and tumors were measured by caliper 2 times weekly. A dosage producing a 20% weight loss at nadir (mean of group) or 10% or more drug deaths, was considered an excessively toxic dosage. Animal body weights included the tumor weights. Tumor volumes were calculated using the formula mass (mm³) = [length (mm) x width (mm) x width (mm)]/2. The primary efficacy end points are AT/AC, percent median regression, partial and complete regressions (PR and CR).

[0504] Changes in tumor volume for each treated (T) and control (C) are calculated for each tumor by subtracting the tumor volume on the day of first treatment (staging day) from the tumor volume on the specified observation day. The median AT is calculated for the treated group and the median AC is calculated for the control group. Then the ratio AT/AC is calculated and expressed as a percentage: AT/AC = (delta T/delta C) x 100.

[0505] The dose is considered as therapeutically active when AT/ AC is lower than 40% and very active when AT/ AC is lower than 10%. If AT/ AC is lower than 0, the dose is considered as highly active, and the percentage of regression is dated (Plowman J, Dykes DJ, Hollingshead M, Simpson-Herren L and Alley MC. Human tumor xenograft models in NCI drug development. In: Feibig HH BA, editor. Basel: Karger.; 1999 p 101 -125):

[0506] % tumor regression is defined as the % of tumor volume decrease in the treated group at a specified observation day compared to its volume on the first day of first treatment.

[0507] At a specific time point and for each animal, % regression is calculated. The median % regression is then calculated for the group: volume_t0 -volume,_0Q

[0508] % regression (at t) = volume_t0

[0509] Partial regression (PR): Regressions are defined as partial if the tumor volume decreases to 50 % of the tumor volume at the start of treatment.

[0510] Complete regression (CR): Complete regression is achieved when tumor volume = 0 mm³ (CR is considered when tumor volume cannot be recorded).

Results

[0511] The results for efficacy evaluation of the immunoconjugate huMAb2-3-SPDB- DM4 and the anti-muPD-1/anti-muCTLA-4 antibodies, as single agents or in combination against subcutaneous colon MC38 syngeneic tumor model in C57BI/6 mice are presented on Figure 1 and Table 1.

[0512] HuMAb2-3-SPDB-DM4 and anti-muPD-1/anti-muCTLA-4 mAbs were administered at doses lower than maximal tolerated dose (MTD) and treatments were well tolerated and did not induce toxicity. [0513] The huMAb2-3-SPDB-DM4 as a single agent was very active at 25 mg/kg with a AT/AC at D24 equal to 10% (no significant vs control), 2/6 PR and 1/6 CR and inactive at 15 mg/kg with a AT/AC at D24 equal to 48% (no significant vs control) and 1/6 PR.

[0514] The anti-muPD-1/anti-muCTLA-4 mAbs as single agent were active with a AT/AC at D24 equal to 24% (no significant vs control).

[0515] The combination between huMAb2-3-SPDB-DM4 at 25 mg/kg and anti-muPD- 1/anti-muCTLA-4 mAbs was highly active with a AT/AC at D24 inferior to 0% (p = 0.0034 vs control), a tumor regression of 50%, 4/6 PR and 3/6 CR.

[0516] The combination between huMAb2-3-SPDB-DM4 at 15 mg/kg and anti-muPD- 1 /anti-muCTLA-4 mAbs was active with a AT/AC at D24 equal to 10% (p = 0.0084 vs control), a tumor regression of 100%, 4/6 PR and 4/6 CR.

[0517] In conclusion to the experiment in the colon MC38 syngeneic tumor, huMAb2- 3-SPDB-DM4, when it was active or inactive, synergized with anti-muPD-1 /anti-muCTLA-4 mAbs, in sub-optimal conditions, leading to a robust activity and to tumor regression.

Table 1 : Activity of huMAb2-3-SPDB-DM4 and anti-muPD-1 /anti-CTLA-4, as single agents or in combination in a subcutaneous colon MC38 syngeneic tumor, implanted s.c. in female C57BI/6 mice

Agent Route Dosage in Schedule Drug death Mean body mg/kg in day (day) weight change

(total) in % at nadir

(day) .. n n 25 (25) 10 0/6 +1.45 (D13) huMAb2-3-

IV

SPDB-DM4

15 (15) 10 0/6 +1.22 (D13) anti-muPD-1 / 5 (20) 10, 13, 17,

IV 0/6 +1.12 (D13) anti-CTLA-4 5 (20) 20

10 huMAb2-3- IV 25 (25)

10, 13, 17, 0/6 +2.45 (D13)

SPDB-DM4 IV 5 (20)

20 Agent Route Dosage in Schedule Drug death Mean body mg/kg in day (day) weight change

(total) in % at nadir

(day)

+ anti-muPD- 10

15 (15)

1/anti-CTLA-4 10, 13, 17, 0/6 +0.63 (D13)

5 (20)

20

Control - - - - +1.22 (D13)

Table 1 continued

Agent Median Median % Regression Biosatitic Biological

AT/AC in of PR CR P value³ comments

% (D24) regression (D24)

(day)

, .... 10 - 2/6 1/6 ns Very active huMAb2-3-

SPDB-DM4 48 - 1/6 0/6 ns Inactive anti-muPD-1 /

24 - 2/6 1/6 ns active anti-CTLA-4 huMAb2-3- < 0 50 (D27) 4/6 3/6 = 0.0034 Highly active

SPDB-DM4

+ anti-muPD- 10 100 (D35) 4/6 4/6 = 0.0084 Very active

1 /anti-CTLA-4

Control _ _ _ _

[0518]^a: Statistical analysis. The p-values were obtained using a contrast analysis to compare each treated group versus control using Bonferroni-Holm adjustment for multiplicity after a two-way Anova-Type with repeated measures on tumor volume changes from baseline. A probability less than 5% (p<0.05) was considered as significant.

[0519] AT/AC = ratio of medians of tumor volume changes from baseline between treated and control groups; PR = Partial regression; CR = Complete regression. Example 2: Activity of the immunoconjugate huMAb2-3-SPDB-DM4 and the anti- mu/huPD-L1/anti-muCTLA-4 antibody co-treatment, as single agents or in combination against subcutaneous colon MC38 syngeneic tumor model in C57BI/6 mice.

Experimental procedure

[0520] The activity of huMAb2-3-SPDB-DM4 and the anti-mu/huPD-L1/anti-muCTLA- 4 antibodies, was evaluated as single agent or in combination in a subcutaneous colon MC38 syngeneic tumor, implanted s.c. in female C57BI/6 mice. Control groups were left untreated. The doses of the compounds used are given in mg/kg.

[0521] Mice were randomized in 6 groups (n = 6) on day 10 post tumour implantation when median tumour burden reached a median tumor volume of 157 mm³. huMAb2-3-SPDB- DM4 was administered at 15 and 25 mg/kg following a single IV administration on day 10 and the anti-mu/huPD-L1/anti-muCTLA-4 mAbs were co-administered at 5 mg/kg following IV administrations on days 10, 13, 17 and 20.

[0522] See above, Example 1 , for the conditions of anti-tumor activity and toxicity evaluation.

Results

[0523] The results for efficacy evaluation of the immunoconjugate huMAb2-3-SPDB- DM4 and the anti-mu/huPD-L1 antibody, as single agents or in combination against subcutaneous colon MC38 syngeneic tumor model in C57BI/6 mice are presented on Figure 2 and Table 2.

[0524] HuMAb2-3-SPDB-DM4 and anti-mu/huPD-L1/anti-muCTLA-4 mAbs were administered at doses lower than maximal tolerated dose (MTD) and treatments were well tolerated and did not induce toxicity.

[0525] The huMAb2-3-SPDB-DM4 as a single agent was highly active at 25 mg/kg with a AT/AC at D24 equal to 10% (no significant vs control), 2/6 PR and 1/6 CR and inactive at 15 mg/kg with a AT/AC at D24 equal to 48% (no significant vs control) and 1/6 PR.

[0526] The anti-mu/huPD-L1/anti-muCTLA-4 mAbs as single agent were active with a AT/AC at D24 equal to 12% (no significant vs control), 3/6 PR and 3/6 CR.

[0527] The combination between huMAb2-3-SPDB-DM4 at 25 mg/kg and anti- mu/huPD-L1/anti-muCTLA-4 mAbs was highly active with a AT/AC at D24 inferior to 0% (p <0.0001 vs control), a tumor regression of 100%, 6/6 PR and 5/6 CR. [0528] The combination between huMAb2-3-SPDB-DM4 at 15 mg/kg and anti- mu/huPD-L1/anti-muCTLA-4 mAbs was very active a AT/AC at D24 inferior to 0% (p <0.0001 vs control), a tumor regression of 89%, 5/6 PR and 3/6 CR.

[0529] In conclusion to the experiment in the colon MC38 syngeneic tumor, huMAb2- 3-SPDB-DM4, when it was active and inactive, synergized with anti-mu/huPD-L1/muCTLA-4 mAbs, in sub-optimal conditions, leading to robust activity and complete tumor regression.

Table 2: Activity of huMAb2-3-SPDB-DM4 and anti-mu/huPD-L1/anti-CTLA-4, as single agent or in combination in a subcutaneous colon MC38 syngeneic tumor, implanted s.c. in female C57BI/6 mice

Agent Route Dosage Schedule Drug Mean body weight in in day death change in % at nadir mg/kg (day) (day)

(total) .. A n n rn n 25 (25) 10 0/6 +1.45 (D13) huMAb2-3-SPDB-

IV

DM4

15 (15) 10 0/6 +1.22 (D13) anti- 5 (20) 10, 13, mu/huPD-L1/anti- IV 0/6 +0.78 (D13)

5 (20) 17, 20 muCTLA-4

10 25 (25) huMAb2-3-SPDB- 10, 13, 0/6 +0.03 (D13)

5 (20)

DM4 IV 17, 20

+ anti-mu/huPD- IV 10

15 (15) L1/anti-muCTLA-4 10, 13, 0/6 +2.90 (D13)

5 (20)

17, 20

Control - - - - +1.22 (D13)

Table 2 continued Agent Median Median % of Regression Biosatitic Biological

AT/AC regression PR CR P value³ comments in % (day) (D24)

(D24)

. ...._{n n} 10 - 2/6 1/6 ns Very active huMAb2-3-SPDB-

DM4

48 - 1/6 0/6 ns Inactive anti- mu/huPD-L1/anti- 12 - 3/6 3/6 ns Active muCTLA-4 huMAb2-3-SPDB- < 0 100 (D24) 6/6 5/6 < 0.0001 Highly active

DM4

+ anti-mu/huPD- < 0 89 (D41 ) 5/6 3/6 < 0.0001 Highly active

L1 /anti-muCTLA-4

Control _ _ _ _

[0530]^a: Statistical analysis. The p-values were obtained using a contrast analysis to compare each treated group versus control using Bonferroni-Holm adjustment for multiplicity after a two-way Anova-Type with repeated measures on tumor volume changes from baseline. A probability less than 5% (p<0.05) was considered as significant. [0531] AT/AC = ratio of medians of tumor volume changes from baseline between treated and control groups; PR = Partial regression; CR = Complete regression

Example 3: Activity of the immunoconjugate huMAb2-3-SPDB-DM4 and anti- mu CT LA-4 antibody, as single agents or in combination against subcutaneous colon CT26 syngeneic tumor model in BALB/c mice.

Experimental procedure

[0532] The activity of huMAb2-3-SPDB-DM4 and the anti-muCTLA-4 antibody, was evaluated as single agent or in combination in a subcutaneous colon CT26 syngeneic tumor, implanted s.c. in female BALB/c mice. Control groups were left untreated. The doses of the compounds used are given in mg/kg.

[0533] Mice were randomized in 6 groups (n = 8) when tumors were well established (median tumour burden reached a median tumor volume of 161 mm³). huMAb2-3-SPDB-DM4 was administered at 35 and 25 mg/kg following a single IV administration on day 10 and the anti-muCTLA-4 antibodies was administered at 10 mg/kg following IV administrations on days 10, 13 and 17 as single agent and at 13, 17 and 20 in combination.

[0534] See above (Example 1 ) for the conditions of anti-tumor activity and toxicity evaluation.

Results

[0535] The results for efficacy evaluation of the immunoconjugate huMAb2-3-SPDB- DM4 and the anti-muCTLA-4 antibody, as single agents or in combination against subcutaneous colon CT26 syngeneic tumor model in BALB/C mice are presented on Figure 3 and Table 3.

[0536] HuMAb2-3-SPDB-DM4 and anti-muCTLA-4 mAb were administered at doses lower than maximal tolerated dose (MTD) and treatments were well tolerated and did not induce toxicity.

[0537] The huMAb2-3-SPDB-DM4 as a single agent was active at 35 mg/kg with a AT/AC at D21 equal to 11% (p < 0.0001 vs control) and inactive at 25 mg/kg with a AT/AC at D21 equal to 41% (no significant vs control).

[0538] The anti-muCTLA-4 mAb as single agent were inactive with a AT/AC at D21 equal to 52% (no significant vs control).

[0539] The combination between huMAb2-3-SPDB-DM4 at 35 mg/kg and anti- muCTLA-4 mAb was very active with a AT/AC at D21 equal to 4% (p < 0.0001 vs control), 3/8 PR and 2/8 CR.

[0540] The combination between huMAb2-3-SPDB-DM4 at 25 mg/kg and anti- muCTLA-4 mAb was active with a AT/AC at D21 equal to 13% (p < 0.0001 vs control), 2/8 PR and 2/8 CR.

[0541] In conclusion to the experiment in the colon CT26 syngeneic tumor, huMAb2-3- SPDB-DM4, when it was active or inactive, synergized with anti-muCTLA-4 mAb leading to a robust activity and to some complete tumor regression.

Table 3: Activity of huMAb2-3-SPDB-DM4 and anti-CTLA-4, as single agents or in combination in a subcutaneous colon CT26 syngeneic tumor, implanted s.c. in female BALB/c mice Agent Route Dosage Schedule Drug Mean body weight in in day death change in % at nadir mg/kg (day) (day)

(total)

. .... 35 (35) 10 0/8 -0.63 (D13) huMAb2-3-

IV

SPDB-DM4

25 (25) 10 0/8 -0.33 (D14) anti-CTLA-4 IV 10 (30) 10. 13, 17 0/8 -0.80 (D14)

35 (35) 10 huMAb2-3- 0/8 -0.30 (D17)

IV 10 (30) 13, 17, 20

SPDB-DM4

IV 25 (25) 10

+ anti-CTLA-4 0/8 +2.19 (D14)

10 (30) 13, 17, 20

Control - - - - -0.01 (D14)

Table 3 continued

Agent Median Median % Regression Biosatitic Biological

AT/AC in % of PR CR P value³ comments

(D21) regression (D21)

(day)

. .... 11 - 0/8 0/8 < 0.0001 Active huMAb2-3-

SPDB-DM4

41 - 0/8 0/8 ns Inactive anti-CTLA-4 52 - 0/8 0/8 ns Inactive huMAb2-3- 4 - 3/8 2/8 < 0.0001 Very active

SPDB-DM4

13 - 2/8 2/8 < 0.0001 Active

+ anti-CTLA-4

Control _ _ _ _

[0542]^a: Statistical analysis. The p-values were obtained using a contrast analysis to compare each treated group versus control using Bonferroni-Holm adjustment for multiplicity after a two-way Anova-Type with repeated measures on tumor volume changes from baseline. A probability less than 5% (p<0.05) was considered as significant. [0543] AT/AC = ratio of medians of tumor volume changes from baseline between treated and control groups; PR = Partial regression; CR = Complete regression

Example 4: Activity of the immunoconjugate huMAb2-3-SPDB-DM4 and the anti- muPD-1 + anti-muCTLA-4 antibody co-treatment, as single agents or in combination against subcutaneous colon CT26 syngeneic tumor model in BALB/c mice.

Experimental procedure

[0544] The activity of huMAb2-3-SPDB-DM4 and the anti-muPD-1 /anti-muCTLA-4 antibodies, was evaluated as single agent or in combination in a subcutaneous colon CT26 syngeneic tumor, implanted s.c. in female BALB/c mice. Control groups were left untreated. The doses of the compounds used are given in mg/kg.

[0545] Mice were randomized in 6 groups (n = 8) when tumors were well established (median tumour burden reached a median tumor volume of 151 mm³). huMAb2-3-SPDB-DM4 was administered at 15 and 25 mg/kg following a single IV administration on day 10 and the anti-muPD-1 and anti-muCTLA-4 antibodies were co-administered at 10 mg/kg following IV administrations on days 10, 13, 17 and 20.

[0546] See above (Example 1 ) for the conditions of anti-tumor activity and toxicity evaluation.

Results

[0547] The results for efficacy evaluation of the immunoconjugate huMAb2-3-SPDB- DM4 and the anti-muPD-1 /anti-muCTLA-4 antibodies, as single agents or in combination against subcutaneous colon CT26 syngeneic tumor model in BALB/C mice are presented on Figure 4 and Table 4.

[0548] HuMAb2-3-SPDB-DM4 and anti-muPD-1 /anti-muCTLA-4 mAbs were administered at doses lower than maximal tolerated dose (MTD) and treatments were well tolerated and did not induce toxicity.

[0549] The huMAb2-3-SPDB-DM4 as a single agent was active at 25 mg/kg with a AT/AC at D20 equal to 20% (no significant vs control), 4/8 PR and 2/8 CR and inactive at 15 mg/kg with a AT/AC at D20 equal to 42% (no significant vs control).

[0550] The anti-muPD-1 /anti-muCTLA-4 mAbs as single agent were active with a AT/AC at D20 equal to 19% (no significant vs control), 2/8 PR and 2/8 CR. [0551] The combination between huMAb2-3-SPDB-DM4 at 25 mg/kg and anti-muPD- 1/anti-muCTLA-4 mAbs was highly active with a AT/AC at D20 (last day of control group presence) equal to 11% (p = 0.0002 vs control), a tumor regression of 100%, 8/8 PR and 7/8 CR. [0552] The combination between huMAb2-3-SPDB-DM4 at 15 mg/kg and anti-muPD-

1/anti-muCTLA-4 mAbs was highly active with a AT/AC at D20 (last day of control group presence) equal to 7% (p < 0.0001 vs control), a tumor regression of 100%, 6/8 PR and 6/8 CR.

[0553] In conclusion to the experiment in the colon CT26 syngeneic tumor, huMAb2-3- SPDB-DM4, when it was active or inactive, synergized with anti-muPD-1/anti-muCTLA-4 mAbs leading to a robust activity and to complete tumor regression.

Table 4: Activity of huMAb2-3-SPDB-DM4 and anti-CTLA-4, as single agents or in combination in a subcutaneous colon CT26 syngeneic tumor, implanted s.c. in female BALB/c mice

Agent Route Dosage Schedule Drug Mean body weight in in day death change in % at nadir mg/kg (day) (day)

(total) .. A n n rn n 25 (25) 10 0/8 -0.26 (D11) huMAb2-3-SPDB-

IV

DM4

15 (15) 10 0/8 +0.44 (D11) anti-muPD-1 / anti- 10 (40) 10, 13,

IV 0/8 +0.30 (D11)

CTLA-4 10 (40) 17, 20

10

25 (25) huMAb2-3-SPDB- 10, 13, 0/8 -1.21 (D11)

10 (40)

DM4 IV 17, 20

+ anti-muPD-1 /anti- IV 10

15 (15)

CTLA-4 10, 13, 0/8 -0.41 (D11)

10 (40)

17, 20

Control - - - - -0.03 (D11) Table 4 continued

Agent Median Median % of Regression Biosatitic Biological

AT/AC in regression PR CR P value³ comments % (D20) (day) (D20)

. .... 20 - 4/8 2/8 ns Active huMAb2-3-

SPDB-DM4

42 - 0/8 0/8 ns Inactive anti-muPD-1 /

19 - 2/8 2/8 ns Active anti-CTLA-4 huMAb2-3- 11 100 (D39) 8/8 7/8 = 0.0002 Highly active

SPDB-DM4

+ anti-muPD- 7 100 (D35) 6/8 6/8 < 0.0001 Highly active

1 /anti-CTLA-4

Control _ _ _ _

[0554]^a: Statistical analysis. The p-values were obtained using a contrast analysis to compare each treated group versus control using Bonferroni-Holm adjustment for multiplicity after a two-way Anova-Type with repeated measures on tumor volume changes from baseline. A probability less than 5% (p<0.05) was considered as significant.

[0555] AT/AC = ratio of medians of tumor volume changes from baseline between treated and control groups; PR = Partial regression; CR = Complete regression

Example 5: Activity of the immunoconjugate huMAb2-3-SPDB-DM4 and the anti- mu/huPD-L1/anti-muCTLA-4 antibody co-treatment, as single agents or in combination against subcutaneous colon CT26 syngeneic tumor model in BALB/c mice.

Experimental procedure

[0556] The activity of huMAb2-3-SPDB-DM4 and the anti-mu/huPD-L1/anti-muCTLA- 4 antibodies, was evaluated as single agent or in combination in a subcutaneous colon CT26 syngeneic tumor, implanted s.c. in female BALB/c mice. Control groups were left untreated. The doses of the compounds used are given in mg/kg.

[0557] Mice were randomized in 6 groups (n = 8) when tumors were well established (median tumour burden reached a median tumor volume of 151 mm³). huMAb2-3-SPDB-DM4 was administered at 15 and 25 mg/kg following a single IV administration on day 10 and the anti-mu/huPD-L1/anti-muCTLA-4 mAbs were co-administered at 10 mg/kg following IV administrations on days 10, 13, 17 and 20.

[0558] See above (Example 1) for the conditions of anti-tumor activity and toxicity evaluation.

Results

[0559] The results for efficacy evaluation of the immunoconjugate huMAb2-3-SPDB- DM4 and the anti-mu/huPD-L1 antibody, as single agents or in combination against subcutaneous colon CT26 syngeneic tumor model in BALB/c mice are presented on Figure 5 and Table 5.

[0560] HuMAb2-3-SPDB-DM4 and anti-mu/huPD-L1/anti-muCTLA-4 mAbs were administered at doses lower than maximal tolerated dose (MTD) and treatments were well tolerated and did not induce toxicity.

[0561] The huMAb2-3-SPDB-DM4 as a single agent was active at 25 mg/kg with a AT/AC at D20 equal to 20% (no significant vs control), 4/8 PR and 2/8 CR and inactive at 15 mg/kg with a AT/AC at D20 equal to 42% (no significant vs control).

[0562] The anti-muPD-1/anti-muCTLA-4 mAbs as single agent were active with a AT/AC at D20 equal to 19% (no significant vs control), 2/8 PR and 2/8 CR.

[0563] The combination between huMAb2-3-SPDB-DM4 at 25 mg/kg and anti- mu/huPD-L1/anti-muCTLA-4 mAbs was highly active with a AT/AC at D20 (last day of control group presence) equal to 20% (p = 0.0321 vs control), a tumor regression of 100%, 8/8 PR and 7/8 CR.

[0564] The combination between huMAb2-3-SPDB-DM4 at 15 mg/kg and anti- mu/huPD-L1/anti-muCTLA-4 mAbs was highly active with a AT/AC at D20 (last day of control group presence) equal to 14% (p = 0.0014 vs control), a tumor regression of 100%, 5/8 PR and 5/8 CR.

[0565] In conclusion to the experiment in the colon CT26 syngeneic tumor, huMAb2-3- SPDB-DM4, when it was active and inactive, synergized with anti-mu/huPD-L1/anti-muCTLA- 4 mAbs leading to robust activity and complete tumor regression. Table 5: Activity of huMAb2-3-SPDB-DM4 and anti-mu/huPD-L1/anti-CTLA-4, as single agent or in combination in a subcutaneous colon CT26 syngeneic tumor, implanted s.c. in female BALB/c mice

Agent Route Dosage Schedule Drug Mean body weight in mg/kg in day death change in % at nadir

(total) (day) (day) .. n n 25 (25) 10 0/8 -0.26 (D11) huMAb2-3-

IV

SPDB-DM4

15 (15) 10 0/8 +0.44 (D11) anti- mu/huPD- 10 (40) 10, 13,

IV 0/8 -0.31 (D11)

L1/anti- 10 (40) 17, 20 muCTLA-4

10 huMAb2-3- 25 (25)

10, 13, 0/8 -0.11 (D11)

SPDB-DM4 10 (40)

IV 17, 20

+ anti-mu/huPD-

IV 10

L1/anti- 15 (15)

10, 13, 0/8 +0.54 (D11) muCTLA-4 10 (40)

17, 20

Control - - - - +1.22 (D13)

Agent Median Median % of Regression Biosatitic Biological

AT/AC regression PR CR P value³ comments in % (day) (D24)

(D24)

. .... 20 - 4/8 2/8 ns Active huMAb2-3-SPDB-

DM4

42 - 0/8 0/8 ns Inactive anti- mu/huPD-L1/anti- 29 - 0/8 0/8 ns Active muCTLA-4 Agent Median Median % of Regression Biosatitic Biological

AT/AC regression PR CR P value³ comments in % (day) (D24)

(D24) huMAb2-3-SPDB- 20 100 (D39) 8/8 7/8 = 0.0321 Highly active

DM4

+ anti-mu/huPD- 14 100 (D39) 5/8 5/8 = 0.0014 Highly active

L1/anti-muCTLA-4

Control _ _ _ _

[0566]^a: Statistical analysis. The p-values were obtained using a contrast analysis to compare each treated group versus control using Bonferroni-Holm adjustment for multiplicity after a two-way Anova-Type with repeated measures on tumor volume changes from baseline. A probability less than 5% (p<0.05) was considered as significant. [0567] AT/AC = ratio of medians of tumor volume changes from baseline between treated and control groups; PR = Partial regression; CR = Complete regression.

Claims

CLAIMS]

1. A combination of (i) an antibody-drug conjugate (ADC) comprising an anti- CEACAM5 antibody and (ii) an anti-CTLA4 antibody, for use in the treatment of a cancer.

2. The combination for use according to claim 1 , wherein the anti-CEACAM5 antibody comprises a CDR-H1 having the amino acid sequence of SEQ ID NO: 1 , a CDR-H2 having the amino acid sequence of SEQ ID NO: 2, a CDR-H3 having the amino acid sequence of SEQ ID NO: 3, a CDR-L1 having the amino acid sequence of SEQ ID NO: 4, a CDR-L2 having the amino acid sequence NTR, and a CDR-L3 having the amino acid sequence of SEQ ID NO: 5.

3. The combination for use according to claim 1 or 2, wherein the antibody-drug conjugate comprises at least one chemotherapeutic agent.

4. The combination for use according to claim 3, wherein the chemotherapeutic agent is a maytansinoid selected from N2’-deacetyl-N2’-(3-mercapto-1 -oxopropyl)-maytansine (DM1 ), N2’-deacetyl-N2’-(4-methyl-4-mercapto-1 -oxopentyl)-maytansine (DM4), and combinations thereof.

5. The combination for use according to claim 3 or 4, wherein the anti-CEACAM5 antibody is covalently attached via non-cleavable linker to the at least one chemotherapeutic agent, the linker being selected from N-succinimidyl pyridyldithiobutyrate (SPDB), 4-(pyridin- 2-yldisulfanyl)-2-sulfo-butyric acid (sulfo-SPDB), and succinimidyl(N-maleimidomethyl) cyclohexane- 1 -carboxylate (SMCC).

6. The combination for use according to any one of claims 1 to 5, wherein the antibody-drug conjugate is tusamitamab ravtansine.

7. The combination for use according to any one of claims 1 to 6, wherein the antibody-drug conjugate is administered at a dose of 80 mg/m² to 210 mg/m², 80 mg/m² to 170 mg/m², or at a dose of 80 mg/m² to 150 mg/m², or at a dose of 80 mg/m² to 120 mg/m², or at a dose of 80 mg/m² to 100 mg/m².

8. The combination for use according to any one of claims 1 to 7, wherein the anti- CTLA4 antibody is ipilimumab, tremelimumab, quavonlimab, or zalifrelimab.

9. The combination for use according to any one of claims 1 to 8, wherein the anti- CTLA4 antibody is administered at a dose of 1 mg/kg to 10 mg/kg, or at a dose of 3 mg/kg to 10 mg/kg.

10. The combination for use according to any one of claims 1 to 9, wherein (i) the antibody-drug conjugate and (ii) the anti-CTLA4 antibody are administered on day 1 of a first cycle of treatment and on day 1 of at least one additional cycle(s) of treatment.

1 1 . The combination for use according to any one of claims 1 to 10, wherein (i) the antibody-drug conjugate and (ii) the anti-CTLA4 antibody are administered once every three weeks or once every two weeks.

12. The combination for use according to any one of claims 1 to 1 1 , further comprising (iii) an anti-PD-1 antibody or anti-PD-L1 antibody.

13. The combination for use according to claim 12, wherein the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, sintilimab, dostarlimab, and tislelizumab.

14. The combination for use according to claim 12 or 13, wherein the anti-PD-1 antibody or the anti-PD-L1 antibody is administered at a dose of 150 mg to 400 mg, or at a dose of 150 mg to 300 mg.

15. The combination for use according to any one of claims 1 to 14, wherein the cancer is a CEACAM5 positive cancer having a CEACAM5 immunohistochemical intensity > 2+ in > 1% and in < 50% of cancer cells or is a CEACAM5 positive cancer having a CEACAM5 immunohistochemical intensity > 2+ in > 50% of cancer cells.

16. The combination for use according to any one of claims 1 to 15, wherein the cancer is selected from hepatocellular carcinoma, colorectal cancer, gastric cancer, gastroesophageal junction adenocarcinoma (GEJ), esophageal cancer, lung cancer, uterine cervix cancer, pancreatic cancer, ovarian cancer, thyroid cancer, bladder cancer, endometrial cancer, breast cancer, liver cancer, biliary tract cancer, prostate cancer, neuroendocrine cancer, and skin cancer.

17. A kit-of-parts comprising (i) a pharmaceutical composition comprising an antibody-drug conjugate according to anyone of claims 1 to 7, and a pharmaceutically acceptable excipient, and (ii) a pharmaceutical composition comprising an anti-CTLA4 antibody and a pharmaceutically acceptable excipient, and, optionally, (iii) a pharmaceutical composition comprising an anti-PD-1 antibody or an anti-PD-L1 antibody and a pharmaceutically acceptable excipient.

18. The kit-of-parts according to claim 17, for use in the treatment of a cancer.

19. Use of a combination of (i) an antibody-drug conjugate according to anyone of claims 1 to 7, and (ii) an anti-CTLA4 antibody, and, optionally, (iii) an anti-PD-1 antibody or an anti-PD-L1 antibody, for the manufacture of a kit-of-parts, for the treatment of a cancer.