ENSAIO DE RNA MENSAGEIRO CEACAM5 PARA SELEÇÃO DE PACIENTES NA TERAPIA DO CÂNCER

Abstract

The disclosure relates to methods and uses for selecting and treating patients with cancer, where the cancer expresses CEACAM5. The methods include determining a log2- transformed, quantile-normalized, Transcripts Per Kilobase Million (TPM) value for CEACAM5 mRNA in an isolated sample of a tumor obtained from said subject, (ii) comparing said value with a reference value, and (iii) selecting said subject for a treatment of a cancer if the determined value is above the reference value. The agent for treating selected subjects can be tusamitamab ravtansine. Such agent can be used in combination with one or more additional agents to treat the cancer. In certain embodiments the cancer is non-squamous non- small cell lung cancer (NSQ NSCLC)

Description

[001] This application contains a Sequence Listing that was submitted electronically in .xml format and is incorporated herein by reference in its entirety. This .xml copy, created on March 12, 2024, is named PI022916.WO SANOFI (S327) LISTING.xml. [TECHNICAL FIELD]

[002] The present invention relates to the field of cancer therapy. [BACKGROUND OF THE TECHNIQUE]

[003] Despite recent advances in cancer treatment, a new effective treatment at the time of disease progression after first-line therapy remains necessary. Current therapeutic approaches for subsequent systemic options that combine an angiogenesis inhibitor with a systemic cytotoxic agent, such as docetaxel, involve serious hematological and other toxicities. Docetaxel, pemetrexed, or gemcitabine used as single cytotoxic agents offer very limited other options. Therefore, targeted cytotoxic therapies may offer an improvement in safety and tolerability, as well as efficacy.

[004] Lung cancer is the leading cause of cancer-related mortality globally [1]. In the United States, non-small cell non-squamous lung cancer (NSQ-NSCLC) comprises 45.4% of new lung cancer diagnoses and, among these, 47.7% have distant metastases [2] with an associated unsatisfactory five-year relative survival of 5-8% [3].

[005] Although some lung cancers respond to therapies targeting a variety of oncogene triggers [4] or to Petition 870250084766, dated 09 / 19 / 2025, page 10 / 149 2 / 115 immune checkpoint inhibitors (ICIs) [5], there is a continuing unmet need for innovative therapies for patients whose lung cancers are either untargetable or unresponsive to ICIs.

[006] Carcinoembryonic antigen-related cell adhesion molecules (CEACAMs) are cell surface glycoproteins involved in cell adhesion, cell signaling, and promoting cancer progression and metastasis [6]. Consequently, they are promising targets for new anticancer agents.

[007] Although different CEACAMs show specific expression patterns across tissues, they demonstrate similarity with respect to structural homology and amino acid sequences [7]. This requires that molecules targeting CEACAMs must bind specifically to individual CEACAMs to limit cross-reactivity and dose-limiting toxicity.

[008] CEACAM5 was first described in 1965 as a tumor-associated antigen in extracts of human colon cancer tissue (Gold P et al., J Exp Med. 1965;122(3):467-481). Since then, high levels of CEACAM5 expression have been observed in several epithelial tumors, while in normal adult tissue, its expression is limited to a few tissues (Hammarstrom S., Semin Cancer Biol. 1999;9(2):67-81; Thompson JA., Tumor Biol. 1995;16(1):10-16).

[009] Among CEACAMs, therapies targeting CEACAM5 are of particular interest in lung cancer. CEACAM5 is overexpressed in many epithelial tumors [8], which facilitates tumorigenesis and metastasis [9]. CEACAM5 is expressed at higher levels in NSCLC, but is not expressed in normal lung tissue [7, 10]. Furthermore, higher CEACAM5 expression in human NSCLC tissue is correlated with worse histological grade

[10] , and higher CEACAM5 expression is associated with survival in Petition 870250084766, dated 19 / 09 / 2025, p. 11 / 149 3 / 115 satisfactory in patients with NSCLC. [11, 12] Taken together, these findings suggest that CEACAM5 is a promising target for antibody-drug conjugate (ADC) therapy.

[0010] Antibody-drug conjugates (ADCs) have shown promise in improving outcomes in patients with lung cancer

[13] . Tusamitamab ravtansine (SAR408701) is a potential first-class ADC that selectively targets tumor cells expressing CEACAM5 without binding to CEACAM1, 6, or 8 glycoproteins [7]. It consists of a humanized CEACAM5 monoclonal antibody conjugated with a payload of the potent cytotoxic N2'-deacetyl-N-2'(4-methyl-4-mercapto-1-oxopentyl)-maitansine (DM4) maytansinoid via a cleavable ligand [7]. The tusamitamab ravtansine antibody component binds to the extracellular domain of CEACAM5, which is followed by internalization of the ADC and subsequent release of DM4 into the tumor cell

[14] . DM4 inhibits microtubule assembly, resulting in cell cycle arrest and apoptosis

[14] .Both DM4 and its active metabolite, S-methylDM4, cross the cell membrane, meaning that the cytotoxic effect of tusamitamab ravtansine is direct through specific binding to tumor cells expressing CEACAM5 and indirect through local diffusion of DM4 and S-methyl-DM4 (the observer effect)

[14] .

[0011] The promising preliminary antitumor activity of tusamitamab ravtansine in heavily pre-treated participants for NSQ NSCLC was demonstrated in an ongoing study (TED13751).

[0012] In the dose-escalation phase of a first Phase 1 / 2 human study of tusamitamab ravtansine among patients with advanced solid tumors for whom standard alternative therapy was not available (ClinicalTrials.gov NCT02187848), the primary dose-limiting toxicity was reversible keratopathy, and the maximum tolerated dose was 100 mg / m2 every 2 weeks.

[15] . Petition 870250084766, dated 09 / 19 / 2025, page 12 / 149 4 / 115

[0013] In the expansion phase of the same study among patients with advanced NSQ-NSCLC expressing high levels of CEACAM5, defined as at least 50% of tumor cells with 2+ or 3+ staining intensity on immunohistochemistry (IHC), tusamitamab ravtansine demonstrated a promising objective response rate of 20.3% and a favorable safety profile with the most common treatment-emergent adverse events being asthenia, reversible corneal events, peripheral neuropathy, dyspnea, and diarrhea, with infrequent hematological toxicities compared to those reported with docetaxel

[16] ; furthermore, among patients who achieved a partial response, 47% were treated for more than 1 year, suggesting that the response to tusamitamab ravtansine was durable and often sustained

[17] .

[0014] Although these clinical trial data appear promising, in clinical practice, potential barriers to pre-screening patients with cancer, for example, NSQ-NSCLC, who are likely to respond to CEACAM5-targeted therapies, include the availability of archival tumor biopsies for measuring CEACAM5 expression by immunohistochemistry.

[0015] Based on the Phase 1 / 2 study mentioned above, we explored biomarker associations with: 1) tumor expression of CEACAM5 by immunohistochemistry, with tumor CEACAM5 mRNA levels; and 2) whether CEACAM5 mRNA predicted the objective tumor response rate. [SUMMARY]

[0016] As shown in the examples, there was an enrichment of clinical responses in patients with a high level of CEACAM5 protein expression and being treated with tusamitamab ravtansine (tusamitamab ravtansine responders).

[0017] In the invention, the expression an enrichment of res Petition 870250084766, dated 09 / 19 / 2025, p. 13 / 149 5 / 115 clinical posts in patients in relation to treatment with tusamitamab ravtansine is intended to refer to an increase in the overall response rate (ORR) after administration of the given treatment. The ORR is defined as the proportion of patients who have a partial or complete response to therapy; it does not include stable disease and is a direct measure of the tumoricidal activity of drugs.

[0018] Furthermore, a correlation was found between the level of CEACAM5 protein expression in the tumor (measured by immunohistochemical staining (IHC)) and the level of tumor CEACAM5 mRNA. CEACAM5 mRNA expression was significantly downregulated in patients with high to moderate CEACAM5 protein expression. In addition, there was an enrichment of clinical responses in patients with high levels of tumor CEACAM5 mRNA and high immunohistochemical staining of CEACAM5 (IHC) (i.e., > intensity 2+ in > 50% of tumor cells) in responders to tusamitamab ravtansine.

[0019] Since an enrichment of clinical responses was shown with high expression of the CEACAM5 protein and an association between CEACAM5 protein expression and the level of CEACAM5 mRNA expression was shown, then the level of CEACAM5 mRNA expression was shown as a satisfactory biomarker, predictive of an enrichment of clinical responses, in patients to be treated with an anti-CEACAM5 antibody-drug conjugate, such as tusamitamab ravtasin; alternatively, the results show that patients can be selected based on the level of CEACAM5 mRNA instead of IHC.

[0020] The Example section therefore supports the use of CEACAM5 mRNA level as a biomarker to select and treat patients who need cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-Petition 870250084766, dated 09 / 19 / 2025, page 14 / 149 6 / 115 CEACAM5 conjugated with a cytotoxic agent.

[0021] In addition, the Example section supports the use of CEACAM5 mRNA level as a biomarker to select patients who need cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated with a cytotoxic agent for a further screening step with immunohistochemical staining of CEACAM5 (IHC) and subsequent treatment of patients who need cancer with the ADC.

[0022] It is common knowledge to a well-versed person that the level of protein expression does not necessarily correlate with the level of expression of the corresponding mRNA.

[0023] The invention is based, at least in part, on the observation that particular values ​​of CEACAM5 expression level, for example, CEACAM5 gene transcript level, for example, CEACAM5 mRNA level, for example, quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA may be useful as a biomarker for screening patients for treatment of cancers that typically express Cell Adhesion Molecule CEA 5 (CEACAM5) in their tumor cells.

[0024] In some embodiments, the present invention relates to a method for selecting an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, wherein the method comprises at least the following steps: (i) to determine a gene expression level value for CEACAM5 in an isolated tumor sample obtained from said individual, Petition 870250084766, dated 09 / 19 / 2025, page 15 / 149 7 / 115 (ii) compare said determined value with a reference value and (iii) select said individual for cancer treatment if the determined value is above the reference value.

[0025] Within the invention, an individual who needs the same is an individual who has cancer. An isolated sample of a tumor from said cancer can be used.

[0026] In this document, the terms individual, patient, individual who needs the same and patient who needs the same are used interchangeably.

[0027] In some embodiments, the present invention relates to a method for selecting an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, wherein the method comprises at least the following steps: (i) determine a gene expression level value for CEACAM5 in an isolated tumor sample obtained from said individual, (ii) compare said value determined in step (i) with a reference value for a CEACAM5 gene expression level, (iii) select said individual for a CEACAM5 immunohistochemical staining (IHC) test if the value determined in step (i) is above the reference value of step (ii), (iv) determine the intensity of a CEACAM5 protein expression level with said CEACAM5 immunohistochemical staining (IHC) test in an isolated tumor sample obtained from said individual, (v) compare said intensity determined in step (iv) with a reference intensity, and. Petition 870250084766, dated 09 / 19 / 2025, page 16 / 149 8 / 115 (vi) select said individual for cancer treatment if the intensity determined in step (iv) is above the reference intensity.

[0028] The tumor samples from step (i) and step (iv) may be the same or different samples.

[0029] In some embodiments, the present invention relates to a method for selecting and treating an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, wherein the method comprises at least the steps of: (i) to determine a gene expression level of CEACAM5 in an isolated tumor sample obtained from said individual, (ii) to compare said determined value with a reference value, (iii) to select said individual for cancer treatment if the determined value is above the reference value, and (iv) to administer to said selected individual an effective amount of said ADC.

[0030] In some embodiments, the present invention relates to a method for selecting and treating an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, wherein the method comprises at least the steps of: (i) to determine a value for the gene expression level of CEACAM5 in an isolated tumor sample obtained from said individual, (ii) to compare said value determined in step (i) with a Petition 870250084766, dated 09 / 19 / 2025, page 17 / 149 9 / 115 reference value of a CEACAM5 gene expression level, (iii) select said individual for a CEACAM5 immunohistochemical staining (IHC) test if the value determined in step (i) is above the reference value of step (ii), (iv) determine an intensity of a CEACAM5 protein expression level with said CEACAM5 immunohistochemical staining (IHC) test in an isolated tumor sample obtained from said individual, (v) compare said intensity determined in step (iv) with a reference intensity, e. (vi) select said individual for cancer treatment if the intensity determined in step (iv) is above the reference intensity, and (vii) administer to said selected individual an effective amount of said ADC.

[0031] In some embodiments, the present invention relates to an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, for use in the treatment of cancer in an individual in need thereof, the use comprising: (i) to determine a gene expression level of CEACAM5 in an isolated tumor sample obtained from said individual, (ii) to compare said determined value with a reference value and (iii) to administer to said individual an effective amount of said ADC if said determined value is above the reference value.

[0032] In some embodiments, the present invention relates to an antibody-drug conjugate (ADC) comprising an antibody Petition 870250084766, dated 09 / 19 / 2025, page 18 / 149 10 / 115 anti-CEACAM5 conjugated to a cytotoxic agent, for use in the treatment of cancer in an individual in need thereof, the use comprises: (i) determine a gene expression level value for CEACAM5 in an isolated tumor sample obtained from said individual, (ii) compare said value determined in step (i) with a reference value for a CEACAM5 gene expression level, (iii) select said individual for a CEACAM5 immunohistochemical staining (IHC) test if the value determined in step (i) is above the reference value of step (ii), (iv) determine the intensity of a CEACAM5 protein expression level with said CEACAM5 immunohistochemical staining (IHC) test in an isolated tumor sample obtained from said individual, (v) compare said intensity determined in step (iv) with a reference intensity, and. (vi) administer to said individual an effective amount of said DCA if said determined intensity is above the reference intensity.

[0033] In some embodiments, the present invention relates to the use of a measurement of a CEACAM5 gene expression level value in an isolated tumor sample obtained from an individual in need thereof to characterize said tumor as a CEACAM5-high expressing tumor.

[0034] In some embodiments, the present invention relates to the use of a measurement of a value of the expression level of the CEACAM5 gene in an isolated tumor sample obtained from an individual in need thereof in order to select said individual for an immunohistochemical staining test for CEACAM5 (IHC). Petition 870250084766, dated 09 / 19 / 2025, p. 19 / 149 11 / 115

[0035] In some embodiments, the present invention relates to the use of a measurement of the expression level of the CEACAM5 gene in an isolated tumor sample obtained from an individual in need thereof in order to select said individual for cancer treatment with a CEACAM5-targeting therapeutic agent, for example, an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent.

[0036] In some embodiments, the present invention relates to the use of a measurement of the expression level of the CEACAM5 gene in an isolated tumor sample obtained from an individual in need thereof in order to select said individual for a CEACAM5 immunohistochemical staining test (IHC), said staining test being for selecting said individual for cancer treatment with a CEACAM5-targeting therapeutic agent, for example, an antibody-drug conjugate comprising an anti-CEACAM5 antibody conjugated with a cytotoxic agent.

[0037] In some modalities, the value of the CEACAM5 gene expression level may be a measure of a CEACAM5 gene transcript.

[0038] In some embodiments, the CEACAM5 gene transcript may be an mRNA.

[0039] In some embodiments, the methods and uses of the invention comprise a step of determining a value of a CEACAM5 mRNA level in an isolated tumor sample.

[0040] According to one of its objectives, the present invention relates to a method for selecting an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugate. Petition 870250084766, dated 09 / 19 / 2025, page 20 / 149 12 / 115 cattle with a cytotoxic agent, wherein the method comprises at least the following steps: (i) determine a quantile-normalized, log2-transformed Transcripts Per Million Kilobases (TPM) value for CEACAM5 mRNA in a single tumor sample obtained from said individual, (ii) compare said value with a reference value, and (iii) select said individual for cancer treatment if the determined value is above the reference value.

[0041] In some modalities, the value of the CEACAM5 gene expression level may be a measure of a CEACAM5 gene transcript.

[0042] In some embodiments, the CEACAM5 gene transcript may be an mRNA.

[0043] According to one of its objectives, the present invention relates to a method for selecting an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, wherein the method comprises at least the following steps: (i) determine a quantile-normalized, log2-transformed Transcripts Per Million Kilobases (TPM) value for CEACAM5 mRNA in a single tumor sample obtained from said individual, (ii) compare said value determined in step (i) with a reference value, (iii) select said individual for a CEACAM5 immunohistochemistry (IHC) staining test if the value determined in step (i) is above the reference value from step (ii), (iv) determine an intensity of an expression level. Petition 870250084766, dated 09 / 19 / 2025, page 21 / 149 13 / 115 of the CEACAM5 protein with said CEACAM5 immunohistochemical staining (IHC) test in an isolated tumor sample obtained from said individual, (v) compare said intensity determined in step (iv) with a reference intensity, and. (vi) select said individual for cancer treatment if the determined intensity is above the reference intensity.

[0044] According to another of its objectives, the present invention relates to a method for selecting and treating an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated with a cytotoxic agent, wherein the method comprises at least the steps of: (i) determine a quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA in a single tumor sample obtained from said individual, (ii) compare said value with a reference value, (iii) select said individual for cancer treatment if the determined value is above the reference value, and (iv) administer to said selected individual an effective amount of said ADC.

[0045] Thus, cancer can be treated.

[0046] According to another of its objectives, the present invention relates to a method for selecting and treating an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated with a cytotoxic agent, wherein the method comprises at least the steps of: Petition 870250084766, dated 09 / 19 / 2025, page 22 / 149 14 / 115 (i) determine a quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA in a single tumor sample obtained from said individual, (ii) compare said value determined in step (i) with a reference value, (iii) select said individual for a CEACAM5 immunohistochemical staining (IHC) test if the value determined in step (i) is above the reference value of step (ii), (iv) determine an intensity of a CEACAM5 protein expression level with said CEACAM5 immunohistochemical staining (IHC) test in a single tumor sample obtained from said individual, (v) compare said intensity determined in step (iv) with a reference intensity, and. (vi) select said individual for cancer treatment if the determined intensity is above the reference intensity, and (vii) administer to said selected individual an effective amount of said ADC.

[0047] According to another of its objectives, the present invention relates to an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, for use in the treatment of cancer in an individual in need thereof, the use comprising (i) determining a quantile-normalized log2-transformed Transcripts Per Million Kilobases (TPM) value for CEACAM5 mRNA in an isolated tumor sample obtained from said individual, (ii) comparing said value with a reference value, and (iii) administering to said individual a quantity Petition 870250084766, dated 09 / 19 / 2025, page 23 / 149 15 / 115 effective of said ADC if the determined value is above the reference value.

[0048] In accordance with another of its objectives, the present invention relates to an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, for use in the treatment of cancer in an individual in need thereof, the use comprising: (i) determine a quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA in a single tumor sample obtained from said individual, (ii) compare said value determined in step (i) with a reference value, (iii) select said individual for a CEACAM5 immunohistochemical staining (IHC) test if the value determined in step (i) is above the reference value of step (ii), (iv) determine an intensity of a CEACAM5 protein expression level with said CEACAM5 immunohistochemical staining (IHC) test in a single tumor sample obtained from said individual, (v) compare said intensity determined in step (iv) with a reference intensity, and. (vi) administer to said individual an effective amount of said DCA if the determined intensity is above the reference intensity.

[0049] As shown in the Examples section, using ribonucleic acid (RNA) sequencing, the expression of approximately 15,000 genes, including CEACAM5, was evaluated. RNA data were filtered and defined as quantile-normalized transcripts. Petition 870250084766, dated 09 / 19 / 2025, page 24 / 149 16 / 115 log per million kilobases (TPM). Differential gene expression analysis identified CEACAM5 mRNA as the gene most strongly associated (and the only significant adjusted P = 0.00265) with high vs. moderate CEACAM5 expression by IHC. CEACAM5 mRNA transcripts per million are elevated in high vs. moderate CEACAM5 expression; and (C) CEACAM5 mRNA correlates with CEACAM5 IHC H-score. Consequently, CEACAM5 gene expression level (RNA) can be used to identify and select patients responsive to CEACAM5-targeted treatment, such as an antibody-drug conjugate comprising an anti-CEACAM5 antibody conjugated with a cytotoxic agent.

[0050] In the invention, the antibody-drug conjugate comprising an anti-CEACAM5 antibody conjugated with a cytotoxic agent, antibody-drug conjugate and ADC are used interchangeably.

[0051] In some modes, the ADC can be used in an effective quantity.

[0052] In some forms, the use may be in an individual who needs it.

[0053] In an application as disclosed in this document, the mRNA biomarker CEACAM5 is measured in an isolated biological sample.

[0054] In some embodiments, the anti-CEACAM5 antibody may comprise an HCDR1 having the amino acid sequence of SEQ ID NO: 1, an HCDR2 having the amino acid sequence of SEQ ID NO: 2, an HCDR3 having the amino acid sequence of SEQ ID NO: 3, an LCDR1 having the amino acid sequence of SEQ ID NO: 4, an LCDR2 having the amino acid sequence of NTR and an LCDR3 having the amino acid sequence of SEQ ID NO: 5. Petition 870250084766, dated 09 / 19 / 2025, p. 25 / 149 17 / 115

[0055] In some embodiments, the anti-CEACAM5 antibody may comprise a variable heavy chain (VH) domain consisting of SEQ ID NO: 6 and a variable light chain (VL) domain consisting of SEQ ID NO: 7.

[0056] In some embodiments, the anti-CEACAM5 antibody may be tusamitamab.

[0057] In some embodiments, the cytotoxic agent may be selected from the group consisting of radioisotopes, protein toxins, small molecule toxins, and any combination thereof.

[0058] In some embodiments, the small molecule toxin may be selected from among antimetabolites, DNA alkylating agents, DNA crosslinking agents, DNA intercalating agents, antimicrotubular agents, topoisomerase inhibitors and any combination thereof.

[0059] In some embodiments, the antimicrotubule agent may be selected from taxanes, vinca alkaloids, maytansinoids, colchicine, podophyllotoxin, griseofulvin, and any combination thereof.

[0060] In some embodiments, the cytotoxic agent may be a maytansinoid or maytansinoid analogue.

[0061] In some embodiments, the maytansinoid may be selected from N2'-deacetyl-N2'-(3-mercapto-1-oxopropyl)-maytansine (DM1) or N2'-deacetyl-N2'-(4-methyl-4-mercapto-1-oxopentyl)-maytansine (DM4) and combinations thereof.

[0062] In some embodiments, the anti-CEACAM5 antibody can be covalently linked via a cleavable or non-cleavable ligand to at least one chemotherapeutic agent.

[0063] In some embodiments, the ligand may be selected from N-succinimidyl pyridyldithiobutyrate (SPDB), 4-(pyridin-2 Petition 870250084766, dated 09 / 19 / 2025, page 26 / 149 18 / 115 yldisulfanyl)-2-sulfo-butyric acid (sulfo-SPDB) and succinimidyl (N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC).

[0064] In some embodiments, the anti-CEACAM5 antibody may be covalently linked to a ligand cleavable to at least one chemotherapeutic agent, wherein said ligand is selected from N-succinimidyl pyridyldithiobutyrate (SPDB), 4-(pyridin-2-yldisulfanyl)-2-sulfobutyric acid (sulfo-SPDB) and succinimidyl(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC).

[0065] In some embodiments, the CEACAM5-antibody may comprise a heavy chain (VH) consisting of SEQ ID NO: 8 and a light chain (VL) consisting of SEQ ID NO: 9 (huMAb2-3), which may be covalently linked to N2'-deacetyl-N-2'(4-methyl-4-mercapto-1-oxopentyl)-maitansine (DM4) via succinimidyl pyridyldibutyrate (SPDB).

[0066] In some embodiments, the antibody-drug conjugate may be characterized by a drug-antibody ratio (DAR) ranging from 1 to 10.

[0067] In some embodiments, the antibody-drug conjugate may be tusamitamab ravtansine.

[0068] In some embodiments, the reference value may be a quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA.

[0069] In some modalities, the reference value may be at least about 7 to about 13.

[0070] In some embodiments, the preference value may be at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, at least about 12 or at least about 13.

[0071] In some modalities, quantile normalization can be achieved by (i) classifying sample transcripts by level of ex Petition 870250084766, dated 09 / 19 / 2025, page 27 / 149 19 / 115 pressure, (ii) calculate an average value for genes that occupy the same class and (iii) replace the values ​​of all genes that occupy the same class with this average value.

[0072] In some modes, the expression level of transcripts can be measured in Fragments per Million Kilobases (FPKM) before being converted to TPM.

[0073] In some forms, Fragments per Million Kilobases (FPKM) can be obtained by counting the total number of transcripts in the sample, dividing the transcript counts obtained by 1,000,000, and dividing the values ​​obtained by the gene length, in kilobases.

[0074] In some modalities, the cancer may be selected from a group consisting of hepatocellular carcinoma, colorectal cancer, gastric cancer, adenocarcinoma of the gastroesophageal junction (GEJ), esophageal cancer, lung cancer, cervical 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.

[0075] In certain modalities, cancer may be selected from the group consisting of colorectal cancer, gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, esophageal cancer, lung cancer, cervical 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.

[0076] In some modalities, the cancer may be selected from among colorectal cancer, gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, esophageal cancer, pancreatic cancer, and lung cancer. Petition 870250084766, dated 09 / 19 / 2025, page 28 / 149 20 / 115

[0077] In some forms, the cancer may be gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, or esophageal cancer.

[0078] In some forms, the cancer may be gastric cancer.

[0079] In some forms, the cancer may be colorectal cancer.

[0080] In some forms, the cancer may be pancreatic cancer.

[0081] In some forms, the cancer may be adenocarcinoma of the gastroesophageal junction (GEJ).

[0082] In some forms, the cancer may be lung cancer.

[0083] In some forms, lung cancer may be non-small cell squamous lung cancer (NSCLC).

[0084] In some forms, non-small cell non-squamous lung cancer may be an advanced or metastatic NSCLC SCN.

[0085] In some modalities, non-small cell non-squamous lung cancer does not have a sensitizing mutation of the epidermal growth factor receptor (EGFR) or a mutation of the B1 homolog of the murine sarcoma virus v-raf (BRAF) oncogene or alterations in anaplastic lymphoma kinase / c-ros 1 (ALK / ROS) oncogene.

[0086] In some modalities, the anti-CEACAM5 antibody may be tusamitamab.

[0087] In some modalities, the ADC may be tusamitamab ravtansine.

[0088] In some modalities, ADC can be administered at a dose of > 80 mg / m2, relative to body surface area. Petition 870250084766, dated 09 / 19 / 2025, p. 29 / 149 21 / 115 of said individual, approximately once every two weeks, or the ADC may be administered at a dose of > 80 mg / m2, relative to the body surface area of ​​said individual, approximately once every three weeks.

[0089] In some embodiments, ADC 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.

[0090] In some modalities, ADC can be administered at a dose of 80, 100, 120, 150, 170, 180 or 210 mg / m2.

[0091] In some embodiments, the method or use as described herein further comprises administering to the individual an effective amount of at least one additional agent effective for treating cancer.

[0092] In some embodiments, the additional agent may be selected from the group consisting of an immune checkpoint inhibitor (ICI), a platinum-based chemotherapy, pemetrexed, anti-VEGFR2, FOLFOX, FOLFIRI, TAS-102, anti-EGFR and any combination thereof.

[0093] In some embodiments, the ICI may be an anti-PD-1 antibody or an anti-PD-L1 antibody.

[0094] In some embodiments, the anti-PD-1 antibody may be selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, sintilimab, dostarlimab and tislelizumab.

[0095] In some embodiments, the anti-PD-L1 antibody may be selected from the group consisting of atezolizumab, avelumab and durvalumab.

[0096] In some modalities, the individual receives an effective amount of tusamitamab ravtansine and pembrolizumab.

[0097] In some modalities, the method or use as described Petition 870250084766, dated 09 / 19 / 2025, page 30 / 149 22 / 115 written here may additionally include administering an effective amount of platinum-based chemotherapy to the individual.

[0098] In some modalities, platinum-based chemotherapy may be selected from cisplatin and carboplatin.

[0099] In some embodiments, the method or use as described herein may additionally comprise administering an effective amount of pemetrexed to the individual. [DESCRIPTION OF THE FIGURES]

[00100] Figure 1: shows that CEACAM5 mRNA transcripts per million are elevated in high vs. moderate expression of CEACAM5.

[00101] Figure 2: shows correlation of CEACAM5 mRNA with CEACAM5 IHC H-score.

[00102] Figure 3: This shows the correlation between CEACAM5 mRNA level and CEACAM5 expression by immunohistochemistry, recorded as the sum of the percentage of tumor cells expressing target intensity at least 2+ according to the patient's response to treatment with tusamitamab ravtansine.

[00103] Figure 4: This shows the correlation between CEACAM5 mRNA level and CEACAM5 expression by immunohistochemistry, recorded as the sum of the percentage of tumor cells expressing target intensity at least 2+ in patients who responded to treatment with tusamitamab ravtansine.

[00104] Figure 5: Shows box plots of the expression of CEACAM family members according to clinical responses in patients treated with tusamitamab ravtansine. [BRIEF DESCRIPTION OF THE SEQUENCES]

[00105] SEQ ID NO: 1-5 shows the CDR-H1, CDR-H2, CDR-H3, CDR-L1 and CDR-L3 sequences of the anti-CEACAM5 antibody (huMAb2-3).

[00106] SEQ ID NO: 6 shows the variable domain sequence of Petition 870250084766, dated 09 / 19 / 2025, page 31 / 149 23 / 115 heavy chain (VH) of the anti-CEACAM5 antibody (huMAb2-3).

[00107] SEQ ID NO: 7 shows the sequence of the variable domain of the light chain (VL) of the anti-CEACAM5 antibody (huMAb2-3).

[00108] SEQ ID NO: 8 shows the heavy chain sequence of the anti-CEACAM5 antibody (huMAb2-3).

[00109] SEQ ID NO: 9 shows the light chain sequence of the anti-CEACAM5 antibody (huMAb2-3). [DETAILED DESCRIPTION] Definitions

[00110] Unless otherwise defined herein, the scientific and technical terms used in combination with the present invention shall have the meanings that are commonly understood by those of common skill in the art. For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, PeiShow, 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 the skilled with a general dictionary of many of the terms used in the present invention. Exemplary methods and materials are described below, although methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. In case of conflict, this descriptive report, including definitions, shall prevail.In general, the 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 in this document is that which is well known and commonly used in the art. Enzymatic reactions and purification techniques are performed in accordance with the aforementioned. Petition 870250084766, dated 09 / 19 / 2025, page 32 / 149 24 / 115 Manufacturer specifications, as commonly achieved in the art or as described herein. Additionally, unless otherwise required by the context, singular terms should include plurals and plural terms should include the singular.

[00111] Units, prefixes and symbols are denoted in their form accepted by the International System of Units (Système International des Unités (SI)). Numerical ranges are inclusive of the numbers that define the range. Unless otherwise indicated, amino acid sequences are written from left to right in the amino-to-carboxy orientation. The headings provided in this document are not limitations of the various aspects of the invention. Consequently, the terms defined immediately below are defined more fully by way of reference to the descriptive report in its entirety.

[00112] All publications and other references mentioned in this document are incorporated by reference in their entirety. Although a number of documents are cited in this document, such citation does not constitute an admission that any of these documents forms part of the common general knowledge in the art.

[00113] Throughout this descriptive report and modalities, the words have and comprise, or variations such as has, having, comprises or comprising, will be understood as implying the inclusion of a given integer or group of integers, but not the exclusion of any other integer or group of integers. It is understood that, whenever aspects are described herein with the language comprising, analogous aspects described in terms of consisting of and / or consisting essentially of are also provided. Petition 870250084766, dated 09 / 19 / 2025, page 33 / 149 25 / 115

[00114] It should be noted that the term "an" or "an entities" refers to one or more of these entities; for example, a nucleotide sequence is understood as representing one or more nucleotide sequences. Thus, the terms "an," "one or more," and "at least one" may be used interchangeably in this document.

[00115] Furthermore, and / or where used herein shall be understood as a specific invention 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 in this document, includes A and B, A or B, A (alone) and B (alone). Similarly, 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).

[00116] The term approximately or about is used here to mean approximately, roughly, around, or within the zones of. When the term about is used together with a numerical range, it modifies that range by extending the limits above and below the established numerical values. In general, the term about can modify a numerical value above and below the indicated value by a variation of, for example, 10 percent, upwards or downwards (higher or lower). In some modalities, the term indicates a deviation from the indicated numerical value of ± 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 modalities, approximately indicates a deviation from the indicated numerical value of ± 10%. In some modalities, approximately indicates a deviation from the indicated numerical value of ± 5%. In some modalities, approximately indicates a deviation from the indicated numerical value of ± 4%. Petition 870250084766, dated 09 / 19 / 2025, p. 34 / 149 26 / 115 In some modalities, approximately indicates a deviation from the indicated numerical value of ± 3%. In some modalities, approximately indicates a deviation from the indicated numerical value of ± 2%. In some modalities, approximately indicates a deviation from the indicated numerical value of ± 1%. In some modalities, approximately indicates a deviation from the indicated numerical value of ± 0.9%. In some modalities, approximately indicates a deviation from the indicated numerical value of ± 0.8%. In some modalities, approximately indicates a deviation from the indicated numerical value of ± 0.7%. In some modalities, approximately indicates a deviation from the indicated numerical value of ± 0.6%. In some modalities, approximately indicates a deviation from the indicated numerical value of ± 0.5%. In some modalities, approximately indicates a deviation from the indicated numerical value of ± 0.4%. In some modalities, approximately indicates a deviation from the indicated numerical value of ± 0.3%.In some modalities, approximately indicates a deviation from the indicated numerical value of ± 0.1%. In some modalities, approximately indicates a deviation from the indicated numerical value of ± 0.05%. In some modalities, approximately indicates a deviation from the indicated numerical value of ± 0.01%.

[00117] An antibody can be a natural or conventional antibody in which two heavy chains are linked together 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 classes (or isotypes) of heavy chain that 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 Petition 870250084766, dated 09 / 19 / 2025, page 35 / 149 27 / 115 Variables of both light (VL) and heavy (VH) chains determine antigen binding recognition and specificity. The constant region domains of light (CL) and heavy (CH) chains confer important biological properties such as antibody chain association, secretion, transplacental mobility, complementary binding, and binding to Fc receptors (FcR). The Fv fragment is the N-terminal portion of the Fab fragment of an immunoglobulin and consists of the variable portions of a light chain and a heavy chain. Antibody specificity resides in the structural complementarity between the antibody combining site and the antigenic determinant. Antibody combining sites are composed of residues that are primarily hypervariable regions or complementarity determinants (CDRs). Occasionally, residues from non-hypervariable regions or framework (FR) regions influence the overall domain structure and, consequently, the combining site.Complementarity Determinant Regions, or CDRs, therefore refer to amino acid sequences that 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. The antigen-binding site of a conventional antibody therefore includes six CDRs comprising the CDR defined from each of the heavy chain V region and the light chain V region.

[00118] As used in this document, the term antibody is intended to refer to conventional antibodies and fragments thereof, as well as single-domain antibodies and fragments thereof, in particular, variable heavy chain single-domain antibodies and chimeric, humanized, bispecific or multispecific antibodies. Antibody fragments considered in this Petition 870250084766, dated 09 / 19 / 2025, p. 36 / 149 28 / 115 document are antigen-binding fragments.

[00119] Framework regions (FRs) refer to amino acid sequences arranged between CDRs, that is, those portions of variable regions of immunoglobulin light and heavy chains 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.

[00120] In the context of the invention, the definition of CDR / FR in an immunoglobulin light or heavy chain must be determined based on the definition of IMGT (Lefranc et al. Dev. Comp. Immunol., 2003, 27(1):55-77; www.imgt.org).

[00121] As used in this document, antibody or immunoglobulin also includes single-domain antibodies, which have been described more recently and are antibodies whose complementarity-determining regions are part of a single-domain polypeptide. Examples of single-domain antibodies include heavy-chain antibodies, naturally occurring light-chain-deficient antibodies, single-domain antibodies derived from conventional four-chain antibodies, and engineered single-domain antibodies. Single-domain antibodies can be derived from any species, including but not limited to mice, humans, camels, llamas, goats, rabbits, and cattle. Single-domain antibodies can be naturally occurring single-domain antibodies known as light-chain-deficient heavy-chain antibodies. In particular, camelid species, for example, camels, dromedaries, Petition 870250084766, dated 09 / 19 / 2025, page 37 / 149 29 / 115 Llamas, alpacas, and guanacos produce heavy-chain antibodies that are naturally devoid of a light chain. Camelid heavy-chain antibodies also lack a CH1 domain.

[00122] The variable heavy chain of these single-domain antibodies devoid of light chains is known in the art as VHH or Nanobody®. Similar to conventional VH domains, VHHs contain four RFs and three CDRs. VHH has 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 a high yield. Additionally, VHH is very stable and resistant to protease action. The properties and production of VHH were reviewed by Harmsen and De Haard HJ (Appl. Microbiol. Biotechnol. 2007 Nov;77(1):13-22).

[00123] The term monoclonal antibody or mAb as used in this document refers to an antibody molecule of a single amino acid sequence that is directed against a specific antigen, and should not be interpreted as requiring the production of the antibody by any specific method. A monoclonal antibody can be produced by a single clone of B cells or hybridoma, but it can also be recombinant, i.e., produced by protein manipulation.

[00124] The term humanized antibody refers to an antibody that is wholly or partially of non-human origin, and that 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 often human CH and CL domains.

[00125] Antibody fragments (conventional) comprise Petition 870250084766, dated 09 / 19 / 2025, p. 38 / 149 30 / 115 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 can also be a single-domain antibody, such as a heavy chain or VHH antibody.

[00126] The term Fab denotes an antibody fragment that has a molecular weight of about 50,000 and antigen-binding activity in which about half of the N-terminal side of the heavy chain and the entire light chain are linked together via a disulfide bond. It is usually obtained between fragments via IgG with a protease, such as papain.

[00127] The term F(ab')2 refers to an antibody fragment that has a molecular weight of approximately 100,000 and antigen-binding activity that is slightly higher than two identical Fab fragments through a disulfide linkage in the hinge region. It is usually obtained between fragments by treating IgG with a protease, such as pepsin.

[00128] The term Fab' refers to an antibody fragment that has a molecular weight of about 50000 and antigen-binding activity that is obtained by cutting a disulfide bond from the hinge region of F(ab')2.

[00129] A single-chain Fv polypeptide (scFv) is a covalently linked VH::VL heterodimer that is usually expressed from a gene fusion including genes encoding VH and VL linked by a peptide-encoding linker. The human scFv fragment of the invention includes CDRs that are maintained in the appropriate conformation, in particular, through the use of gene recombination techniques. Divalent and multivalent antibody fragments can Petition 870250084766, dated 09 / 19 / 2025, page 39 / 149 31 / 115 can form spontaneously by the association of monovalent scFvs, or they can be generated through the coupling of monovalent scFvs by a peptide ligand, such as divalent sc(Fv)2. dsFv is a VH::VL heterodimer stabilized by a disulfide bond. (dsFv)2 denotes two dsFvs coupled by a peptide ligand.

[00130] The term bispecific antibody or BsAb denotes an antibody that combines the antigen-binding sites of two antibodies within a single molecule. Thus, BsAbs have the ability to bind simultaneously to two different antigens. Genetic manipulation has been used with increasing frequency to design, modify, and produce antibodies or antibody derivatives with a desired set of binding properties and effector function as described, for example, in document no. EP 2 050 764 A1.

[00131] The term multispecific antibody denotes an antibody that combines the antigen-binding sites of two or more antibodies within a single molecule.

[00132] The term diabodies refers to small antibody fragments with two antigen-binding sites whose fragments comprise a variable heavy chain (VH) domain connected to a variable light chain (VL) domain on the same polypeptide chain (VH-VL). Through the use of 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 complementarity domains of another chain and create two antigen-binding sites.

[00133] An amino acid sequence that is at least 85% identical to a reference sequence is a sequence that has, throughout its 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.

[00134] A percentage of sequence identity between if Petition 870250084766, dated 09 / 19 / 2025, page 40 / 149 32 / 115 Amino acid sequences can be determined by comparing two sequences, ideally aligned in a comparison window, where the portion of the polynucleotide or polypeptide sequence in the comparison window may contain additions or deletions (i.e., gaps) when compared to the reference sequence (which does not contain additions or deletions) for an ideal fragment of the two sequences. The percentage is calculated by determining the number of positions where the identical nucleic acid base or amino acid residue occurs in both sequences to yield the number of matching positions, dividing the number of matching positions by the total number of positions in the comparison window, and multiplying the result by 100 to yield the percentage of sequence identity. Ideal sequence alignment for comparison is conducted by global pairwise alignment, for example, using the Needleman and Wunsch J. Mol. Biol. algorithm.48:443 (1970). The sequence identity percentage can be readily determined, for example, using the Needle program with the BLOSUM62 matrix, and the following parameters: gap opening = 10, gap length = 0.5.

[00135] A conservative amino acid substitution is one in which an amino acid residue is replaced by another amino acid residue that has 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 alter the functional properties of a protein. Examples of amino acid groups 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 Petition 870250084766, dated 09 / 19 / 2025, p. 41 / 149 33 / 115: tyrosine and tryptophan; 5) basic side chains: lysine, arginine, and histidine; 6) acidic side chains: aspartic acid and glutamic acid; and 7) sulfur-containing side chains: cysteine ​​and methionine. Conservative amino acid substitution groups can also be defined based on amino acid size.

[00136] By purified and isolated, it is understood, when referring to a polypeptide (i.e., the antibody of the invention) or a nucleotide sequence in which the indicated molecule is present, that there is a substantial absence of other biological macromolecules of the same type. The term purified as used in this particular document means that at least 75%, 85%, 95% or 98% by weight of biological macromolecules of the same type are present. An isolated nucleic acid molecule encoding a particular polypeptide refers to a nucleic acid molecule that is substantially free of other nucleic acid molecules that do not encode the polypeptide in question; however, the molecule may include some additional bases or moieties that do not detrimentally affect the basic characteristics of the composition.

[00137] As used in this document, the term individual or patient denotes a mammal, such as a rodent, a feline, a canine, and a primate. In particular, an individual according to the invention is a human being.

[00138] To administer or administration, as used herein, refers to the delivery to an individual of a composition described herein. The composition may be administered to an individual using methods known in the art. In particular, the composition may be administered intravenously, subcutaneously, intramuscularly, intradermally or through any mucous surface, for example, orally, sublingually, buccally, nasally, rectally, vaginally or pulmonaryly. In some embodiments, administration is intravenous. In some mo Petition 870250084766, dated 09 / 19 / 2025, p. 42 / 149 34 / 115 doses, the administration is subcutaneous.

[00139] The terms treat or treatment or therapy refer to the administration of a compound or composition according to the invention with the aim of curing, healing, relieving, alleviating, altering, remedying, improving or affecting a disorder, the symptoms of the condition or to prevent or delay the onset of symptoms, complications or otherwise interrupt or inhibit the further development of the disorder in a statistically significant manner. More particularly, treat or treatment includes any approach to obtain beneficial or desirable results in an individual's cancer condition.Beneficial or desired clinical outcomes may include, but are not limited to, relief or improvement of one or more cancer symptoms or conditions, decrease or reduction in the extent of a cancer disease or cancer symptom, stabilization (i.e., non-aggravation) of the state of a cancer disease or cancer symptom, prevention of a cancer disease or the spread of a cancer symptom, delay or retardation of cancer disease or cancer symptom progression, complete improvement or palliation of cancer disease, reduction in cancer disease recurrence, and partial or complete remission, whether detectable or undetectable. In other words, treatment, as used herein, includes any cure, improvement, or reduction of a cancer disease or symptom. A reduction of a symptom or disease means a decrease in the severity or frequency of the disease or symptom, or elimination of the disease or symptom.

[00140] As used in this document, the term effective amount refers to an amount that provides a therapeutic benefit in the treatment, prevention, or management of considered pathological processes. The specific amount that is therapeutically effective can be readily determined by a physician. Petition 870250084766, dated 09 / 19 / 2025, page 43 / 149 35 / 115 mmHg 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.

[00141] The unit mg / m2 indicates the amount of compound in mg per m2 of body surface area of ​​an individual administered per dose. A person skilled in the art is aware of how to determine the required amount of compound for the individual to be treated based on their body surface area, which, in turn, can be calculated based on height and body weight.

[00142] The unit mg / kg indicates the amount of compound in mg per kg of patient body weight administered per dose. A person skilled in the art is aware of how to determine the required amount of compound for the individual to be treated based on their body weight.

[00143] It is appreciated that certain features of the invention, which are, for the sake of clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, several features of the invention, which are, for the sake of brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination.

[00144] Biomarker is intended to refer to a biological molecule, for example, a protein or a metabolite, that is differentially present, increased or decreased, in a biological sample obtained from an individual or group of individuals having a first phenotype, for example, having a disease such as cancer, compared with a biological sample from an individual or group of individuals having a second phenotype, for example, not having the disease. In use, the biomarker is isolated from the individual.

[00145] Sample or biological sample is intended to refer to Petition 870250084766, dated 09 / 19 / 2025, page 44 / 149 36 / 115 a biological material isolated from an individual. The biological sample may contain any biological material suitable for detecting the biomarker, i.e., CEACAM5 mRNA, and may comprise cellular and / or non-cellular material from the individual. The sample may be isolated from any suitable biological tissue or fluid, such as, for example, kidney tissue, blood, blood plasma (plasma), blood serum (serum), urine, or cerebrospinal fluid (CSF). In some embodiments, a biological sample is a sample of blood plasma (plasma) or blood serum (serum).

[00146] A reference value or threshold value is intended to refer to a CEACAM5 mRNA level that is indicative of a particular disease state, phenotype, such as cancer, or lack thereof, as well as combinations of disease states, phenotypes, or lack thereof, in the individual in question.

[00147] Unless otherwise defined, all technical and scientific terms used in this document have the same meaning as commonly understood by a person of ordinary skill in the art to which this invention pertains. Although any methods and materials similar or equivalent to those described in this document may also be used in the practice or testing of the present invention. All publications mentioned in this document are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited.

[00148] The list of sources, ingredients and components as described herein are listed in such a way that their combinations and mixtures are also contemplated and within the scope herein.

[00149] It should be understood that each maximum numerical limitation given throughout this descriptive report includes each lower numerical limitation, as if such lower numerical limitations were expressed Petition 870250084766, dated 09 / 19 / 2025, p. 45 / 149 37 / 115 clearly written here. Each minimum numerical limitation given throughout this descriptive report will include each higher numerical limitation, as if such higher numerical limitations were expressly written in this document. Each numerical range given throughout this descriptive report will include each narrower numerical range that lies within such a wider numerical range, as if such narrower numerical ranges were all expressly written in this document.

[00150] All lists of items, such as, for example, lists of ingredients, are intended 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.

[00151] Trade names for components including various ingredients used in the present invention may be referenced herein. The inventors in the present document do not intend to be limited by materials under any specific trade name. Equivalent materials (e.g., those obtained from a different source under a different name or reference number) to those referenced by the trade name may be substituted and used in the descriptions in the present document. CEACAM mRNA measurements, methods and uses

[00152] According to one of its objectives, the present invention relates to methods and uses for selecting an individual who needs the same for cancer treatment with a therapeutic agent that targets CEACAM5. A therapeutic agent that targets CEACAM5 may be an anti-CEACAM5 antibody conjugated with a cytotoxic agent.

[00153] The methods and uses of the invention may comprise a step of determining the amount of gene expression of CEACAM5 Petition 870250084766, dated 09 / 19 / 2025, pp. 46 / 149 38 / 115 (gene transcript or RNA) in a tumor sample.

[00154] The methods and uses of the invention may comprise a step of determining the amount of CEACAM5 gene expression, such as the CEACAM5 mRNA level in a tumor sample.

[00155] The amount of CEACAM5 gene expression can be expressed as a quantity relative to the total gene expression in the tumor sample and relative to the total length of expressed DNA or any other known method.

[00156] In some embodiments, the present invention relates to a method for selecting an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, wherein the method comprises at least the following steps: (i) determining a gene expression level value of CEACAM5 in an isolated tumor sample obtained from said individual, (ii) comparing said determined value with a reference value and (iii) selecting said individual for cancer treatment if the determined value is above the reference value.

[00157] In some embodiments, the present invention relates to a method for selecting an individual in need thereof for cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated with a cytotoxic agent, wherein the method comprises at least the steps of: (i) to determine a value of a gene expression level of CEACAM5 in an isolated sample of a tumor obtained from said individual Petition 870250084766, dated 09 / 19 / 2025, page 47 / 149 39 / 115 duo, (ii) compare said value determined in step (i) with a reference value of a CEACAM5 gene expression level, (iii) select said individual for a CEACAM5 immunohistochemical staining (IHC) test if the value determined in step (i) is above the reference value of step (ii), (iv) determine an intensity of a CEACAM5 protein expression level with said CEACAM5 immunohistochemical staining (IHC) test in an isolated tumor sample obtained from said individual, (v) compare said intensity determined in step (iv) with a reference intensity, e. (vi) select said individual for cancer treatment if the intensity determined in step (iv) is above the reference intensity.

[00158] The tumor samples from step (i) and step (iv) may be the same or different samples.

[00159] In some embodiments, the present invention relates to a method for diagnosing an individual in need of such diagnosis as eligible for cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, wherein the method comprises at least the following steps: (i) determine a gene expression level of CEACAM5 in an isolated tumor sample obtained from said individual, (ii) compare said determined value with a reference value, and (iii) select said individual as eligible for cancer treatment if the determined value is above the reference value. Petition 870250084766, dated 09 / 19 / 2025, pp. 48 / 149 40 / 115 reference.

[00160] In some embodiments, the present invention relates to a method for diagnosing an individual in need of such diagnosis as eligible for cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, wherein the method comprises at least the following steps: (i) determine a gene expression level value for CEACAM5 in an isolated tumor sample obtained from said individual, (ii) compare said value determined in step (i) with a reference value for a CEACAM5 gene expression level, (iii) select said individual for a CEACAM5 immunohistochemical staining (IHC) test if the value determined in step (i) is above the reference value of step (ii), (iv) determine the intensity of a CEACAM5 protein expression level with said CEACAM5 immunohistochemical staining (IHC) test in an isolated tumor sample obtained from said individual, (v) compare said intensity determined in step (iv) with a reference intensity, and. (vi) select said individual for cancer treatment if the intensity determined in step (iv) is above the reference intensity.

[00161] In some embodiments, the present invention relates to a method for selecting and treating an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, wherein the method comprises at least the steps of: Petition 870250084766, dated 09 / 19 / 2025, pp. 49 / 149 41 / 115 (i) determine a gene expression level of CEACAM5 in an isolated tumor sample obtained from said individual, (ii) compare said determined value with a reference value, (iii) select said individual for cancer treatment if the determined value is above the reference value, and (iv) administer to said selected individual an effective amount of said ADC.

[00162] In some embodiments, the present invention relates to a method for selecting and treating an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, wherein the method comprises at least the steps of: (i) determine a gene expression level value for CEACAM5 in an isolated tumor sample obtained from said individual, (ii) compare said value determined in step (i) with a reference value for a CEACAM5 gene expression level, (iii) select said individual for a CEACAM5 immunohistochemical staining (IHC) test if the value determined in step (i) is above the reference value of step (ii), (iv) determine the intensity of a CEACAM5 protein expression level with said CEACAM5 immunohistochemical staining (IHC) test in an isolated tumor sample obtained from said individual, (v) compare said intensity determined in step (iv) with a reference intensity, and. (vi) select the said individual for treatment of a Petition 870250084766, dated 09 / 19 / 2025, page 50 / 149 42 / 115 cancer if the intensity determined in step (iv) is above the reference intensity, and (vii) administer to said selected individual an effective amount of said ADC.

[00163] In some embodiments, the present invention relates to an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, for use in the treatment of cancer in an individual in need thereof, the use comprising: (i) to determine a gene expression level of CEACAM5 in an isolated tumor sample obtained from said individual, (ii) to compare said determined value with a reference value and (iii) to administer to said individual an effective amount of said ADC if said determined value is above the reference value.

[00164] In some embodiments, the present invention relates to an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, for use in the treatment of cancer in an individual in need thereof, the use comprising: (i) determine a value for a CEACAM5 gene expression level in an isolated tumor sample obtained from said individual, (ii) compare said value determined in step (i) with a reference value for a CEACAM5 gene expression level, (iii) select said individual for a CEACAM5 immunohistochemical staining (IHC) test if the value determined in step (i) is above the reference value of step (ii), Petition 870250084766, dated 09 / 19 / 2025, page 51 / 149 43 / 115 (iv) determine the intensity of a CEACAM5 protein expression level using said CEACAM5 immunohistochemical staining (IHC) test in an isolated tumor sample obtained from said individual, (v) compare said intensity determined in step (iv) with a reference intensity, e. (vi) administer to said individual an effective amount of said DCA if said determined intensity is above the reference intensity.

[00165] In some embodiments, the present invention relates to the use of a measurement of a value of the expression level of the CEACAM5 gene in an isolated tumor sample obtained from an individual in need thereof to characterize said tumor as a CEACAM5-high expressing tumor.

[00166] In some embodiments, the present invention relates to the use of a measurement of a value of the expression level of the CEACAM5 gene in an isolated tumor sample obtained from an individual in need thereof in order to select said individual for an immunohistochemical staining test for CEACAM5 (IHC).

[00167] In some embodiments, the present invention relates to the use of a measurement of a value of the expression level of the CEACAM5 gene in an isolated tumor sample obtained from an individual in need thereof in order to select said individual for cancer treatment with a CEACAM5-targeting therapeutic agent, for example, an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent.

[00168] In some embodiments, the present invention relates to the use of a measurement of a value of the expression level of the CEACAM5 gene in an isolated tumor sample obtained from an individual. Petition 870250084766, dated 09 / 19 / 2025, page 52 / 149 44 / 115 duo with the need for the same to select said individual for cancer treatment with a CEACAM5-targeting therapeutic agent, for example, an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent.

[00169] In some modalities, the value of the CEACAM5 gene expression level may be a measure of a CEACAM5 gene transcript.

[00170] In some embodiments, the CEACAM5 gene transcript may be an mRNA.

[00171] In some embodiments, the methods and uses of the invention comprise a step of determining a value of a CEACAM5 mRNA level in an isolated tumor sample.

[00172] In some embodiments, the amount of CEACAM5 gene expression can be expressed as a quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2.

[00173] The determined value or level can be compared to a limit value or reference value.

[00174] A deviation from the reference value may be indicative of a tumor that expresses CEACAM5 at a level sufficient to make it responsive to treatment that targets CEACAM5.

[00175] In the case of the CEACAM5 immunohistochemical staining test (IHC), the determined intensity or protein expression level can be compared to a threshold intensity or reference intensity.

[00176] A deviation from the reference intensity may be indicative of a tumor expressing CEACAM5 at a level sufficient to make it responsive to a treatment that targets CEACAM5-expressing cancer. Petition 870250084766, dated 09 / 19 / 2025, page 53 / 149 45 / 115

[00177] The methods and uses of the invention may be to characterize an individual in need thereof as being responsive to treatment targeting CEACAM5, for example, an anti-CEACAM5 antibody conjugated with a cytotoxic agent.

[00178] The methods and uses of the invention may include selecting an individual who needs the same as being responsive to a treatment targeting CEACAM5, for example, an anti-CEACAM5 antibody conjugated to a cytotoxic agent.

[00179] The methods and uses of the invention may be for monitoring the responsiveness of an individual in need of such to a treatment that targets CEACAM5, for example, an anti-CEACAM5 antibody conjugated with a cytotoxic agent.

[00180] The methods and uses of the invention may be to select a treatment that targets CEACAM5, for example an anti-CEACAM5 antibody conjugated with a cytotoxic agent, according to a level of CEACAM5 gene expression in a tumor of an individual who needs it.

[00181] The methods and uses of the invention may be to select a treatment that targets CEACAM5, for example an anti-CEACAM5 antibody conjugated with a cytotoxic agent, according to the intensity of CEACAM5 protein expression in a tumor of an individual who needs it.

[00182] The methods and uses of the invention may be to characterize a tumor in an individual who needs it as being responsive to a treatment targeting CEACAM5, for example, an anti-CEACAM5 antibody conjugated with a cytotoxic agent.

[00183] The methods and uses of the invention are performed on an isolated biological sample. An isolated sample can be an isolated sample from a tumor. A sample is isolated prior to the implementation of the methods and uses as disclosed in this document. Petition 870250084766, dated 09 / 19 / 2025, page 54 / 149 46 / 115

[00184] The methods and uses of the invention are carried out in vitro.

[00185] According to one of its objectives, the present invention relates to a method for selecting an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, wherein the method comprises at least the following steps: (i) determine a quantile-normalized, log2-transformed Transcripts Per Million Kilobases (TPM) value for CEACAM5 mRNA in a single tumor sample obtained from said individual, (ii) compare said value with a reference value, and (iii) select said individual for cancer treatment if the determined value is above the reference value.

[00186] According to one of its objectives, the present invention relates to a method for selecting an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, wherein the method comprises at least the following steps: (i) determine a quantile-normalized, log2-transformed Transcripts Per Million Kilobases (TPM) value for CEACAM5 mRNA in a single tumor sample obtained from said individual, (ii) compare said value determined in step (i) with a reference value, (iii) select said individual for a CEACAM5 immunohistochemistry (IHC) staining test if the value determined in step (i) is above the reference value from step (ii), (iv) determine an intensity of an expression level. Petition 870250084766, dated 09 / 19 / 2025, page 55 / 149 47 / 115 of the CEACAM5 protein with said CEACAM5 immunohistochemical staining (IHC) test in an isolated tumor sample obtained from said individual, (v) compare said intensity determined in step (iv) with a reference intensity, and. (vi) select said individual for cancer treatment if the determined intensity is above the reference intensity.

[00187] According to one of its objectives, the present invention relates to a method for diagnosing an individual in need of such diagnosis as being eligible for cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, wherein the method comprises at least the following steps: (i) determine a quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA in a single tumor sample obtained from said individual, (ii) compare said value with a reference value, and (iii) select said individual as eligible for cancer treatment if the determined value is above the reference value.

[00188] According to one of its objectives, the present invention relates to a method for diagnosing an individual in need of such diagnosis as being eligible for cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, wherein the method comprises at least the following steps: (i) determine a quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 to Petition 870250084766, dated 09 / 19 / 2025, page 56 / 149 48 / 115 mRNA of CEACAM5 in an isolated tumor sample obtained from said individual, (ii) compare said value determined in step (i) with a reference value, (iii) select said individual for a CEACAM5 immunohistochemical staining (IHC) test if the value determined in step (i) is above the reference value of step (ii), (iv) determine an intensity of expression level of the CEACAM5 protein with said CEACAM5 immunohistochemical staining (IHC) test in an isolated tumor sample obtained from said individual, (v) compare said intensity determined in step (iv) with a reference intensity, and. (vi) select said individual for cancer treatment if the determined intensity is above the reference intensity.

[00189] According to another of its objectives, the present invention relates to a method for selecting and treating an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated with a cytotoxic agent, wherein the method comprises at least the steps of: (i) determine a quantile-normalized, log2-transformed Transcripts Per Million Kilobases (TPM) value for CEACAM5 mRNA in a single tumor sample obtained from said individual, (ii) compare said value with a reference value, (iii) select said individual for cancer treatment if the determined value is above the reference value, and (iv) administer to said selected individual a quantity Petition 870250084766, dated 09 / 19 / 2025, page 57 / 149 49 / 115 of the effectiveness of said ADC.

[00190] According to another of its objectives, the present invention relates to a method for selecting and treating an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated with a cytotoxic agent, wherein the method comprises at least the steps of: (i) determine a quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA in a single tumor sample obtained from said individual, (ii) compare said value determined in step (i) with a reference value, (iii) select said individual for a CEACAM5 immunohistochemical staining (IHC) test if the value determined in step (i) is above the reference value of step (ii), (iv) determine an intensity of a CEACAM5 protein expression level with said CEACAM5 immunohistochemical staining (IHC) test in a single tumor sample obtained from said individual, (v) compare said intensity determined in step (iv) with a reference intensity, and. (vi) select said individual for cancer treatment if the determined intensity is above the reference intensity, and (vii) administer to said selected individual an effective amount of said ADC.

[00191] In accordance with another of its objectives, the present invention relates to an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, Petition 870250084766, dated 09 / 19 / 2025, page 58 / 149 50 / 115 for use in the treatment of cancer in an individual in need thereof, the use comprises (i) determining a quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA in an isolated tumor sample obtained from said individual, (ii) comparing said value with a reference value, and (iii) administering to said individual an effective amount of said ADC if the determined value is above the reference value.

[00192] In accordance with another of its objectives, the present invention relates to an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, for use in the treatment of cancer in an individual in need thereof, the use comprising: (i) determine a quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA in a single tumor sample obtained from said individual, (ii) compare said value determined in step (i) with a reference value, (iii) select said individual for a CEACAM5 immunohistochemical staining (IHC) test if the value determined in step (i) is above the reference value of step (ii), (iv) determine an intensity of a CEACAM5 protein expression level with said CEACAM5 immunohistochemical staining (IHC) test in a single tumor sample obtained from said individual, (v) compare said intensity determined in step (iv) with a reference intensity, and. Petition 870250084766, dated 09 / 19 / 2025, page 59 / 149 51 / 115 (vi) administer to said individual an effective amount of said DCA if the determined intensity is above the reference intensity.

[00193] In some modalities, the reference value may be at least about 7 to about 13.

[00194] In some embodiments, the preference value may be at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, at least about 12 or at least about 13.

[00195] In some embodiments, quantile normalization can be obtained by (i) classifying sample transcripts by expression level, (ii) calculating an average value for genes occupying the same class, and (iii) replacing the values ​​of all genes occupying the same class with that average value.

[00196] In some modalities, the expression level of transcripts can be measured in Fragments per Million Kilobases (FPKM) before being converted to TPM.

[00197] In some embodiments, Fragments per Million Kilobases (FPKM) can be obtained by counting the total number of transcripts in the sample, dividing the transcript counts obtained by 1,000,000, and dividing the values ​​obtained by the gene length, in kilobases.

[00198] One step of the methods and uses disclosed in this document is to determine the relative expression of the CEACAM5 gene in relation to other genes in a tumor sample. Many methods can be used for such a determination.

[00199] The relative amount of CEACAM5 gene transcript is then compared to a reference value. A significant deviation from the reference value may be indicative of a CEACAM5-expressing tumor that may be responsive to a therapeutic agent that has Petition 870250084766, dated 09 / 19 / 2025, page 60 / 149 52 / 115 specifically targeting CEACAM5.

[00200] Gene transcript quantities can be measured by any methods known in the art, such as microarrays, large-scale real-time reverse transcription PCR, RNA sequencing (RNA-Seq), next-generation sequencing (NGS)

[00201] The gene expression (RNA-seq) of a tumor sample can be obtained by any methods known in the art.

[00202] RNA-Seq is a sequencing method used to determine gene expression levels. The number of reads determined to originate from each transcript (usually by alignment) is proportional to its expression level. RNA-Seq can be used to generate a gene expression profile for tumor samples in many types of cancer and to determine which gene expression levels are responsible for tumor development. RNA-Seq data can be harmonized by aligning raw RNA reads to the GRCh38 reference genome construct and calculating gene expression levels with standardized protocols. RNA-Seq data may be available as aligned reads (BAM) and expression levels as: raw and normalized counts with TPM, FPKM, or FPKM-UQ.

[00203] In some modalities, gene expression (RNA-seq) can be obtained as follows.

[00204] Gene transcripts (RNA) from an isolated tumor sample can be sequenced using the KAPA HYPERPREPKITILLUMINA® mRNA Platform. RNA-seq data can be processed as follows: sequencing reads can be mapped to the GRCh 38 reference genome using, for example, Reference Alignment of Spliced ​​Transcripts (STAR)

[25] . Gene expression can be first Petition 870250084766, dated 09 / 19 / 2025, page 61 / 149 53 / 115 measured in FPKM (Fragments per Million Kilobases) by CUFFLINK

[26] and gene-level FPKM can be converted to TPM (Transcripts per Million Kilobases)

[27] . TPM values ​​can be log2 transformed and quantile normalized for downstream analysis, including differential gene expression (DGE) analysis. In some modalities, samples with a number of genes detected below 10000 can be excluded from downstream analysis. RNA-seq can include counter-analysis of microenvironment cell populations [MCP], according to published methods [28, 29].

[00205] In some modalities, the measurement of the CEACAM5 gene expression level (mRNA or gene transcription level measurement) can be normalized.

[00206] In some modalities, the measurement of the gene expression level of CEACAM5 (mRNA or measurement of the gene transcription level) can be transformed by log2.

[00207] In some modalities, the expression level of the CEACAM5 gene (mRNA) in a tumor sample is expressed as a quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2

[00208] Normalization of the CEACAM5 gene expression level measurement can be achieved with the following steps: - Log2 transformation of raw RNA-seq count values, - Quantitative normalization of RNA-seq data by matching the distribution of expression values ​​between samples by classifying the values ​​and adjusting them to have the same distribution, and - Expression of data as Transcripts Per Million Kilobases (TPM) by scaling the expression values ​​normalized by gene length and the total number of sequential reads. Petition 870250084766, dated 09 / 19 / 2025, page 62 / 149 54 / 115 adas and then scaling to one million to make the values ​​more interpretable.

[00209] In some modalities, the quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for the CEACAM5 gene expression level measured (or determined) in a tumor is compared to a reference value. If the determined value is above the reference value, the tumor may qualify as being responsive to CEACAM5-targeted treatment. An individual who needs it and whose determined value is above the reference value may be selected for CEACAM5-targeted treatment.

[00210] In some modalities, the reference value may be at least about 7 to about 13.

[00211] In some embodiments, the preference value may be at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, at least about 12 or at least about 13.

[00212] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA may be approximately =7.

[00213] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA may be approximately =7.5.

[00214] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA may be approximately =8.

[00215] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA may be approximately =8.5.

[00216] In some modalities, the value of Transcribed By Me Petition 870250084766, dated 09 / 19 / 2025, p. 63 / 149 55 / 115 million kilobases (TPM) normalized by quantile transformed by log2 for CEACAM5 mRNA may be approximately =9.

[00217] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA may be approximately =9.5.

[00218] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA may be approximately =10.

[00219] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA may be approximately =10.5.

[00220] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA may be approximately =11.

[00221] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA may be approximately =11.5.

[00222] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA may be approximately =12.

[00223] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA may be approximately =12.5

[00224] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA may be approximately =13.

[00225] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA may be approximately =13.5.

[00226] In some modalities, the value of Transcribed By Me Petition 870250084766, dated 09 / 19 / 2025, p. 64 / 149 56 / 115 million kilobases (TPM) normalized by quantile transformed by log2 for CEACAM5 mRNA may be approximately =14.

[00227] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA may be approximately =14.5.

[00228] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA may be approximately =15.

[00229] In some embodiments, the quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA can be around >7.

[00230] In some embodiments, the quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA can be around >7.5.

[00231] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA can be around >8.

[00232] In some embodiments, the quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA can be around >8.5.

[00233] In some embodiments, the quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA can be around >9.

[00234] In some embodiments, the quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA can be around >9.5.

[00235] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA can be around >10.

[00236] In some modalities, the value of Transcribed By Me Petition 870250084766, dated 09 / 19 / 2025, p. 65 / 149 57 / 115 million kilobases (TPM) normalized by quantile transformed by log2 for CEACAM5 mRNA may be approximately >10.5.

[00237] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA can be around >11.

[00238] In some embodiments, the quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA can be around >11.5.

[00239] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA can be around >12.

[00240] In some embodiments, the value of quantile-normalized Transcripts Per Million Kilobases (TPM) transformed by log2 for CEACAM5 mRNA can be approximately >12.5

[00241] In some embodiments, the quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA can be around >13.

[00242] In some embodiments, the quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA can be around >13.5.

[00243] In some embodiments, the quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA can be around >14.

[00244] In some embodiments, the quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA can be around >14.5.

[00245] In some embodiments, the quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA can be around >15.

[00246] The level (or intensity) of CEA protein expression Petition 870250084766, dated 09 / 19 / 2025, p. 66 / 149 58 / 115 CAM5 can be measured by immunohistochemistry (IHC).

[00247] The level and pattern of CEACAM5 protein expression in an isolated tumor sample from a patient in need of the same can be analyzed by immunohistochemistry (IHC), for example, as revealed by LaPointe et al. (Journal of Clinical Oncology, Volume 39, number 15_suppl., https: / / doi.org / 10.1200 / JCQ.2021.39.15_suppl.e21030) or by Blumenthal et al. (BMC Cancer. 2007;7:2. Published January 3, 2007. doi:10.1186 / 1471-2407-7-2).

[00248] CEACAM5 reactivity can be assessed in tumor cells using semi-quantitative Percent Scores (calculated by summing the percentages of intensities >2+) or H-score for CEACAM5 plasma membrane staining (whole or polarized).

[00249] High CEACAM5 expressers can be defined as patients with CEACAM5 protein expression > 2+ in > 50% of the tumor cell population of a tumor sample.

[00250] Moderate CEACAM5 expressers can be defined as patients with CEACAM5 protein expression > 2+ in > 1% to < 50% of the tumor cell population in a tumor sample.

[00251] In some modalities, a reference intensity of a CEACAM5 protein expression level measured with a CEACAM5 immunohistochemical staining (IHC) test may be an intensity of > 2+ in > 50% of the tumor cell population.

[00252] A patient requiring cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent that has CEACAM5 expression at an intensity of >2+ in >50% of the tumor cell population may be selected for said treatment. Petition 870250084766, dated 09 / 19 / 2025, page 67 / 149 59 / 115 treatment.

[00253] A patient requiring cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent that has CEACAM5 expression at an intensity of > 2+ in > 50% of the tumor cell population may be administered for said treatment.

[00254] A patient requiring the same for cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated with a cytotoxic agent may first undergo a determination of CEACAM5 gene expression level in an isolated tumor sample obtained from said patient. If the determined value of CEACAM5 gene expression level in an isolated tumor sample obtained from said patient is above a reference value, the patient may be selected for a subsequent test to determine the intensity of CEACAM5 protein expression level with a CEACAM5 immunohistochemical staining (IHC) test.

[00255] If the determined intensity is above a reference intensity, the patient may be selected for treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated with a cytotoxic agent.

[00256] If the determined intensity is above a reference intensity, the patient may be administered treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated with a cytotoxic agent. Antibody-drug conjugate comprising an anti-CEACAM5 antibody

[00257] The present invention relates to an anticor combination. Petition 870250084766, dated 09 / 19 / 2025, pp. 68 / 149 60 / 115 po-drug (ADC) comprising an anti-CEACAM5 antibody or a fragment thereof.

[00258] 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 antibody-drug conjugate, the anti-CEACAM5 antibody may be covalently linked via a cleavable or non-cleavable ligand to at least one chemotherapeutic agent. Anti-CEACAM5 antibody

[00259] According to one embodiment, the antibody-drug conjugate comprises an anti-CEACAM5 antibody or a fragment thereof.

[00260] According to one embodiment, the antibody-drug conjugate comprises a humanized anti-CEACAM5 antibody or a fragment thereof.

[00261] According to one embodiment, the antiCEACAM5 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 the amino acid sequence NTR and CDR-L3 consisting of SEQ ID NO: 5.

[00262] In a further embodiment, the anti-CEACAM5 antibody or a fragment thereof comprises a variable heavy chain (VH) domain that has at least 90% identity with SEQ ID NO: 6 and a variable light chain (VL) domain that has at least 90% identity with SEQ ID NO: 7, wherein CDR1-H consists of SEQ ID NO: 1, CDR2-H consists of SEQ ID NO: 2, CDR3H consists of SEQ ID NO: 3, CDR1-L consists of SEQ ID NO: 4, CDR2-L consists of the amino acid sequence NTR and CDR3-L consists Petition 870250084766, dated 09 / 19 / 2025, p. 69 / 149 61 / 115 te na SEQ ID NO: 5.

[00263] In a further embodiment, the anti-CEACAM5 antibody or a fragment thereof comprises a variable heavy chain (VH) domain having at least about 92%, at least 95%, at least 98% identity with SEQ ID NO: 6 and a variable light chain (VL) domain having at least 92%, at least 95%, at least 98% identity with 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 the amino acid sequence NTR and CDR3-L consists of SEQ ID NO: 5.

[00264] In a further embodiment, the anti-CEACAM5 antibody or a fragment thereof comprises a variable heavy chain (VH) domain consisting of SEQ ID NO: 6 and a variable light chain (VL) domain consisting of SEQ ID NO: 7.

[00265] The anti-CEACAM5 antibody or a fragment thereof comprises in a further embodiment: - a variable heavy chain domain consisting of sequence EVQLQESGPGLVKPGGSLSLSCAASGFVFSSYDMSWVRQTPERGL EWVAYISSGGGITYAPSTVKGRFTVSRDNAKNTLYLQMNSLTSEDTA VYYCAAHYFGSSGPFAYWGQGTLVTVSS (SEQ ID NO: 6, with CDRs shown in bold characters) wherein 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 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, CDR3H spans amino acid positions 97 to 109 (SEQ ID NO: 3), and FR4-H spans amino acid positions 110 to 120, and - a variable light chain domain consisting of se Petition 870250084766, dated 09 / 19 / 2025, p. 70 / 149 62 / 115 frequency DIQMTQSPASLSASVGDRVTITCRASENIFSYLAWYQQKPGKSPKLL VYNTRTLAEGVPSFSGSGSGTDFSLTISSLQPEDFATYYCQHHYGTP FTFGSGTKLEIK (SEQ ID NO: 7, with CDRs shown in bold characters) where FR1-L covers amino acid positions 1 to 26, CDR1-L covers amino acid positions 27 to 32 (SEQ ID NO: 4), FR2-L covers amino acid positions 33 to 49, CDR2-L covers amino acid positions 50 to 52, FR3-L covers amino acid positions 53 to 88, CDR3-L covers amino acid positions 89 to 97 (SEQ ID NO: 5) and FR4-L covers amino acid positions 98 to 107.

[00266] In a further embodiment, the anti-CEACAM5 antibody or fragment thereof comprises a heavy chain (HC) having at least 90% sequence identity with SEQ ID NO: 8 and a light chain (LC) having at least 90% sequence identity with SEQ ID NO: 9, wherein 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 the amino acid sequence NTR and CDR3-L consists of SEQ ID NO: 5.

[00267] In a further embodiment, the anti-CEACAM5 antibody or a fragment thereof comprises a heavy chain (HC) having at least about 92%, at least 95%, at least 98% identity with SEQ ID NO: 8 and a light chain (LC) having at least 92%, at least 95%, at least 98% identity with 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, CDR1L consists of SEQ ID NO: 4, CDR2-L consists of the amino acid sequence NTR and CDR3-L consists of SEQ ID NO: 5.

[00268] In a further embodiment, the anti-CEACAM5 antibody or a fragment thereof comprises a heavy chain (HC) Petition 870250084766, dated 09 / 19 / 2025, p. 71 / 149 63 / 115 consisting of SEQ ID NO: 8 and a light chain (LC) consisting of SEQ ID NO: 9.

[00269] 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) comprising the CDR1-H, CDR2-H and CDR3H of the antibodies as described above. The antibody may also be a heavy-chain antibody, i.e., an antibody devoid of a light chain that may or may not contain a CH1 domain.

[00270] 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.

[00271] 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 diacorpos.

[00272] The antibody may also be a bispecific or multispecific antibody formed from antibody fragments, wherein at least one antibody fragment is an antibody fragment according to the invention. Multispecific antibodies are polyvalent protein complexes as described, for example, in document EP 2 050 764 A1 or US 2005 / 0003403 A1.

[00273] Anti-CEACAM5 antibodies and fragments thereof can be produced by any technique well known in the art. In particular, said antibodies are produced by techniques as described hereinafter in this document.

[00274] The anti-CEACAM5 antibody and fragments thereof can be used in an isolate (e.g., purified) of or contained in a vector, such as a membrane or lipid vesicle (e.g., a Petition 870250084766, dated 09 / 19 / 2025, page 72 / 149 64 / 115 liposome).

[00275] 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 alone or in combination.

[00276] Knowing the amino acid sequence of the desired sequence, a person skilled in the art can readily produce the anti-CEACAM5 antibody and fragments thereof by standard polypeptide production techniques. For example, they can be synthesized using a well-known solid-phase method, in particular, using a commercially available peptide synthesis apparatus (such as one manufactured by Applied Biosystems, Foster City, California) and following the manufacturer's instructions. Alternatively, the anti-CEACAM5 antibody and fragments thereof can be synthesized by recombinant DNA techniques as known in the art.For example, these fragments can be obtained as DNA expression products after incorporating DNA sequences encoding the desired (poly)peptide into expression vectors and introducing such vectors into suitable eukaryotic or prokaryotic hosts that will express the desired polypeptide from which they can be isolated using well-known techniques.

[00277] Anti-CEACAM5 antibody and fragments thereof are appropriately 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.

[00278] Methods for producing humanized antibodies based on conventional recombinant DNA are well known in the art (See, for example, Riechmann L. et al. 1988; Neuberger MS. et al. 1985). Antibodies can be humanized using a variety of Petition 870250084766, dated 09 / 19 / 2025, page 73 / 149 65 / 115 of techniques known in the art including, for example, the technique disclosed in application WO2009 / 032661, CDR grafting (document EP 239,400; PCT publication WO91 / 09967; US Patent Nos. 5,225,539; 5,530,101; and 5,585,089), surface modification or recoating (EP 592,106; EP 519,596; Padlan EA (1991); Studnicka GM et al. (1994); Roguska MA. et al. (1994)), and chain scrambling (Pat. No. US 5,565,332). The general recombinant DNA technology for preparing such antibodies is also known (see European Patent Application EP 125023 and International Patent Application WO 96 / 02576).

[00279] The anti-CEACAM5 antibody Fab can be obtained by treating an antibody that reacts specifically with CEACAM5 with a protease, such as papain. Furthermore, the anti-CEACAM5 antibody Fab can be produced by inserting DNA sequences encoding both strands of the anti-CEACAM5 antibody Fab into a vector for prokaryotic or eukaryotic expression, and introducing the vector into prokaryotic or eukaryotic cells (as appropriate) to express the anti-CEACAM5 antibody Fab.

[00280] The OF(ab')2 of the anti-CEACAM5 antibody can be obtained by treating the antibody that reacts specifically with CEACAM5 with a protease, pepsin. Furthermore, the F(ab')2 of the anti-CEACAM5 antibody can be produced by linking Fab' as described below via a thioether linkage or a disulfide linkage.

[00281] The Fab' of the anti-CEACAM5 antibody can be obtained by treating F(ab')2, which reacts specifically with CEACAM5, with a reducing agent such as dithiothreitol. Furthermore, the Fab' of the anti-CEACAM5 antibody can be produced by inserting DNA sequences encoding antibody Fab chains into a vector for prokaryotic expression, or a vector for eukaryotic expression, and introducing the Petition 870250084766, dated 09 / 19 / 2025, page 74 / 149 66 / 115 vector in prokaryotic or eukaryotic cells (as appropriate) to carry out its expression.

[00282] The anti-CEACAM5 antibody scFv can be produced by sequencing 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 can be used, which involves selecting the complementarity-determining regions (CDRs) according to the invention and grafting them onto a human scFv fragment framework of known three-dimensional structure (see, for example, documents W098 / 45322; WO 87 / 02671; US5,859,205; US5,585,089; US4,816,567; EP0173494).

[00283] In one embodiment, the anti-CEACAM5 antibody is tusamitamab (CAS [2349294-95-5]. Chemotherapeutic agents

[00284] The antibody-drug conjugate for use according to the present invention typically comprises at least one chemotherapeutic agent (also referred to as a cytotoxic agent in this document). A chemotherapeutic agent as used in this document refers to an agent that kills cells, including cancer cells. Such agents favorably inhibit the division and growth of cancer cells and cause tumors to shrink in size. The term chemotherapeutic agent is used in this document interchangeably with the terms cytotoxic agent, growth inhibitor, or cytostatic drug.

[00285] The term chemotherapeutic agent as used in Petition 870250084766, dated 09 / 19 / 2025, page 75 / 149 67 / 115 This document refers to a substance that inhibits or prevents cell function and / or causes cell destruction. 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.

[00286] In an additional embodiment, the chemotherapeutic agent is selected from the group consisting of radioisotopes, protein toxins, small molecule toxins, and combinations thereof.

[00287] Radioisotopes include radioactive isotopes suitable for treating cancer. Such radioisotopes generally emit primarily beta radiation. In an additional embodiment, the radioisotopes are selected from the group consisting of At211, Bi212, Er169, I131, I125, Y90, In111, P32, Re186, Re188, Sm153, Sr89, radioactive isotopes of Lu and combinations thereof. In one embodiment, the radioactive isotope is an alpha-emitting isotope, more specifically, Th227, which emits alpha radiation.

[00288] In a further embodiment, small molecule toxins are selected from among antimetabolites, DNA alkylating agents, DNA crosslinking agents, DNA intercalating agents, antimicrotubular agents, topoisomerase inhibitors and combinations thereof.

[00289] In a further embodiment, the antimicrotubular agent is selected from the group consisting of taxanes, vinca alkaloids, maytansinoids, colchicine, podophyllotoxin, gruseofulvin and combinations thereof.

[00290] In some modalities, a chemotherapeutic agent may be a maytansinoid. Petition 870250084766, dated 09 / 19 / 2025, page 76 / 149 68 / 115

[00291] According to one embodiment, maytansinoids are selected from maytansinol, maytansinol analogues and combinations thereof.

[00292] 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. Patents 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.

[00293] In a further embodiment, the cytotoxic conjugates of the present invention utilize the thiol-containing maytansinoid (DM1), formally called N2'-deacetyl-N2'-(3-mercapto-1-oxopropyl)maytansine, as the cytotoxic agent. DM1 is represented by the following structural formula (I):

[00294] In a further embodiment, the cytotoxic conjugates of the present invention utilize the thiol-containing maytansinoid, DM4, formerly called N2'-deacetyl-N-2'(4-methyl-4-mercapto-1-oxopentyl)-maytansine, as the cytotoxic agent. DM4 is represented by the following structural formula (II): Petition 870250084766, dated 09 / 19 / 2025, page 77 / 149 69 / 115 T N I OH H M eO

[00295] In further embodiments of the invention, other maytansines may be used, including thiol- and disulfide-containing maytansinoids that carry a mono- or dialkyl substitution on the carbon atom that carries the sulfur atom. These include a maytansinoid that has, at C-3, C-14 hydroxymethyl, C-15 hydroxy or C-20 desmethyl, an acylated amino acid side chain with an acyl group containing a hindered sulfhydryl group, wherein the carbon atom of the acyl group containing the thiol functionality has one or two substituents, said substituents being linear or branched CH3, C2H5, alkyl or alkenyl having from 1 to 10 reactants and any aggregate that may be present in solution.

[00296] Examples of these cytotoxic agents and conjugation methods are further given in application WO 2008 / 010101 which is incorporated by reference.

[00297] The immunoconjugates according to the present invention can be prepared as described in application WO 2004 / 091668, the entire content of which is incorporated by reference herein.

[00298] Consequently, in a further embodiment, maytansinoids are selected from the group consisting of N2'-deacetyl- / V2'-(3-mercapto-1-oxopropyl)-maytansine (DM1) or N2'-deacetyl- / V-2'-(4-methyl-4-mercapto-1-oxopentyl)-maytansine (DM4) and Petition 870250084766, dated 09 / 19 / 2025, p. 78 / 149 70 / 115 combinations of them.

[00299] In an additional embodiment, in the antibody-drug conjugate, the anti-CEACAM5 antibody is covalently linked via a cleavable or non-cleavable ligand to at least one chemotherapeutic agent.

[00300] In a further embodiment, the ligand is selected from the group consisting of N-succinimidyl pyridyldithiobutyrate (SPDB), 4-(pyridine-2-yldisulfanyl)-2-sulfobutyric acid (sulfo-SPDB), and succinimidyl(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC).

[00301] In a further embodiment, the ligand binds to a lysine or cysteine ​​residue in the Fc region of the anti-CEACAM5 antibody. In a further embodiment, the ligand forms a disulfide bond or a thioether bond with maytansine.

[00302] In particular, the anti-CEACAM5drug antibody conjugate may be selected from the group consisting of: the anti-CEACAM5-SPDB-DM4 antibody-drug conjugate of formula (III): anti-CEACAM5-SPDB-DM4 J anti-CEACAM5-sulfo-SPDB-DM4-drug antibody conjugate of formula (IV): Petition 870250084766, dated 09 / 19 / 2025, p. 79 / 149 71 / 115 anti-CEACAM5-sulfo-SPDBDM4(|V); and an anti-CEACAM5-SMCC-DM1-drug antibody conjugate of formula (V): anti-C EACAM5-SMCC(V).

[00303] In formulas (III), (IV) and (V) above, n corresponds to the number of chemotherapeutic agent molecules conjugated per antibody molecule. It corresponds to the drug-to-antibody ratio (or DAR) defined below and can vary from 1 to 10.

[00304] In a further embodiment, the antibody-drug conjugate of the present invention comprises an anti-CEACAM5 antibody, comprising a heavy chain (VH) of SEQ ID NO: 8 and a light chain (VL) of SEQ ID NO: 9 (tusamitamab), wherein Petition 870250084766, dated 09 / 19 / 2025, p. 80 / 149 72 / 115 tusamitamab is covalently linked to N2'-deacetyl-N-2'(4-methyl-4-mercapto-1-oxopentyl)-maitansine (DM4) via succinimidyl pyridyldibutyrate (SPDB). In this way, the antibody-drug conjugate, tusamitamab ravtansine, (huMAb2-3-SPDB-DM4) is obtained.

[00305] In one embodiment, the antibody-drug conjugate of the present invention is tusamitamab ravtansine (CAS [2254086-60-5]).

[00306] Ligand, as used in this document, means a chemical moiety comprising a covalent bond or a chain of atoms that covalently links the antibody to the chemotherapeutic agent moiety (e.g., a cytostatic agent, a cytotoxic agent, or a growth-inhibiting agent). Suitable ligands are well known in the art and include disulfide groups, thioether groups, acid-labile groups, photolabile groups, peptidase-labile groups, and esterase-labile groups.

[00307] Conjugates can be prepared by in vitro methods. In order to link a drug or prodrug to an antibody, for example, a chemotherapeutic agent, a linking group is used. Suitable linking groups well known in the art include disulfide groups, thioether groups, acid-labile groups, photolabile groups, peptidase-labile groups, and esterase-labile groups.The conjugation of an antibody with a chemotherapeutic agent of the invention, such as a cytotoxic agent, can be done 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 (nitroSPDB), 4-(pyridin-2-yldisulfanyl)-2-sulfobutyric acid (sulfo-SPDB), N-succinimidyl (2-pyridyldithio)propionate (SPDP), succinimidyl (N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC), iminothiolane (IT), bifunctional imidoester derivatives (such as dimethyl adipimidate HCl), active esters (such as disuccinimidyl suberate), village. Petition 870250084766, dated 09 / 19 / 2025, page 81 / 149 73 / 115 of (such as glutaraldehyde), bis-azido compounds (such as bis(p-azidobenzoyl)-hexanediamine), bis-diazonium derivatives (such as bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as 2,6-diisocyanate toluene) and bisactive 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-isothiocyanatobenzylmethyldiethylenetriaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for radionucleotide-to-antibody conjugation (WO 94 / 11026).

[00308] The ligand may be a cleavable ligand that facilitates the release of the chemotherapeutic agent into the cell. For example, an acid-labile ligand, a peptidase-sensitive ligand, an esterase-labile ligand, a photolabile ligand, or a disulfide-containing ligand may be used (see, for example, U.S. Patent No. 5,208,020). The ligand may also be a non-cleavable ligand (e.g., SMCC ligand) which may lead to better tolerance in some cases.

[00309] In general, the conjugate can be obtained by a process that includes the following steps: (i) placing an optionally buffered aqueous solution of a cell-binding agent (e.g., an antibody according to the invention) in contact with solutions of a ligand and a chemotherapeutic agent, such as a cytotoxic compound (or agent); (ii) then optionally separate the conjugate that was formed (i) from the unreacted cell-binding agent (e.g., antibody of the invention) and unreacted chemotherapeutic agent, as unreacted cytotoxic compound (or agent).

[00310] The aqueous solution of cell-binding agent can be buffered with buffers such as, for example, phosphate, acetate, potassium citrate or N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (Hepes buffer). The buffer depends on the nature of the Petition 870250084766, dated 09 / 19 / 2025, page 82 / 149 74 / 115 cell-binding agent (e.g., antibody of the invention). The chemotherapeutic agent, such as the cytotoxic compound (or agent), is in solution in a polar organic solvent, for example, dimethyl sulfoxide (DMSO) or dimethylacetamide (DMA).

[00311] The reaction temperature is 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.

[00312] Numerous different chromatography methods can be used by a person skilled in the art in order to perform the separation of step (ii): the conjugate can be purified, for example, from aggregates, for example, 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.

[00313] As used in this document, the term aggregate means associations that can be formed by two or more cell-binding agents, wherein said agents are modified or not by conjugation. Aggregates can be formed under the influence of a large number of parameters, such as a high concentration of cell-binding agent (e.g., antibody of the invention) in the solution, the pH of the solution, high shear forces, the number of linked dimers and their hydrophobic character, the temperature (see Wang & Gosh, 2008, J. Membrane Sci., 318: 311-316 and references cited therein); it is noted that the relative influence of Petition 870250084766, dated 09 / 19 / 2025, page 83 / 149 75 / 115 some of these parameters are 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 person skilled, such as SEC (see Walter et al., 1993, Anal. Biochem., 212(2): 469-480).

[00314] After step (i) or (ii), the solution containing the conjugate may be subjected to an additional step (iii) of chromatography, ultrafiltration and / or diafiltration.

[00315] The conjugate is recovered at the end of these steps in an aqueous solution.

[00316] In a further embodiment, the antibody-drug conjugate according to the invention 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 for conjugates including maytansinoid molecules.

[00317] This DAR number can vary with the nature of the antibody and the drug (i.e., the chemotherapeutic agent, such as a cytotoxic agent or a growth inhibitor) used together with the experimental conditions used for conjugation (such as the ratio of chemotherapeutic agent (e.g., growth inhibitor) / antibody, the reaction time, the nature of the solvent and cosolvent, if any). Thus, the contact between the antibody and the chemotherapeutic agent, such as a cytotoxic agent or a growth inhibitor, leads to a mixture comprising several conjugates that differ from each other by different drug-to-antibody ratios; optionally, the naked antibody; optionally aggregated. The DAR that is determined is thus an average value.

[00318] One method that can be used to determine the DAR consists of spectrophotometrically measuring the absorbance ratio. Petition 870250084766, dated 09 / 19 / 2025, page 84 / 149 76 / 115 of a substantially purified conjugate solution at ΔD and 280 nm. 280 nm is a wavelength generally used to measure protein concentration, such as antibody concentration. The wavelength ΔD is selected in order to allow discrimination of the drug from the antibody, that is, as readily known to one skilled in the art, ΔD is a wavelength at which the drug (i.e., chemotherapeutic agent) has a high absorbance and ΔD is sufficiently far from 280 nm to avoid substantial overlap in the absorbance peaks of the drug and antibody. ΔD may be selected as 252 nm in the case of maytansinoid molecules. A method for calculating ΔD can be derived from Antony S. Dimitrov (ed), LLC, 2009, Therapeutic Antibodies and Protocols, volume 525, 445, Springer Science:

[00319] The absorbances for the conjugate at AD (AAD) and at 280 nm (A280) are measured at the monomeric peak of the size exclusion chromatography (SEC) analysis (allowing calculation of the DAR(SEC) parameter) or using a classical spectrophotometer instrument (allowing calculation of the DAR(UV) parameter). The absorbances can be expressed as follows: AÀD = (cD x εDÀD) + (cA x εAÀD) A280 = (cD x εD280) + (cA x εA280) where: cD and cA are respectively the concentrations in the solution of the drug (i.e., chemotherapeutic agent) and the antibody; εDÀD and εD280 are, respectively, the molar extinction coefficients of the drug at ÀD and 280 nm; εAÀD and εA280 are, respectively, the molar extinction coefficients of the antibody at ÀD and 280 nm.

[00320] Solving these two equations with two unknowns leads to the following equations: Petition 870250084766, dated 09 / 19 / 2025, p. 85 / 149 77 / 115 cD = [(εA280 x AAD) - (εAλD x A280)] / [(εDλD x εΑ280) - (εAλD x εD280)] cA = [A280 - (cD x εD280)] / εΑ280

[00321] The average DAR is then calculated from the ratio of the drug concentration to that of the antibody: DAR = cD / cA.

[00322] 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.

[00323] 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.

[00324] In some embodiments, the dosing regimen may comprise administering the dose over a period of about 10 minutes to about 48 hours or from about 1 hour to about 48 hours, as in a period of 1 hour to 4 hours. In some embodiments, the dosing regimen may comprise administering a dose over a period of about 1 hour.

[00325] In some embodiments, the antibody-drug conjugate comprising an anti-CEACAM5 antibody may be administered over a period of time ranging from about 30 minutes to about 3 hours, or from about 1 hour to about 2 hours, or for about 1.5 hours. ADC dosages

[00326] In some embodiments, an antibody-drug conjugate is disclosed comprising an anti-CEACAM5 antibody and a chemotherapeutic agent for use in treating cancer in combination with an anti-CTLA4 antibody and, when present, an anti-PD-1- or anti-PD-L1 antibody, wherein the antibody-drug conjugate may be Petition 870250084766, dated 09 / 19 / 2025, p. 86 / 149 78 / 115 administered at a dose of 60 mg / m2 to 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 ​​an individual who needs it.

[00327] In some embodiments, the antibody-drug conjugate may be administered at a dose of approximately 60 to approximately 210 mg / m2 or approximately 80 to approximately 170 mg / m2 or approximately 100 to approximately 150 mg / m2.

[00328] In several embodiments, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 60, 70, 80, 90, 100, 110, 120, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205 or approximately 210 mg / m2.

[00329] In several embodiments, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 60, 80, 100, 120, 135, 150, 170, 180, 190 or approximately 210 mg / m2.

[00330] In some embodiments, ADC may be administered at a dose of approximately 80 mg / m2 to approximately 170 mg / m2, or at a dose of approximately 80 mg / m2 to approximately 150 mg / m2, or at a dose of approximately 100 mg / m2 to approximately 120 mg / m2.

[00331] In some embodiments, ADC may be administered at a dose of approximately 80 mg / m2 or approximately 100 mg / m2 or approximately 120 mg / m2 or approximately 150 mg / m2 or approximately 170 mg / m2.

[00332] According to one embodiment, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 80 mg / m2.

[00333] In some modalities, ADC can be administered at a dose of approximately 100 mg / m2. Petition 870250084766, dated 09 / 19 / 2025, p. 87 / 149 79 / 115

[00334] In some modalities, ADC can be administered at a dose of approximately 120 mg / m2.

[00335] In some modalities, ADC can be administered at a dose of approximately 150 mg / m2.

[00336] In some modalities, ADC can be administered at a dose of approximately 170 mg / m2.

[00337] According to one embodiment, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 80, 100, 120, 150, or approximately 170 mg / m2 as a loading dose (or first dose).

[00338] According to one embodiment, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 80 mg / m2 as a loading dose.

[00339] According to one embodiment, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 100 mg / m2 as a loading dose.

[00340] According to one embodiment, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 120 mg / m2 as a loading dose.

[00341] According to one embodiment, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 150 mg / m2 as a loading dose.

[00342] According to one embodiment, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 170 mg / m2 as a loading dose. Petition 870250084766, dated 09 / 19 / 2025, p. 88 / 149 80 / 115

[00343] According to one embodiment, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 80, 100, 120, 150 or approximately 170 mg / m2 as a subsequent dose (or second dose).

[00344] According to one embodiment, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 80 mg / m2 as a subsequent dose.

[00345] According to one embodiment, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 100 mg / m2 as a subsequent dose.

[00346] According to one embodiment, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 120 mg / m2 as a subsequent dose.

[00347] According to one embodiment, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 150 mg / m2 as a subsequent dose.

[00348] According to one embodiment, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 170 mg / m2 as a subsequent dose.

[00349] A subsequent dose may be administered on day 1 of cycle(s) following the first cycle (subsequent or additional cycles).

[00350] According to one embodiment, the antibody-drug conjugate comprising the anti-CEACAM5 antibody can be administered at a dose of approximately 80, 100, 120, 150 or approximately 170 Petition 870250084766, dated 09 / 19 / 2025, p. 89 / 149 81 / 115 mg / m2 as a loading dose in a first treatment cycle, for example on day 1, and then a dose of approximately 80, 100, 120, 150 or approximately 170 mg / m2 as a subsequent dose, for example on day 1, in the additional cycle(s).

[00351] In certain embodiments, for individuals 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.

[00352] The antibody-drug conjugate may be tusamitamab ravtansine (huMAb2-3-SPDB-DM4).

[00353] In certain modalities, the ADC is administered approximately once every two weeks. In certain modalities, the ADC is administered approximately once every three weeks. In certain modalities, the ADC is administered approximately once every four weeks. In certain modalities, the ADC is administered approximately once every five weeks. In certain modalities, the ADC is administered approximately once every six weeks. Additional agents

[00354] In certain embodiments, the methods and uses further comprise administering to the individual an effective amount of at least one additional agent effective for treating cancer.

[00355] In certain embodiments, the additional agent is selected from the group consisting of an immune checkpoint inhibitor (ICI), a platinum-based chemotherapy (e.g., cisplatin or carboplatin), pemetrexed, anti-VEGFR2, FOLFOX, FOLFIRI, TAS-102, anti-EGFR, and any combination thereof.

[00356] In certain embodiments, ICI is an anti-PD-1 antibody.

[00357] In certain embodiments, the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, sintilimab, dostarlimab and tislelizumab. Petition 870250084766, dated 09 / 19 / 2025, pp. 90 / 149 82 / 115

[00358] In certain embodiments, the anti-PD-1 antibody is pembrolizumab.

[00359] In certain embodiments, ICI is an anti-PD-L1 antibody.

[00360] In certain embodiments, the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, durvalumab, envafolimab, BMS-936559, CK-301, CS-1001, SHR-1316 (HTI-1088), CBT-502 (TQB-2450) and any combination thereof.

[00361] In certain embodiments, the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab, and durvalumab.

[00362] In certain embodiments, the method comprises administering to the individual an effective amount of tusamitamab ravtansine and pembrolizumab.

[00363] In certain modalities, the method additionally comprises administering to the individual an effective amount of platinum-based chemotherapy.

[00364] In certain modalities, platinum-based chemotherapy is selected from cisplatin and carboplatin.

[00365] In certain embodiments, the method additionally comprises administering an effective amount of pemetrexed to the individual.

[00366] In certain modalities, the method comprises administering to the individual an effective amount of tusamitamab ravtansine, pembrolizumab, and cisplatin.

[00367] In certain embodiments, the method comprises administering to the individual an effective amount of tusamitamab ravtansine, pembrolizumab, cisplatin, and pemetrexed.

[00368] In certain modalities, the method involves administering to the individual an effective quantity of tusamitamabe ra Petition 870250084766, dated 09 / 19 / 2025, pp. 91 / 149 83 / 115 vtansine, pembrolizumab and carboplatin.

[00369] In certain modalities, the method involves administering to the individual an effective amount of tusamitamab ravtansine, pembrolizumab, carboplatin, and pemetrexed.

[00370] In one embodiment, the anti-PD-1 antibody or the anti-PD-L1 antibody, or a fragment thereof, is a monoclonal antibody that has interfering activity with the interaction between PD-1 and PD-L1. In another embodiment, the anti-PD-L1 antibody or the anti-PD-L1 antibody is an IgG.

[00371] Anti-PD-1 and anti-PD-L1 antibodies capable of interfering with the 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 capable and ready to fight cancer. When PD-1 and PD-L1 interact, they form a biochemical shield that protects tumor cells from being destroyed by the immune system. Thus, blocking PD-1 or PD-L1, leading to the blocking of the interaction between PD-1 and PD-L1, prevents or unmasks the biochemical shield that protects tumor cells from being destroyed by the immune system.

[00372] Some 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.

[00373] Similarly, some anti-PD-L1 antibodies have been approved for clinical use in the treatment of cancer. These include atezolizumab (TECENTRIQ®), avelumab (BAVENCIO®) and durvalumab (IMFINZI®).

[00374] In one embodiment, the anti-PD-1 antibody is pembrolized Petition 870250084766, dated 09 / 19 / 2025, pp. 92 / 149 84 / 115 mabe or sintilimabe.

[00375] In one embodiment, the anti-PD-1 antibody is pembrolizumab. It is a fully human monoclonal IgG1 antibody against human PD-1.

[00376] In one embodiment, the anti-PD-1 antibody, or a fragment thereof, comprises the light chain and heavy chain CDRs of pembrolizumab.

[00377] In one embodiment, the anti-PD-1 antibody or a fragment thereof comprises the variable heavy chain (VH) domain and the variable light chain (VL) domain of pembrolizumab.

[00378] 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 heavy chain (VH) domain and the variable light chain (VL) domain of sintilimab.

[00379] 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) comprising 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 a light chain that may or may not contain a CH1 domain.

[00380] 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.

[00381] The anti-PD-1 antibody or the anti-PD-L1 antibody may also be an antibody fragment, in particular, a fragment of Petition 870250084766, dated 09 / 19 / 2025, pp. 93 / 149 85 / 115 humanized antibody selected from the group consisting of Fv, Fab, F(ab')2, Fab', dsFv, (dsFv)2, scFv, sc(Fv)2 and diacorpos.

[00382] The antibody may also be a bispecific or multispecific antibody formed from antibody fragments, wherein at least one antibody fragment is an antibody fragment according to the invention. The anti-PD-1 antibody or the anti-PD-L1 antibody and fragments thereof may be produced by any technique well known in the art. In particular, said antibodies are produced by techniques as already described.

[00383] The anti-PD-1 antibody or the anti-PD-L1 antibody and fragments thereof may be used in an isolate (e.g., purified) from or contained in a vector, such as a membrane or lipid vesicle (e.g., a liposome).

[00384] Anti-PD-1 antibody or 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 alone or in combination.

[00385] 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.

[00386] In one embodiment, the anti-VEGFR-2 antibody is a monoclonal antibody or a fragment thereof that has antagonistic activity toward VEGFR-2. In another embodiment, the anti-VEGFR-2 antibody is an IgG.

[00387] The anti-VEGFR-2 antibody is preferably adapted to the patient. For example, an anti-murine VEGFR-2 antibody, such as DC-101, is preferably used in mice, and an anti-human VEGFR-2 antibody in humans.

[00388] In one embodiment, the anti-VEGFR-2 antibody is ramucirumab (CAS number 947687-13-0). The same is an antibody of Petition 870250084766, dated 09 / 19 / 2025, pp. 94 / 149 86 / 115 Fully human monoclonal IgG1 antibody against human VEGFR-2.

[00389] In one embodiment, the anti-VEGFR-2 antibody or fragment thereof comprises the light chain and heavy chain CDRs of ramucirumab.

[00390] In one embodiment, the anti-VEGFR-2 antibody or fragment thereof comprises the variable heavy chain (VH) domain and the variable light chain (VL) domain of ramucirumab.

[00391] The anti-VEGFR-2 antibody or 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) comprising 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 a light chain that may or may not contain a CH1 domain.

[00392] 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.

[00393] The anti-VEGFR-2 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 diacorpos.

[00394] The antibody may also be a bispecific or multispecific antibody formed from antibody fragments, wherein at least one antibody fragment is an antibody fragment according to the invention. The anti-VEGFR-2 antibody and fragments thereof may be produced by any technique well known in the art. In particular, said antibodies are produced by techniques as already described. Petition 870250084766, dated 09 / 19 / 2025, pp. 95 / 149 87 / 115

[00395] The anti-VEGFR-2 antibody and fragments thereof can be used in an isolate (e.g., purified) from or contained in a vector, such as a membrane or lipid vesicle (e.g., a liposome).

[00396] Anti-VEGFR-2 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 alone or in combination.

[00397] The immunoconjugate comprising an anti-CEACAM5 antibody should be used in combination with cetuximab for the treatment of cancer.

[00398] Cetuximab (CAS number 205923-56-4) is a chimeric monoclonal IgG1 antibody against the epidermal growth factor receptor (EGFR). Cetuximab itself has been used for the treatment of metastatic colorectal cancer, metastatic non-small lung cancer, and head and neck cancer.

[00399] The immunoconjugate comprising an anti-CEACAM5 antibody should be used in combination with TAS-102 for the treatment of cancer.

[00400] TAS-102 is a known chemotherapy regimen approved for human use that comprises the combined administration of trifluoridine and tipiracil and is typically administered in 4-week cycles. TAS-102 combines trifluoride and tipiracil and has been used in the treatment of colorectal cancer.

[00401] As a modified deoxyuridine, trifluoridine (CAS registry number 70-00-8) is a nucleoside analog that is incorporated into DNA. The modified DNA binds to thymidylate synthase, inhibiting the enzyme's activity. Tipiracil (CAS registry number 183204-742) is a thymine analog that prevents the degradation of trifluoridine by thymidine phosphorylase. Petition 870250084766, dated 09 / 19 / 2025, pp. 96 / 149 88 / 115

[00402] The immunoconjugate comprising an anti-CEACAM5 antibody should be used in combination with FOLFIRI for the treatment of cancer.

[00403] FOLFIRI itself is a known chemotherapy regimen approved for human use that comprises the combined administration of folinic acid, 5-fluorouracil, and irinotecan, and is typically administered in up to 12 two-week cycles. FOLFIRI combines drugs, each with a different mechanism of action and favorably synergistic effects, causing the death of cancer cells.

[00404] 5-Fluoro-uracil (CAS registry number 51-21-8) is an antimetabolite that primarily inhibits thymidylate synthase and thus blocks thymidine synthesis. 5-Fluoro-uracil has been used in the treatment of colon cancer, esophageal cancer, stomach cancer, pancreatic cancer, breast cancer, and cervical cancer.

[00405] Folinic acid, also known as leucovorin (CAS registry number 58-05-9), stabilizes the complex between 5-fluorouracil and thymidylate synthase, increasing the cytotoxicity of 5-fluorouracil. In one embodiment, folinic acid is L-folinic acid (N-[4-[[[(6S)-2-amino-5-formyl-3,4,5,6,7,8-hexahydro-4-oxo-6-pteridinyl]methyl]amino]benzoyl]-L-glutamic acid). In another embodiment, folinic acid is the calcium salt of L-folinic acid. Folinic acid may also comprise a mixture of two or more stereoisomers.

[00406] Irinotecan (CAS number 97682-44-5) is a cytotoxic agent that is a semi-synthetic derivative of the alkaloid camptothecin and inhibits topoisomerase I, resulting in the inhibition of DNA replication and transcription, and has been used in the treatment of colon cancer and small cell lung cancer.

[00407] The immunoconjugate comprising an anti-CE antibody Petition 870250084766, dated 09 / 19 / 2025, pp. 97 / 149 89 / 115 ACAM5 should be used in combination with FOLFOX for the treatment of cancer.

[00408] FOLFOX itself is a known chemotherapy regimen approved for human use that comprises the combined administration of folinic acid, 5-fluorouracil, and oxaliplatin and is typically administered in up to 12 two-week cycles. FOLFOX combines drugs, each with a different mechanism of action and favorably synergistic effects, causing the death of cancer cells.

[00409] 5-Fluoro-uracil (CAS registry number 51-21-8) is an antimetabolite that primarily inhibits thymidylate synthase and thus blocks thymidine synthesis. 5-Fluoro-uracil has been used in the treatment of colon cancer, esophageal cancer, stomach cancer, pancreatic cancer, breast cancer, and cervical cancer.

[00410] Folinic acid, also known as leucovorin (CAS registry number 58-05-9), stabilizes the complex between 5-fluorouracil and thymidylate synthase, increasing the cytotoxicity of 5-fluorouracil. In one embodiment, folinic acid is L-folinic acid (N-[4-[[[(6S)-2-amino-5-formyl-3,4,5,6,7,8-hexahydro-4-oxo-6-pteridinyl]methyl]amino]benzoyl]-L-glutamic acid). In another embodiment, folinic acid is the calcium salt of L-folinic acid. Folinic acid may also comprise a mixture of two or more stereoisomers.

[00411] Oxaliplatin (CAS number 61825-94-3) is known to cross-link DNA strands, preventing DNA replication and transcription, and has been used in the treatment of colorectal cancer. Cancer

[00412] In one embodiment, the cancer is a carcinoma, a sarcoma, or a blastoma. In an additional embodiment, the cancer is a carcinoma. Petition 870250084766, dated 09 / 19 / 2025, pp. 98 / 149 90 / 115

[00413] According to one classification, the cancer is a cancer that expresses CEACAM5. A cancer that expresses CEACAM5 may also be named CEACAM5-positive cancer.

[00414] In some modalities, the cancer is a CEACAM5 positive cancer.

[00415] A CEACAM5-positive cancer is defined as a cancer for which an immunohistochemical [IHC] intensity of CEACAM5 is > 2+ in > 50% of cancer cells or intensity of > 2+ in > 1% and < 50% of tumor cells (or cancer cells).

[00416] In certain modalities, cancer has negative or low expression of CEACAM5 in tumor cells. Negative or low expression of CEACAM5 in tumor cells is defined as an immunohistochemical [IHC] intensity of CEACAM5 of > 2+ in < 1% of cells, as measured by immunohistochemistry (IHC).

[00417] In certain modalities, cancer that has moderate expression for CEACAM5 in tumor cells. Moderate expression of CEACAM5 in tumor cells can be defined as having an immunohistochemical intensity [IHC] of CEACAM5 of > 2+ in > 1% and in < 50% of cancer cells as measured by immunohistochemistry.

[00418] In certain modalities, cancer has a high expression of CEACAM5 in tumor cells. A high expression of CEACAM5 in tumor cells can be defined as having an immunohistochemical intensity [IHC] of CEACAM5 of >2+ in > 50% of cancer cells, as measured by immunohistochemistry.

[00419] Immunohistochemical techniques for detecting antigens in cells or tissue sections through immunological and chemical reactions are well known in the field. These techniques are highly sensitive and specific and can detect a wide variety of antigens. Petition 870250084766, dated 09 / 19 / 2025, pp. 99 / 149 91 / 115 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 an enzyme-conjugated secondary antibody and generation of colored deposits at the antibody-antigen binding sites in the presence of substrate and chromogenic enzyme catalyzed by the enzyme.

[00420] Tumor expression of CEACAM5 can be determined using immunohistochemistry (IHC) assay. An assay can be performed using anti-CEACAM5 antibody, such as SANOFI clone 769. Anti-CEACAM5 clone 769 is a murine monoclonal antibody with the same specificity as tusamitamab ravtansine for the CEACAM5 target. The assay can be performed on a Techmate platform or on a Dako / Agilent Autostainer Link 48 IHC or any other immunohistochemistry platforms. Interpretation of CEACAM5 reactivity is performed using semi-quantitative Percent Scores (calculated by summing the percentages of intensities >2+) or H-score for plasma membrane staining of CEACAM5 (whole or polarized) in tumor cells.

[00421] Depending on the modality, the cancer is selected from among hepatocellular carcinoma, colorectal cancer, gastric cancer, adenocarcinoma of the gastroesophageal junction (GEJ), esophageal cancer, lung cancer (e.g., non-small cell non-squamous lung cancer), cervical cancer, pancreatic cancer, ovarian cancer, thyroid cancer, bladder cancer, endometrial cancer, breast cancer, liver cancer, biliary tract cancer (e.g., cholangiocarcinoma), prostate cancer, or skin cancer.

[00422] The cancer may be selected from among colorectal cancer, gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, esophageal cancer, pancreatic cancer, and lung cancer. Petition 870250084766, dated 09 / 19 / 2025, pp. 100 / 149 92 / 115

[00423] In some forms, the cancer may be colorectal cancer.

[00424] In some forms, the cancer may be pancreatic cancer.

[00425] In some modalities, the cancer can be selected from among gastric cancer, adenocarcinoma of the gastroesophageal junction (GEJ), esophageal cancer, and lung cancer.

[00426] In some forms, the cancer may be gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, or esophageal cancer.

[00427] According to one classification, the cancer is gastric cancer or adenocarcinoma of the gastroesophageal junction (GEJ).

[00428] According to one theory, the cancer is gastric cancer.

[00429] According to one theory, the cancer is lung cancer.

[00430] Lung cancer can be non-small cell non-squamous lung cancer (NSQNSCLC).

[00431] Non-small cell lung cancer is a disease in which malignant (cancerous) cells form in the tissues of the lung. Smoking is the main cause of the disease. This is a type of epithelial lung cancer different from small cell lung carcinoma. There are several types of non-small cell lung cancer. Each type of non-small cell lung cancer has different types 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 according to the types of cells present in the cancer and how the cells appear under a microscope: (1) squamous cell carcinoma: Cancer that begins in squamous cells, which are thin, flat cells resembling... Petition 870250084766, dated 09 / 19 / 2025, pp. 101 / 149 93 / 115 tes a fish scales. Also called squamous cell carcinoma. (2) large cell carcinoma: Cancer that can start in several types of large cells. (3) adenocarcinoma: Cancer that starts in the cells that line the alveoli and make up substances like mucus.

[00432] In some forms, non-small cell non-squamous lung cancer may be an advanced or metastatic NSCLC NSQ.

[00433] According to one modality, the individual is a patient with a malignant tumor, in particular, with a solid malignant tumor, and more specifically, with a locally advanced or metastatic solid malignant tumor. A metastatic solid malignant tumor can be a metastatic cancer, for example, a metastatic carcinoma. A cancer or a carcinoma can be as indicated above.

[00434] In some forms, non-small cell non-squamous lung cancer does not have a sensitizing mutation of the epidermal growth factor receptor (EGFR) or a mutation of the B1 homolog of the murine sarcoma virus oncogene v-raf (BRAF) or alterations in anaplastic lymphoma kinase / c-ros 1 oncogene (ALK / ROS). Pharmaceutical compositions or combinations

[00435] In some embodiments, in the uses and methods as disclosed in this document, the administration of ADC may be performed via the parenteral route. A suitable parenteral route may be intravenous infusion.

[00436] The present invention also relates to an ADC for the manufacture of a cancer treatment drug that the combination comprises.

[00437] In some embodiments, the present invention relates to a pharmaceutical composition comprising (i) an ADC as disclosed herein, and a pharmaceutically available excipient. Petition 870250084766, dated 09 / 19 / 2025, pp. 102 / 149 94 / 115 acceptable.

[00438] The ADC of the invention can be combined with pharmaceutically acceptable excipients, and optionally sustained-release matrices, such as biodegradable polymers, to form therapeutic compositions.

[00439] Thus, another object of the invention relates to a pharmaceutical composition comprising an ADC of the invention and a pharmaceutically acceptable carrier or excipient. ADC or an immunoconjugate according to the invention, for use as a medicament.

[00440] The invention also relates to an ADC according to the invention, for use in the treatment of cancer.

[00441] Pharmaceutical excipient or pharmaceutically acceptable excipient refers to molecular entities and compositions that do not produce an adverse, allergic or other undesirable reaction when administered to a mammal, especially a human being, as appropriate. A carrier or pharmaceutically acceptable excipient refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation aid of any kind.

[00442] As used in this document, pharmaceutically acceptable carriers or excipients include any and all solvents, dispersing 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 the following: water, amino acids, saline solution, phosphate-buffered saline solution, phosphate buffer, acetate, citrate, succinate; amino acids and derivatives, such as histidine, arginine, glycine, proline, glycylglycine; inorganic salts such as NaCl, calcium chloride; sugars or polyalcohols such as dextrose, glycerol, ethanol, sucrose, trehalose, mannitol; surfactants such as Polysorbate 80, Polysorbate 20, Poloxamer 188 and Petition 870250084766, dated 09 / 19 / 2025, pp. 103 / 149 95 / 115 similar as well as a combination thereof. In many cases, it will be preferable to include isotonic agents, such as sugars, polyalcohols or sodium chloride in the composition, and the formulation may also contain an antioxidant, such as tryptamine, and a stabilizing agent, such as Tween 20.

[00443] The form of pharmaceutical compositions, the route of administration, the dosage and the regimen naturally depend on the condition being treated, the severity of the disease, the age, weight and sex of the individual, etc.

[00444] The pharmaceutical compositions of the invention can be formulated for topical, oral, parenteral, intranasal, intravenous, intramuscular, subcutaneous or intraocular administration and the like. In one embodiment, the pharmaceutical compositions and combinations of the invention are formulated for intravenous administration.

[00445] In particular, pharmaceutical compositions contain vehicles or excipients that are pharmaceutically acceptable for a formulation capable of being injected. These may be, in particular, sterile isotonic saline solutions (monosodium or disodium phosphate, sodium, potassium, calcium or magnesium chloride and similar substances or mixtures of such salts), or dry compositions, especially frozen ones, which, after the addition, depending on the case, of sterile water or physiological saline, allow the formation of injectable solutions.

[00446] The pharmaceutical composition can be administered via drug combination devices.

[00447] The doses used for administration can be adapted as a function of several parameters, and in particular as a function of the method of use, the relevant pathology or, alternatively, the desired duration of treatment.

[00448] To prepare pharmaceutical compositions, a quantity Petition 870250084766, dated 09 / 19 / 2025, pp. 104 / 149 96 / 115 effective antibody-drug conjugate comprising an anti-CEACAM5 antibody and / or an anti-CTLA4 antibody and / or an anti-PD-1 or anti-PD-L1 antibody may be dissolved or dispersed in a pharmaceutically acceptable carrier or aqueous medium.

[00449] Suitable pharmaceutical forms for injection include sterile aqueous solutions or dispersions; formulations including sesame oil, peanut oil, or aqueous propylene glycol; and sterile powders for 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 administration without degradation. It must be stable under manufacturing and storage conditions and must be preserved against the contaminating action of microorganisms such as bacteria and fungi.

[00450] 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 common storage and use conditions, these preparations contain a preservative to prevent the growth of microorganisms.

[00451] The antibody-drug conjugate comprising an anti-CEACAM5 antibody may be formulated in a neutral or salt form. Pharmaceutically acceptable salts include acid addition salts (formed with the free amino groups of the protein) and those formed with inorganic acids, such as hydrochloric or phosphoric acids, or organic acids, such as acetic, oxalic, tartaric, mandelic and the like. Salts formed with free carboxyl groups may also be derived from inorganic bases, such as sodium, potassium, ammonium, calcium or ferric hydroxides, and organic bases such as isopropylamine, trimethylammonium chloride, and methylammonium chloride. Petition 870250084766, dated 09 / 19 / 2025, pp. 105 / 149 97 / 115 lamin, glycine, histidine, procaine and the like.

[00452] The carrier may also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol and the like), suitable mixtures thereof, and vegetable oils. Appropriate fluidity may be maintained, for example, by the use of a coating such as lecithin, by the subsequent particle size required in the case of dispersion, and by the use of surfactants. Prevention of microbial action may be achieved 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 injectable compositions may be achieved by the use in the compositions of absorption-retardant agents, for example, aluminum monostearate and gelatin.

[00453] Sterile injectable solutions are prepared by incorporating the active compounds in the required quantity into the appropriate solvent with several of the other ingredients listed above as needed, followed by sterilized filtration. In general, dispersions are prepared by incorporating the various sterilized active ingredients into a sterilized vehicle containing the basic dispersion medium and the other necessary ingredients from those listed above. In the case of sterilized powders for the preparation of sterile injectable solutions, the preferred preparation methods are vacuum drying and lyophilization techniques that produce a powder of the active ingredient plus any desired additional ingredient from a previously sterilely filtered solution.

[00454] The preparation of more concentrated or highly concentrated solutions for direct injection is also contemplated, in which the Petition 870250084766, dated 09 / 19 / 2025, pp. 106 / 149 98 / 115 The use of DMSO as a solvent is predicted to result in extremely rapid penetration, delivering high concentrations of the active agents to a small tumor area.

[00455] After formulation, the solutions will be administered in a manner consistent with the dosage formulation and in such a quantity as to be therapeutically effective. The formulations are readily administered in a variety of dosage forms, with the injectable solutions described above, but drug-release capsules and similar forms may also be used.

[00456] For parenteral administration in an aqueous solution, for example, the solution should be adequately buffered, if necessary, and the liquid diluent should first be made isotonic with sufficient saline or glucose solution. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous, and intraperitoneal administration. In this connection, sterile aqueous media that can be used will be known to those skilled in the art in light of the present invention. For example, a dosage may be dissolved in 1 ml of isotonic NaCl solution and added to 1000 ml of hypodermoclysis fluid or injected into the proposed infusion site (see, for example, Remington's Pharmaceutical Sciences 15th Edition, pages 1035-1038 and 1570-1580). Some variation in dosage will necessarily occur depending on the condition of the individual to be treated.The person responsible for administration will determine, in any case, the appropriate dose for the individual.

[00457] The antibody-drug conjugate comprising an anti-CEACAM5 antibody formulated for parenteral administration, such as intravenous or intramuscular injection; other pharmaceutically acceptable forms include, for example, tablets or other solids for oral administration; prolonged-release capsules; and Petition 870250084766, dated 09 / 19 / 2025, pp. 107 / 149 99 / 115 or any other form currently in use.

[00458] 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 skilled in the art.

[00459] Nanocapsules can generally retain compounds in a stable and reproducible manner. To avoid side effects due to intracellular polymer overload, such ultrafine particles (sized around 0.1 μm) are generally designed using polymers that have the ability to be degraded in vivo. Biodegradable polyalkyl cyanoacrylate nanoparticles or biodegradable polylactide or polylactide coglycolide nanoparticles that meet these requirements are contemplated for use in the present invention, and such particles can be easily produced.

[00460] Liposomes are formed from phospholipids that are dispersed in an aqueous medium and spontaneously form concentric multilamellar double-layer vesicles (also called multilamellar vesicles (MLVs)). MLVs generally have diameters of 25 nm to 4 μm. Sonication of MLVs results in the formation of small unilamellar vesicles (SUVs) with diameters in the range of 200 to 500 Å, containing an aqueous solution in the core. The physical characteristics of liposomes depend on pH, ionic strength, and the presence of divalent cations. Administration Methods and Formulations

[00461] The methods described herein comprise the administration of a therapeutically effective amount of an antiCEACAM5 ADC to an individual. As used herein, an effective amount or therapeutically effective quantity is a dose of the therapeutic agent that results in treatment of a cancer expressing CEACAM5 (e.g., lung cancer, gastric cancer, junctional cancer). Petition 870250084766, dated 09 / 19 / 2025, pp. 108 / 149 100 / 115 gastroesophageal, or esophageal cancer). As used herein, treat refers to causing a detectable improvement in one or more symptoms associated with cancer expressing CEACAM5 (e.g., lung cancer) or causing a biological effect (e.g., a decrease in the level of a particular biomarker) that is correlated with the underlying pathological mechanism(s) that give rise to the condition or symptom(s). For example, a dose of antiCEACAM5 ADC that causes an improvement in any of the following symptoms or conditions associated with cancer expressing CEACAM5 is considered a therapeutically effective amount.

[00462] In another example, a treatment was not effective when a dose of anti-CEACAM5 ADC does not produce a detectable improvement in one or more parameters or symptoms associated with a cancer expressing CEACAM5 (e.g., lung cancer, gastric cancer, gastroesophageal junction cancer, or esophageal cancer) or does not produce a biological effect that is correlated with the underlying pathological mechanism(s) that gives rise to the cancer condition or symptom(s).

[00463] According to some of these modalities, the ADC antiCEACAM5 is administered intravenously.

[00464] According to the methods of the present invention, a therapeutically effective amount of anti-CEACAM5 ADC that is administered to the individual will vary depending on the age and stature (e.g., body weight or body surface area) of the individual as well as the route of administration and other factors well known to those of common skill in the art.

[00465] In certain modalities, the dose of ADC varies depending on the individual's body surface area. In certain modalities, the dose of anti-CEACAM5 ADC administered to the individual ranges from approximately 1 mg / m2 to approximately 500 mg / m2. In some modalities, the Petition 870250084766, dated 09 / 19 / 2025, pp. 109 / 149 The dose of ADC administered to an individual ranges from approximately 5 mg / m2 to approximately 300 mg / m2. In various embodiments, the dose of ADC administered to an individual ranges from approximately 5 mg / m2 to approximately 250 mg / m2. In various embodiments, the dose of ADC administered to an individual ranges from approximately 60 mg / m2 to approximately 190 mg / m2. In various embodiments, the dose is approximately 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, or 210 mg / m2 based on the individual's body surface area. In certain modalities, the ADC dose is approximately 100 mg / m2. In certain modalities, the ADC dose is approximately 150 mg / m2. In certain modalities, the ADC dose is approximately 170 mg / m2. In certain modalities, the ADC dose is approximately 190 mg / m2.

[00466] In several modalities, the ADC dose is 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, or 210 mg / m2 based on the individual's body surface area. In certain modalities, the ADC dose is 100 mg / m2. In certain modalities, the ADC dose is 150 mg / m2. In certain modalities, the ADC dose is 170 mg / m2. In certain modalities, the ADC dose is 190 mg / m2. [EXAMPLES]

[00467] The following examples illustrate embodiments of the invention that are currently best known. However, it should be understood that the following content serves only to exemplify or illustrate the application of the principles of the present invention. Numerous modifications and compositions, methods and alternative systems may be devised by persons skilled in the art without departing from the spirit and scope of the present invention. Thus, although the present invention has been described above in particular, the following examples provide further details in connection with what are presently considered to be the most practical embodiments and Petition 870250084766, dated 09 / 19 / 2025, pp. 110 / 149 102 / 115 preferred aspects of the invention. EXAMPLE 1: Materials & Methods Study Design and Patients

[00468] The study design details for this first-ever open-label, phase 1 / 2 human dose escalation and expansion trial have been published elsewhere.

[15] Ethics statement

[00469] In summary, patients eligible for the dose escalation and expansion phases were >18 years of age with locally advanced or metastatic solid tumors for which no standard alternative therapy was available, and an Eastern Cooperative Oncology Group (ECOG PS) performance status of 0 or 1.

[00470] The dose-escalation phase population was enriched, but not restricted, to patients with tumor types that express CEACAM5, and expression was retrospectively confirmed using IHC on the most recent archival tissue samples.

[00471] The expanding population was restricted to separate cohorts of patients with advanced colorectal cancer, NSQ-NSCLC, small cell lung cancer, and gastric adenocarcinoma.

[00472] In this report, we present results only for patients with NSQ-NSCLC. There were 2 expansion phase populations of NSQ-NSCLC, based on IHC analysis of tumor tissue:

[00473] High CEACAM5 expressers defined as patients with CEACAM5 expression at an intensity > 2+ in > 50% of the tumor cell population and

[00474] Moderate CEACAM5 expressers defined as patients with CEACAM5 expression at an intensity > 2+ in > 1% to < 50% of the tumor cell population.

[00475] It was necessary for all participants to have at least one measurable injury according to Response Assessment Criteria. Petition 870250084766, dated 09 / 19 / 2025, pp. 111 / 149 103 / 115 in Solid Tumors (RECIST) v1.1.

[00476] The main exclusion criteria included: life expectancy < 12 weeks; known or symptomatic brain metastases; receiving other cancer treatment; having received prior therapy targeting CEACAM5; having received prior treatments with maitansanoid; and poor bone marrow reserve or organ dysfunction. Treatments

[00477] During the expansion phase, all patients received intravenous (IV) tusamitamab ravtansine 100 mg / m2 Q2W, as determined during the dose escalation phase of this study

[15] . Tusamitamab ravtansine was infused at 2.5 mg / min for 30 minutes, then 5 mg / min, provided there were no signs or symptoms of a hypersensitivity reaction.

[00478] To prevent hypersensitivity reactions, patients were premedicated with an oral antihistamine 1 hour before receiving tusamitamab ravtansine.

[00479] Treatment was continued until disease progression, unacceptable toxicity, or the patient's willingness to stop. Results

[00480] The primary outcome during the expansion phase assessed the objective response rate according to the RECIST v1.1 criteria.

[00481] The main secondary results assessed safety, which will be reported in a separate publication.

[00482] The exploratory biomarker objectives (reported here) included exploring the potential link between CEACAM5 expression characteristics and response (expansion phase), investigating potential biomarkers (other than CEACAM5) that may predict tusamitamab ravtansine activity (expansion phase), and assessing the potential of circulating CEA levels to be a diagnostic tool. Petition 870250084766, dated 09 / 19 / 2025, pp. 112 / 149 104 / 115 co-adjuvant suitable for treatment with tusamitamitamab ravtansine (evaluating the correlation between circulating CEA levels and tumor CEACAM5 expression and the correlation between this biomarker and treatment response). Tumor samples

[00483] Tumor samples were obtained using the most recent archived tumor samples (i.e., tumor tissue archived at diagnosis, surgery, or collected before the patient's inclusion in the study and not undergoing cancer treatment); fresh baseline biopsy was optional in patients who had a biopsy-appropriate lesion. CEACAM5 expression by immunohistochemistry

[00484] The level and pattern of CEACAM5 expression in archival tumor samples were analyzed by IHC locally at clinical sites and / or centrally in a laboratory using mouse anti-CEACAM5 clone 769, which has the same specificity as tusamitamab ravtansine for the CEACAM5 target.

[00485] At least six 5 μm slides plus three additional 10 μm slides (or six 5 μm slides) of formalin-fixed paraffin-embedded tissue (FFPE) should be provided for each patient. CEACAM5 expression was centrally determined using a validated IHC assay with the 769 clone anti-CEACAM5 antibody at 0.5 μg / ml.

[00486] The reactivity of CEACAM5 was evaluated in tumor cells using semi-quantitative Percent Scores (calculated by summing the percentages of intensities >2+) or H-score for CEACAM5 plasma membrane staining (whole or polarized); CEACAM5 cytoplasmic staining was also evaluated (centrally only). RNA / DNA residual tissue extraction Petition 870250084766, dated 09 / 19 / 2025, pp. 113 / 149 105 / 115

[00487] If blocks were provided, FFPE tumor tissues were sectioned to 10 μm thickness using a microtome, and 3 sections were mounted on adhesive microscope slides. After trimming excess paraffin from the sample slide using a sterile scalpel, the tumor tissue was macrodissected and collected in individual Eppendorf DNA LoBind Tubes. Genomic DNA and total RNA from each human lung cancer FFPE tissue sample were co-extracted using the ALLPREP® DNA / RNA FFPE kit (ref. 80234, QIAGEN), according to the manufacturer's instructions, with initial manual processing steps followed by final automated extraction steps using the QIACUBE automated nucleic acid purification instrument. Genomic DNA was eluted in ATE buffer at 30 μl and total RNA eluted in RNase-free water at 20 μl.

[00488] A total of 71 RNA samples were generated for RNA sequencing analysis. Gene expression (RNA-seq)

[00489] Tissue samples with known CEACAM5 expression levels by IHC were analyzed using RNA-seq. RNA samples from 57 biopsies of FFPE patients were sequenced using the KAPA mRNA Platform HYPERPREPKITILLUMINA®. RNA-seq data were processed as follows: sequencing reads were mapped to the GRCh 38 reference genome using Reference Alignment of Spliced ​​Transcripts (STAR)

[25] . Gene expression was initially measured in FPKM (Fragments Per Million Kilobases) by CUFFLINK

[26] and gene-level FFKM was converted to TPM (Transcripts Per Million Kilobases)

[27] . TPM values ​​were log2 transformed and quantile normalized for downstream analysis, including differential gene expression (DGE) analysis. Samples with fewer than 10,000 detected genes were excluded from the analysis. Petition 870250084766, dated 09 / 19 / 2025, pp. 114 / 149 106 / 115 downstream. RNA-seq included counter-analysis of microenvironment cell populations [MCP], according to published methods [28, 29]. Statistical Analysis

[00490] All analyses were performed on the biomarker population, defined as patients who received treatment and who had at least one evaluable CEACAM5 expression measurement or at least one valid RNA assessment. Expression of CEACAM5 in pre-treatment tumor samples

[00491] The percentage of tumor cells positive in intensity >2+ on the overall membrane defined the expression of CEACAM5 in the tumor tissue, which was analyzed using descriptive statistics.

[00492] Associations between CEACAM5 expression characteristics were assessed by calculating P values ​​from Mann-Whitney U tests (when only 2 groups were present) or Kendall tau tests.

[00493] The adjusted P-values ​​were also calculated using the Benjamini-Hochberg (BH) multiple correction procedure to control for the false discovery rate. Association between baseline biomarkers and tumor response

[00494] The statistical significance of the association between CEACAM5 expression (moderate versus high expression) and overall response rate (ORR) was assessed using a two-tailed Fisher's exact test. EXAMPLE 2: Results Biomarker-evaluable patients

[00495] In this exploratory biomarker analysis of the dose-expansion phase cohort of patients with advanced non-squamous NSCLC, the first patient was enrolled on January 2, 2017, and the data cutoff for these analyses was December 2020.

[00496] Of the 888 patients pre-screened with NSQ-NSCLC, 172 Petition 870250084766, dated 09 / 19 / 2025, pp. 115 / 149 107 / 115 (19%) showed high expression of CEACAM5 and 210 (24%) had moderate expression of CEACAM5.

[00497] 92 patients were treated: 64 with high expression of CEACAM5 (high expressers) and 28 with moderate expression of CEACAM5 (moderate expressers)

[00498] In summary, the median age was 62.5 years (range, 31-91 years; 42% were >65 years of age); 51% were male; 72% had ECOG PS >1; patients had received a median of 3 prior lines of therapy (range, 1-10 lines) for advanced disease, including anti-tubalin agents (61%) and anti-PD1 / PD-L1 agents (75%).

[00499] Clinical findings showed that there was an enrichment of clinical responses in patients (responders) with high levels of CEACAM5 protein expression measured by immunohistochemistry (IHC), i.e., at an intensity > 2+ in > 50% of tumor cells measured by IHC for CEACAM5. Non-responders were moderate expressers of CEACAM5 protein, defined as CEACAM5 protein expression measured by IHC at an intensity of > 2+ in > 1% to less than 50% of the tumor cell population. CEACAM5 expression: staining pattern and distribution in tumors

[00500] High CEACAM5 expressers showed a predominance of whole-membrane expression over polarized expression. In contrast, whole-membrane and polarized expression was similar among moderate CEACAM5 expressers. (Table 1).

[00501] At initial diagnosis, the predominant histological type was adenocarcinoma and most (91.3%) of tumors expressing CEACAM5 were stage III or higher, regardless of the CEACAM5 expression level (Table 1). Petition 870250084766, dated 09 / 19 / 2025, pp. 116 / 149 108 / 115 Table 1. CEACAM5 expression: staining pattern and distribution in tumors. Percentage of CEACAM5 positive cells: high and moderate intensity (N = 64) (N = 28) Color pattern, mean ± SD Whole Polarized Membrane 51.95 ±37.43 25.08 ±31.69 6.96 ± 9.02 6.89 ± 12.04 Histology Type, n (%) Adenocarcinoma 91 (98.9) 63 (98.4) 28 (100) Other 1 (1.1) 1 (1.6) 0 Staging, n (%) Stage I 6 (6.5) 5 (7.8) 1 (3.6) Stage II 2 (2.2) 2 (3.1) 0 Stage III 20 (21.7) 14 (21.9) 6 (21.4) Stage IV 64 (69.6) 43 (67.2) 21 (75.0) aN = 63 for polarized membrane.

[00502] CEACAM5, carcinoembryonic antigen-related cell adhesion molecule 5; NSCLC, non-small cell lung cancer; SD, standard deviation. Associations between CEACAM5 expression by IHC and gene expression (RNA-seq)

[00503] Differential gene expression analysis identified CEACAM5 mRNA as the gene most strongly associated (and the only significant adjusted P = 0.00265) with high vs. moderate CEACAM5 expression by IHC. Expression of another member of the CEACAM family or other gene expression is not significantly associated with CEACAM5 IHC after correction for multiple tests (Table 2). Petition 870250084766, dated 09 / 19 / 2025, pp. 117 / 149 109 / 115 Table 2. Fold changes in gene expression by RNA-seq in high versus moderate expression of CEACAM5 by IHC: The top 10 ranking genes, including the closest family member of CEACAM. Classification Gene logFC Lower CL Upper CL AveExpr P-value Adjusted P-value 1 CEACAM5 0.46 0.28 0.63 7.1 3.2e-06 0.05 2 TACR1 -0.96 -1.4 -0.51 4.2 6.8e-05 0.38 3 HOXB5 1 0.54 1.5 5.3 7.3e-05 0.38 4 RIOX2 -0.15 -0.22 -0.071 6.5 0.00024 0.92 5 SGCE -0.22 -0.33 -0.1 6.4 0.00034 1 6 HOXB6 0.82 0.38 1.3 5.2 0.00045 1 7 ENTPD8 1.2 0.53 1.9 4.8 0.00069 1 8 MNX1 0.99 0.44 1.5 4.6 0.00072 1 9 CYP4Z1 -0.93 -1.4 -0.41 4.2 0.00076 1 10 RASL11A -0.55 -0.86 -0.24 5.5 0.00077 1 CEACAM3 0.65 0.13 1.2 6.3 0.016 1 CEACAM, carcinogen-related cell adhesion molecule; CL, confidence limit; logFC, log fold change. Associations between CEACAM5 expression, objective response rate, and CEACAM5 mRNA level.

[00504] A correlation was observed between CEACAM5 mRNA level and CEACAM5 expression measured by immunohistochemistry recorded as the sum of the percentage of tumor cells expressing target intensity at least 2+ in patients who responded to treatment with tusamitamab ravtansine. EXAMPLE 3: Discussion

[00505] Nearly 20% of pre-screened NSCLC patients showed high CEACAM5 expression, as measured by IHC, i.e., staining intensity > 2+ in > 50% of tumor cells. An enrichment of clinical responses was observed in patients with Petition 870250084766, dated 09 / 19 / 2025, pp. 118 / 149 110 / 115 a high level of expression of the CEACAM5 protein and who are being treated with tusamitamab ravtansine.

[00506] An association was found between the expression of CEACAM5, cCEA, cCEACAM5 and tumor mRNA levels of CEACAM5.

[00507] Furthermore, CEACAM5 mRNA expression was significantly upregulated in patients with high vs. moderate CEACAM5 protein expression, but other genes, including other genes in the CEACAM family, were not (Figure 1 and Table 1).

[00508] Higher levels of CEACAM5 mRNA were observed in high CEACAM5 expressers vs. moderate expressers (P = 0.0027) (Figure 1).

[00509] An association was found between the level of CEACAM5 mRNA in tumor cells and the IHC staining of CEACAM5 in tumor cells (Figure 2).

[00510] Furthermore, an enrichment of clinical responses was shown in patients treated with ADC who have a high level of CEACAM5 mRNA and a high expression of CEACAM5 protein, as measured by IHC (Figures 3 and 4).

[00511] As shown in Figure 5, illustrating the mRNA expression of CEACAM family members according to clinical responses to tusamitamab ravtansine, the median mRNA level of CEACAM5 was higher in the responder group compared to the non-responder group. This result shows an enrichment of clinical responses in patients treated with tusamitamab ravtansine according to CEACAM5 mRNA levels. This was not observed for other members of the CEACAM family, except for a slight trend for CEACAM3.

[00512] Clinical findings from the escalation and ex phases Petition 870250084766, dated 09 / 19 / 2025, pp. 119 / 149 111 / 115 dose-expanding clinical trials showed an enhancement of clinical responses in patients with high CEACAM5 protein expression (responders) treated with tusamitamab ravtansine versus patients treated with the ADC and with moderate CEACAM5 protein expression (non-responders).

[00513] Additional results from biomarker analysis support the association between the level of CEACAM5 mRNA expression and the level of CEACAM5 protein expression.

[00514] Since an association between the expression level of the CEACAM5 protein and the enhancement of clinical responses has been shown in patients treated with tusamitamab ravtansine, and since an association between the expression level of the CEACAM5 protein and the expression level of CEACAM5 mRNA has been shown, then the expression level of CEACAM5 mRNA has been shown to be a satisfactory biomarker of the enhancement of clinical responses and to select patients to be treated with an anti-CEACAM5 antibody-drug conjugate, such as tusamitamab ravtansine.

[00515] Furthermore, the level of CEACAM5 mRNA is a satisfactory biomarker for pre-screening cancer patients requiring cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated with a cytotoxic agent for a further screening step with immunohistochemical staining of CEACAM5 (IHC) and treating cancer patients requiring treatment with the ADC. [REFERENCES] 1. International Agency for Research on Cancer and WHO. Globocan: cancer today; mortality. October 12, 2020, 2021]; available at: https: / / gco.iarc.fr / today / online-analysis- pie?v=2020&mode=cancer&mode_population=continents&population= 900&populations=900&key=total&sex=0&cancer=39&type=1&statistic= Petition 870250084766, dated 09 / 19 / 2025, pp. 120 / 149 112 / 115 5&prevalence=0&population_group=0&ages_group%5B%5D=0&ages_ group%5B%5D=17&nb_items=15&group_cancer=1&include_nmsc=1& include_nmsc_other=1&half_pie=0&donut=0. 2. National Cancer Institute. Surveillance, Epidemiology, and End Results Program. Lung and bronchus: SEER 5-year ageadjusted incidence rates, 2015-2019. 12 de outubro de 2021, 2021]; Disponível em: https: / / seer.cancer.gov / explorer / application.html?site=47&data_type=1 &graph_type=10&compareBy=subtype&chk_subtype_47=47&chk_subt ype_612=612&chk_subtype_613=613&chk_subtype_611=611&chk_su btype_610=610&series=stage&chk_stage_101 = 101&chk_stage_104=1 04&chk_stage_105= 105&chk_stage_106= 106&chk_stage_107=107&s ex=1&race=1&age_range=1&advopt_precision=1. 3. National Cancer Institute. Surveillance, Epidemiology, and End Results Program. Lung and bronchus: SEER 5-year relative survival rates, 2012-2018. 12 de outubro de 2021, 2021]; disponível em: https: / / seer.cancer.gov / explorer / application.html?site=47&data_type=4 &graph_type=5&compareBy=subtype&chk_subtype_47=47&chk_subty pe_612=612&chk_subtype_613=613&chk_subtype_611=611&chk_sub type_610=610&series=stage&chk_stage_101 = 101&chk_stage_104=10 4&chk_stage_105=105&chk_stage_106=106&chk_stage_107=107&se x=1&race=1&age_range=1&advopt_precision=1. 4. Chu, QS. Targeting non-small cell lung cancer: driver mutation beyond epidermal growth factor mutation and anaplastic lymphoma kinase fusion. Ther Adv Med Oncol, 2020, 12: 1758835919895756. [PubMed ID: 32047535] 5. Rodak, O, Peris-Diaz, MD, Olbromski, M, et al. Current Landscape of Non-Small Cell Lung Cancer: Epidemiology, Histological Classification, Targeted Therapies, and Immunotherapy. Cancers (Ba Petição 870250084766, de 19 / 09 / 2025, pág. 121 / 149 113 / 115 sel), 2021, 13: 4705. [PubMed ID: 34572931] 6. Beauchemin, N and Arabzadeh, A. Carcinoembryonic antigen-related cell adhesion molecules (CEACAMs) in cancer progression and metastasis. Cancer Metastasis Rev, 2013, 32: 643-71. [PubMed ID: 23903773] 7. Decary, S, Berne, PF, Nicolazzi, C, et al. Preclinical Activity of SAR408701: A Novel Anti-CEACAM5-maytansinoid Antibodydrug Conjugate for the Treatment of CEACAM5-positive Epithelial Tumors. Clin Cancer Res, 2020, 26: 6589-6599. [PubMed ID: 33046521] 8. Decary, S, Berne, PF, Nicolazzi, C, et al. A novel antiCEACAM5 maytansinoid-antibody-drug conjugate for the treatment of colorectal, lung and gastric tumors. Cancer Research, 2015, 75: Abs 1688. [PubMed ID: WOS:000371578503178] 9. Blumenthal, RD, Hansen, HJ e Goldenberg, DM. Inhibition of adhesion, invasion, and metastasis by antibodies targeting CEACAM6 (NCA-90) and CEACAM5 (Carcinoembryonic Antigen). Cancer Res, 2005, 65: 8809-17. [PubMed ID: 16204051] 10. Zhang, X, Han, X, Zuo, P, et al. CEACAM5 stimulates the progression of non-small-cell lung cancer by promoting cell proliferation and migration. J Int Med Res, 2020, 48: 300060520959478. [PubMed ID: 32993395] 11. Hu, R, Huffman, KE, Chu, M, et al. Quantitative Secretomic Analysis Identifies Extracellular Protein Factors That Modulate the Metastatic Phenotype of Non-Small Cell Lung Cancer. J Proteome Res, 2016, 15: 477-86. [PubMed ID: 26736068] 12. Papadaki, MA, Messaritakis, I, Fiste, O, et al. Assessment of the Efficacy and Clinical Utility of Different Circulating Tumor Cell (CTC) Detection Assays in Patients with Chemotherapy-Naive Advanced or Metastatic Non-Small Cell Lung Cancer (NSCLC). Int J Mol Sci, 2021,22. [PubMed ID: 33477700] Petição 870250084766, de 19 / 09 / 2025, pág. 122 / 149 114 / 115 13. Desai, A, Abdayem, P, Adjei, AA, et al. Antibody-drug conjugates: A promising novel therapeutic approach in lung cancer. Lung Cancer, 2022, 163: 96-106. [PubMed ID: 34942494] 14. Pouzin, C, Gibiansky, L, Fagniez, N, et al. Integrated multiple analytes and semi-mechanistic population pharmacokinetic model of tusamitamab ravtansine, a DM4 anti-CEACAM5 antibodydrug conjugate. J Pharmacokinet Pharmacodyn, 2022, 49: 381-394. [PubMed ID: 35166967] 15. Gazzah, A, Bedard, PL, Hierro, C, et al. Safety, pharmacokinetics, and antitumor activity of the anti-CEACAM5-DM4 antibody-drug conjugate tusamitamab ravtansine (SAR408701) in patients with advanced solid tumors: first-in-human dose-escalation study. Ann Oncol, 2022, 33: 416-425. [PubMed ID: 35026412] 16. Gazzah, A, Ricordel, C, Cousin, S, et al. Efficacy and safety of the antibody-drug conjugate (ADC) SAR408701 in patients (pts) with non-squamous non-small cell lung cancer (NSQ NSCLC) expressing carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5). Journal of Clinical Oncology, 2020, 38: Abs 9505. [PubMed ID: WOS:000560368303410] 17. Ricordel, C, Barlesi, F, Cousin, S, et al. Safety and efficacy of tusamitamab ravtansine (SAR408701) in long-term treated patients with nonsquamous non-small cell lung cancer (NSQ NSCLC) expressing carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5). Journal of Clinical Oncology, 2022, 40: 9039-9039. [PubMed ID: 18. Sefrioui, D, Beaussire, L, Gillibert, A, et al. CEA, CA199, circulating DNA and circulating tumour cell kinetics in patients treated for metastatic colorectal cancer (mCRC). Br J Cancer, 2021, 125: 725-733. [PubMed ID: 34112948] 19. Gherman, A, Balacescu, L, Gheorghe-Cetean, S, et al. Petição 870250084766, de 19 / 09 / 2025, pág. 123 / 149 115 / 115 Current and New Predictors for Treatment Response in Metastatic Colorectal Cancer. The Role of Circulating miRNAs as Biomarkers. Int J Mol Sci, 2020, 21. [PubMed ID: 32197436] 20. Locker, GY, Hamilton, S, Harris, J, et al. ASCO 2006 update of recommendations for the use of tumor markers in gastrointestinal cancer. J Clin Oncol, 2006, 24: 5313-27. 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Claims

1. A method for selecting an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, characterized in that it comprises at least the steps of: (i) determining a gene expression level value of CEACAM5 in an isolated tumor sample obtained from said individual, (ii) comparing said determined value with a reference value, and (iii) selecting said individual for cancer treatment if the determined value is above the reference value.

2. Antibody-drug conjugate (ADC), characterized in that it comprises an anti-CEACAM5 antibody conjugated to a cytotoxic agent, for use in the treatment of cancer in an individual in need thereof, the use comprising: (i) determining a value of a CEACAM5 gene expression level in an isolated tumor sample obtained from said individual, (ii) comparing said determined value with a reference value and (iii) administering to said individual an effective amount of said ADC if said determined value is above the reference value.

3. Method for selecting an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, characterized by the fact that with Petition 870250084766, dated 09 / 19 / 2025, p.125 / 149 2 / 9 comprises at least the steps of: (i) determining a value of a CEACAM5 gene expression level in an isolated tumor sample obtained from said individual, (ii) comparing said value determined in step (i) with a reference value of a CEACAM5 gene expression level, (iii) selecting said individual for a CEACAM5 immunohistochemical staining (IHC) test if the value determined in step (i) is above the reference value of step (ii), (iv) determining an intensity of a CEACAM5 protein expression level with said CEACAM5 immunohistochemical staining (IHC) test in an isolated tumor sample obtained from said individual, (v) comparing said intensity determined in step (iv) with a reference intensity, and (vi) selecting said individual for cancer treatment if the intensity determined in step (iv) is above the reference intensity.

4. Antibody-drug conjugate (ADC), characterized in that it comprises an anti-CEACAM5 antibody conjugated to a cytotoxic agent, for use in the treatment of cancer in an individual in need thereof, the use comprising: (i) determining a value of a CEACAM5 gene expression level in an isolated tumor sample obtained from said individual, (ii) comparing said value determined in step (i) with a reference value of a CEACAM5 gene expression level, (iii) selecting said individual for a CEACAM5 immunohistochemistry (IHC) staining test if the value determined in Petition 870250084766, dated 19 / 09 / 2025, p.126 / 149 3 / 9 step (i) is above the reference value of step (ii), (iv) determine an intensity of expression level of the CEACAM5 protein with said CEACAM5 immunohistochemical staining (IHC) test in an isolated tumor sample obtained from said individual, (v) compare said intensity determined in step (iv) with a reference intensity, and (vi) administer to said individual an effective amount of said ADC if said intensity determined is above the reference intensity.

5. A method, as defined in either of claims 1 and 3, or an antibody-drug conjugate (ADC) for use, as defined in either of claims 2 and 4, characterized in that the value of the CEACAM5 gene expression level is a measure of a CEACAM5 gene transcript.

6. Antibody-drug conjugate (ADC) method or conjugate for use, as defined in claim 3, characterized in that the CEACAM5 gene transcript is an mRNA.

7. Method for selecting an individual in need of cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, characterized in that it comprises at least the steps of: (i) determining a quantile-normalized log2-transformed Transcripts Per Million Kilobases (TPM) value for CEACAM5 mRNA in a single tumor sample obtained from said individual, (ii) comparing said value with a reference value, and (iii) selecting said individual for cancer treatment if the determined value is above the reference value. Petition 870250084766, dated 19 / 09 / 2025, pp. 127 / 149 4 / 9 8. Antibody-drug conjugate (ADC), characterized in that it comprises an anti-CEACAM5 antibody conjugated to a cytotoxic agent, for use in the treatment of cancer in an individual in need thereof, the use comprising: (i) determining a quantile-normalized log2-transformed Transcripts Per Million Kilobases (TPM) value for CEACAM5 mRNA in a single tumor sample obtained from said individual, (ii) comparing said value with a reference value, and (iii) administering to said individual an effective amount of said ADC if the determined value is above the reference value.

9. Method for selecting an individual in need thereof for cancer treatment with an antibody-drug conjugate (ADC) comprising an anti-CEACAM5 antibody conjugated to a cytotoxic agent, characterized in that it comprises at least the steps of: (i) determining a quantile-normalized log2-transformed Transcripts Per Million Kilobases (TPM) value for CEACAM5 mRNA in a tumor isolate obtained from said individual, (ii) comparing said value determined in step (i) with a reference value, (iii) selecting said individual for a CEACAM5 immunohistochemistry (IHC) staining test if the value determined in step (i) is above the reference value of step (ii), (iv) determining the intensity of a CEACAM5 protein expression level with said CEACAM5 immunohistochemistry (IHC) staining test in a tumor isolate obtained from said individual, Petition 870250084766, of 19 / 09 / 2025, page.128 / 149 5 / 9 (v) compare said intensity determined in step (iv) with a reference intensity, and (vi) select said individual for cancer treatment if the determined intensity is above the reference intensity.

10. Antibody-drug conjugate (ADC), characterized in that it comprises an anti-CEACAM5 antibody conjugated to a cytotoxic agent, for use in the treatment of cancer in an individual in need thereof, the use comprising: (i) determining a quantile-normalized Transcripts Per Million Kilobases (TPM) value transformed by log2 for CEACAM5 mRNA in a single tumor sample obtained from said individual, (ii) comparing said value determined in step (i) with a reference value, (iii) selecting said individual for a CEACAM5 immunohistochemical staining (IHC) test if the value determined in step (i) is above the reference value of step (ii), (iv) determining the intensity of a CEACAM5 protein expression level with said CEACAM5 immunohistochemical staining (IHC) test in a single tumor sample obtained from said individual,(v) compare the said intensity determined in step (iv) with a reference intensity, and (vi) administer to said individual an effective amount of said ADC if the determined intensity is above the reference intensity.

11. Method, as defined in any one of claims 1, 3, 5 to 7 and 9, or the antibody-drug conjugate (ADC) Petition 870250084766, dated 09 / 19 / 2025, p. 129 / 149 6 / 9 for use, as defined in any one of claims 2, 4 to 6, 8 and 10, characterized in that the anti-CEACAM5 antibody comprises an HCDR1 having the amino acid sequence of SEQ ID NO: 1, an HCDR2 having the amino acid sequence of SEQ ID NO: 2, an HCDR3 having the amino acid sequence of SEQ ID NO: 3, an LCDR1 having the amino acid sequence of SEQ ID NO: 4, an LCDR2 having the amino acid sequence NTR and an LCDR3 having the amino acid sequence of SEQ ID NO:

5.

12. A method, as defined in any one of claims 1, 3, 5 to 7, 9 and 11, or the antibody-drug conjugate (ADC) for use, as defined in any one of claims 2, 4 to 6, 8 and 10 and 11, characterized in that the cytotoxic agent is maytansinoid or a maytansinoid analogue.

13. Method or use, as defined in any one of claims 7, 9, 11 and 12, or antibody-drug conjugate (ADC) for use, as defined in any one of claims 8 and 10 to 12, characterized in that the reference value is at least about 7 to about 13.

14. Method, as defined in any one of claims 7, 9 and 11 to 13, or antibody-drug conjugate (ADC) for use, as defined in any one of claims 8 and 10 to 13, characterized in that the reference value is at least about 7, or is at least about 8, or at least about 9, or at least about 10, or at least about 11, or at least about 12, or at least about 13.

15. Method, as defined in any one of claims 7, 9 and 11 to 14, or antibody-drug conjugate (ADC) for use, as defined in any one of claims 8 and 10 to 13, characterized in that quantile normalization is obtained by (i) classifying sample transcripts by expression level, (ii) calculating an average value for genes occupying the same class and (iii) replacing the values ​​of all genes occupying said class by that average value.

16. Method, as defined in any one of claims 7, 9 and 11 to 15, or antibody-drug conjugate (ADC) for use, as defined in any one of claims 8 and 10 to 15, characterized in that the expression level of transcripts is measured in Fragments Per Million Kilobases (FPKM) before being converted to TPM.

17. Antibody-drug conjugate (ADC) method or conjugate for use according to claim 16, characterized in that Fragments per Million Kilobases (FPKM) are obtained by counting the total transcripts in said sample, dividing the transcript counts obtained by 1,000,000 and dividing the values ​​obtained by the gene length, in kilobases.

18. Antibody-drug conjugate (ADC) method or conjugate for use according to any one of claims 1 to 17, characterized in that the cancer is selected from the group consisting of hepatocellular carcinoma, colorectal cancer, gastric cancer, gastroesophageal junction (GEJ) adenocarcinoma, esophageal cancer, lung cancer, cervical 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.

19. Antibody-drug conjugate (ADC) method or conjugate for use, as defined in any one of claims 1 to 18, characterized in that the ADC is tusamitamab ravtansine.

20. Antibody-drug conjugate (ADC) method or conjugate for use, as defined in any of claims 1 to 19, characterized in that the ADC is administered at a dose of > 80 mg. Petition 870250084766, dated 09 / 19 / 2025, p. 131 / 149 8 / 9 mg / m2, in particular 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 in particular at a dose of 80, 100, 120, 150, 170, 180 or 210 mg / m2, relative to the body surface area of ​​said individual, once every two weeks, or the ADC is administered at a dose of > 80 mg / m2, relative to the body surface area of ​​said individual approximately once every three weeks.

21. Antibody-drug conjugate (ADC) method or conjugate for use, as defined in any one of claims 2, 4 to 6 and 8, 10 to 16, characterized in that it further comprises administering to the individual an effective amount of at least one additional agent effective for treating cancer.

22. Antibody-drug conjugate (ADC) method or conjugate for use according to claim 21, characterized in that the additional agent is selected from the group consisting of an immune checkpoint inhibitor (ICI), in particular an anti-PD-1 antibody or an anti-PD-L1 antibody, a platinum-based chemotherapy, pemetrexed, anti-VEGFR2, FOLFOX, FOLFIRI, TAS102, anti-EGFR and any combination thereof.

23. Antibody-drug conjugate (ADC) method or method for use according to claim 22, characterized in that the anti-PD-1 antibody is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, sintilimab, dostarlimab and tislelizumab or in that the anti-PD-L1 antibody is selected from the group consisting of atezolizumab, avelumab and durvalumab.

24. Antibody-drug conjugate (ADC) method or method for use according to claim 23, characterized in that it comprises administering to the individual an effective amount of tusamitamab ravtansine and pembrolizumab. Petition 870250084766, dated 09 / 19 / 2025, pp. 132 / 149 9 / 9 25. A method or antibody-drug conjugate (ADC) for use according to claim 22, characterized in that it further comprises administering to the individual an effective amount of a platinum-based chemotherapy.

26. Antibody-drug conjugate (ADC) method or conjugate for use according to claim 25, characterized in that the platinum-based chemotherapy is selected from cisplatin and carboplatin.

27. Antibody-drug conjugate (ADC) method or conjugate for use, according to any one of claims 25 to 26, characterized in that it further comprises administering an effective amount of pemetrexed to the individual.

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