Combination of antibodies or antigen-binding fragments thereof for cancer treatment

Abstract

The present invention relates to the use of a combination of antibodies or antigen-binding fragments thereof for the treatment of cancer in a human subject in need thereof, the combination comprising: at least a first antibody or antigen-binding fragment thereof, the first antibody or antigen-binding fragment thereof specifically binding to FASN; at least a second antibody or antigen binding fragment thereof, said second antibody or antigen binding fragment thereof specifically binding to CD99; and at least a third antibody or antigen-binding fragment thereof, said third antibody or antigen-binding fragment thereof specifically binding to an antigen selected from the group consisting of GPI, SLC3A2, GART and CKAP4, in particular the group consisting of GPI and GART. The invention further relates to pharmaceutical compositions comprising a combination of such antibodies or antigen-binding fragments thereof and a pharmaceutically acceptable carrier for the treatment of cancer in a human subject in need thereof.

Description

Technical Field

[0001] This invention relates to the field of cancer treatment. In particular, this invention relates to the use of polyclonal antibodies or combinations of antibodies against specific antigens and their application in cancer treatment. Background Technology

[0002] Cancer is the second leading cause of death worldwide, causing an estimated 9.6 million deaths in 2018. Globally, approximately one in six deaths are caused by cancer. Inadequate or insufficiently aggressive treatment is common in this field. Therefore, the development of novel and improved therapies for cancer is a recognized need.

[0003] In cancer, PTCL (or peripheral T-cell lymphoma) constitutes a diverse group of lymphoproliferative tumors originating from mature retrothymic T cells. This group of tumors exhibits significant heterogeneity, with most cases displaying aggressive invasiveness. PTCL can be classified into four main groups, encompassing over 30 subtypes, including peripheral T-cell lymphoma nonspecific, angioimmunoblastic T-cell lymphoma, extranodal NK / T-cell lymphoma (nasal type), ALK+ systemic anaplastic large T-cell lymphoma, ALK- systemic anaplastic large T-cell lymphoma, mycosis fungoides, and Sézary syndrome, among others. This complex pathological classification system reflects the extensive variability within this disease group.

[0004] Geographically, the incidence and distribution of PTCL subtypes vary. Asia has a slightly higher incidence compared to Europe and the United States. Notably, in China, PTCL accounts for approximately 25% to 35% of non-Hodgkin lymphoma (NHL), significantly higher than the 10% to 15% observed in Europe and the United States.

[0005] Immunotherapy has brought real hope to the treatment of many types of cancer, especially those that are currently incurable by conventional therapies. This treatment primarily involves administering monoclonal antibodies that target tumor cells or activators or inhibitors of the immune response checkpoints against cancer. In addition to these immunomodulatory monoclonal antibodies (mAbs), antibody-drug conjugates (ADCs), chimeric antigen receptor T cells (CAR-T), and more recently, bispecific antibodies are being actively explored and successfully incorporated into the arsenal of innovative cancer therapies. However, for many cancers, effective treatments remain lacking, particularly those leading to long-term cancer-free survival and reducing the risk of metastasis and recurrence.

[0006] Antibodies targeting tumors (passive immunotherapy) can exert their effects through two complementary mechanisms:

[0007] - Through complement-dependent cytotoxicity (CDC), complement-dependent cytotoxicity (ADCC) from killer cells, or complement-dependent cytotoxicity (ADCP) from phagocytes.

[0008] - By inducing an adaptive immune response. It has indeed been demonstrated that opsonization of the target and local production of complement molecules (C3a, C5a) (see Strainic et al., Immunity, 2008 Mar;28(3):425-35) activate T lymphocyte co-stimulation and enhance their survival. Therefore, passive administration of antitumor antibodies can, as a first step, generate the expulsion of factors from complement, which in a second step can promote a T cell response.

[0009] More specifically regarding PTCL, several monoclonal antibodies are currently being investigated for the treatment of these cancers, including many novel drugs being investigated in clinical trials for the treatment of PTCL, including aisertib, bendamustine (Treanda), bortezomib (Velcade), lenalidomide (Revlimid), nivolumab (Opdivo), panobinostat (Farydak), and pembrolizumab (Keytruda).

[0010] However, 40% to 85% of patients are resistant to monoclonal antibody-based therapies. Escape mechanisms include immune selection mechanisms (i.e., the ability of tumor cells to lose the ability to recognize antigens by the immune system) and immune subversion mechanisms (the induction of specific tolerance). The emergence of tumor variants with lower immunogenicity can be particularly detrimental in monoclonal antibody-based therapies (which specifically target unique epitopes).

[0011] More specifically, the prognosis for patients with PTCL is not optimistic. The overall response rate to initial treatment with the CHOP regimen (cyclophosphamide, doxorubicin, vincristine, and prednisone) or CHOP-based regimens is relatively low, and PTCL tends to relapse, leading to a poor prognosis. Despite recent significant advances in understanding PTCL, including new laboratory diagnostic methods and new treatment approaches, the prognosis for most PTCL cases remains worse than that for aggressive B-cell lymphomas. Except for ALK+ systemic anaplastic large T-cell lymphoma (ALCL), the 5-year survival rate for other PTCL subtypes ranges from 14% to 32% (Vose J et al., International peripheral T-cell and natural killer / T-cell lymphoma study: pathologyfindings and clinical outcomes. J Clin Oncol. 2008 Sep 1; 26(25): 4124-4130).

[0012] Therefore, treatment based on combinations of antibodies targeting different antigens on tumor cells can allow for minimization of escape mechanisms.

[0013] In particular, the polyclonal and multi-targeting nature of antibody combinations can avoid the shortcomings encountered by monoclonal antibody multi-isotope or single-target methods.

[0014] However, despite Richet's observation of the potential efficacy of antibody combinations over 100 years ago, combinations of antibodies, such as animal-derived polyclonal antibodies, are rarely used for cancer treatment, particularly due to the high risk of toxicity in patients. This toxicity is primarily associated with the expression of Neu5GC and α-1,3-galactose carbohydrates on animal immunoglobulins, which elicit potent anti-Neu5GC and anti-α-1,3-galactose immune responses in humans, and are associated with allergies, serum sickness, and the formation of immune complexes. Combinations of monoclonal antibodies have been considered to mimic the polyclonal humoral immune responses used in cancer treatment. However, the clinical efficacy of monoclonal antibodies as monotherapy does not predict their safety and clinical efficacy in combination (Berlin et al., Investigational News Drugs (2022) 40:586-595). The main difficulty with this combination lies in the association of antibodies with different PKs (which makes controlling toxicity more difficult), the stability of the homologous product, and manufacturing (see, for example, Larbouret et al., Cancers (Basel). 15 Sep 2021;13(18):4620).

[0015] Therefore, there is still a need in this field for novel anticancer therapeutics, and especially for novel therapeutics that are effective against a variety of cancers.

[0016] Therefore, there is still a need in the field for novel anticancer therapeutics, and in particular, novel therapeutics that are effective against multiple PTCL subtypes.

[0017] There is still a need in the field for effective and improved therapies for cancer treatment, with reduced or no adverse reactions, and especially very low interaction with PBMCs.

[0018] There is still a need in this field to provide novel anticancer therapeutics that have anticancer activity against both solid and liquid cancers.

[0019] There is still a need in the field to provide novel anticancer therapies that are effective against PTCL subtypes resistant to standard care CHOP or CHOEP (CHOP + etoposide).

[0020] There is still a need in the art to provide novel anticancer therapeutics that can activate complement and apoptosis mechanisms to kill a variety of cancers, and novel anticancer therapeutics that specifically activate both intrinsic and extrinsic apoptosis pathways, and particularly by inducing both caspase 8 and caspase 9.

[0021] There is still a need in the art to provide novel anticancer therapeutic agents capable of providing antitumor activity selected from the group consisting of complement-dependent cytotoxicity (CDC), complement-dependent cytotoxicity (ADCC) from killer cells, apoptotic activity and / or complement-dependent cytotoxicity (ADCP) from phagocytes, and particularly antitumor activity consisting of CDC, ADCC, apoptotic activity and ADCP.

[0022] The purpose of this invention is to meet the above-mentioned needs. Summary of the Invention

[0023] This invention specifically relates to the following:

[0024] Item 1 : A combination of antibodies or antigen-binding fragments thereof, the combination for the treatment of cancer in human subjects of need, the combination comprising:

[0025] - At least a first antibody or its antigen-binding fragment thereof, wherein the first antibody or its antigen-binding fragment specifically binds to the FASN (FasN fragment);

[0026] - At least a second antibody or its antigen-binding fragment, wherein the second antibody or its antigen-binding fragment specifically binds to CD99 (differentiation cluster 99); and

[0027] - At least a third antibody or its antigen-binding fragment, said third antibody or its antigen-binding fragment specifically binding to an antigen selected from the group consisting of GPI (glucose-6-phosphate isomerase), SLC3A2 (solute carrier family 3 member 2), GART (glycamide ribonucleotide formyltransferase) and CKAP4 (cytoskeleton-associated protein 4), particularly the group consisting of GPI and GART.

[0028] Item 2 The combination used according to the purpose described in item 1 further comprises at least a fourth antibody or an antigen-binding fragment thereof, the fourth antibody or an antigen-binding fragment thereof specifically binding to an antigen selected from the group consisting of GPI, SLC3A2, GART and CKAP4, particularly the group consisting of GPI and GART, the fourth antibody or an antigen-binding fragment thereof being different from the third antibody or an antigen-binding fragment thereof.

[0029] Item 3 The combination used according to the purpose described in item 1 or item 2, wherein the combination comprises at least:

[0030] -At least a first antibody or its antigen-binding fragment thereof, wherein the first antibody or its antigen-binding fragment specifically binds to FASN;

[0031] -At least a second antibody or its antigen-binding fragment thereof, wherein the second antibody or its antigen-binding fragment specifically binds to CD99;

[0032] - At least a third antibody or its antigen-binding fragment, said third antibody or its antigen-binding fragment specifically binding to GPI; and

[0033] - At least a fourth antibody or its antigen-binding fragment, said fourth antibody or its antigen-binding fragment specifically binds to GART.

[0034] Item 4 The combination used according to any one of the uses described in items 1 to 3, wherein the antigen-binding fragment of the antibody present in the combination is independently Fv, Fab, F(ab')2, Fab', dsFv, (dsFv)2, scFv, sc(Fv)2 or a double-chain antibody.

[0035] Item 5 The combination of uses according to any one of items 1 to 4, wherein the cancer is selected from the group consisting of: myeloma; melanoma; skin cancer; breast cancer; brain tumors, bladder cancer, cervical cancer, prostate cancer; primary and metastatic colorectal cancer, especially colon cancer; mesothelioma; lung cancer, especially non-small cell lung cancer; liver cancer, especially hepatocellular carcinoma or bile duct cancer; primary and metastatic pancreatic cancer; kidney cancer; soft tissue sarcoma; thyroid cancer; lymphoma, including Hodgkin lymphoma and non-Hodgkin lymphoma, especially B-cell lymphoma or T-cell lymphoma, more particularly T-cell lymphoma; gastric cancer; head and neck cancer; ovarian cancer; sarcoma; acute or chronic leukemia, especially T-cell leukemia or myeloid leukemia; osteosarcoma; anal cancer; testicular cancer; uterine cancer; thyroid cancer; central nervous system cancers; gastrointestinal stromal cancer; epidermal cancer; and esophageal cancer.

[0036] Item 6 The combination of uses according to any one of items 1 through 5, wherein the cancer is:

[0037] - Lymphoma, especially those selected from the group consisting of the following:

[0038] (i) Non-Hodgkin lymphoma selected from the group consisting of the following:

[0039] - B-cell lymphomas, particularly those selected from the group consisting of: diffuse large B-cell lymphoma, follicular lymphoma, small lymphocytic lymphoma (or chronic lymphocytic leukemia), mantle cell lymphoma, marginal zone B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma (or Waldenstrom macroglobulinaemia), hairy cell leukemia, and primary central nervous system lymphoma; and

[0040] -T-cell lymphomas, particularly those selected from the group consisting of: precursor T-lymphoblastic lymphoma (or precursor T-lymphoblastic leukemia), peripheral T-cell lymphoma, and cutaneous T-cell lymphoma; and

[0041] (ii) Hodgkin lymphoma selected from the following groups: tuberous sclerosis Hodgkin lymphoma, mixed cellularity Hodgkin lymphoma, lymphocytic depletion Hodgkin lymphoma, and lymphocyte-rich Hodgkin lymphoma; or

[0042] - Leukemia, particularly those selected from the following groups: acute myeloid leukemia (AML), chronic myeloid leukemia (CML), and histiocytic leukemia.

[0043] Item 7 The combination of uses according to any one of items 1 through 6, wherein the cancer is selected from the group consisting of:

[0044] -T-cell acute lymphoblastic leukemia (T-ALL);

[0045] - Peripheral T-cell lymphoma (PTCL), especially PTCL selected from the following groups: PTCL nonspecific type (PTCL-NOS), enteropathy-associated T-cell lymphoma (EATL), monomorphic epithelial-friendly intestinal T-cell lymphoma (MEITL), anaplastic large cell lymphoma (ALCL) and angioimmunoblastic T-cell lymphoma (AITL), extranodal NK / T-cell lymphoma nasal type (ENKL), hepatosplenic γδ T-cell lymphoma (HSGDTCL), intestinal T-cell lymphoma (ITCL), mycosis fungoides and Cezari syndrome;

[0046] - Cutaneous T-cell lymphoma (CTCL); and

[0047] -T-cell lymphoblastic lymphoma (T-LBL).

[0048] Item 8The combination used according to any one of items 1 to 7, wherein the cancer expresses at least one antigen selected from the group consisting of FASN, CD99, GPI, SLC3A2, GART and CKAP4, particularly the group consisting of GPI and GART.

[0049] Item 9 The combination used according to any one of items 1 to 8, wherein at least one antibody in the combination, and in particular all antibodies in the combination, lacks at least one antigenic determinant selected from (i) N-hydroxyacetylneuraminic acid (Neu5Gc) and (ii) α-1,3-galactose, and in particular lacks two antigenic determinants: N-hydroxyacetylneuraminic acid (Neu5Gc) and α-1,3-galactose.

[0050] Item 10 : A combination used according to any one of the uses described in items 1 to 9, wherein the combination is a combination of monoclonal antibodies or a polyclonal antibody composition.

[0051] Item 11 The combination used according to the purpose described in item 10, wherein the polyclonal antibody composition can be obtained by immunizing animals with CD3+, CD8+, TCR γ / δ- and CD34- human tumor T cells.

[0052] Item 12 The combination used according to the purpose described in item 11, wherein the animal is a pig, and particularly a pig lacking at least one gene selected from the group comprising (i) a gene encoding a functional cytidine-5'-monophosphate N-acetylneuraminic acid hydrolase (CMAH) and (ii) a gene encoding a functional α-(1,3)-galactosyltransferase, and more particularly a pig lacking both (i) a gene encoding a functional cytidine-5'-monophosphate N-acetylneuraminic acid hydrolase (CMAH) and (ii) a gene encoding a functional α-(1,3)-galactosyltransferase.

[0053] Item 13 : A combination used according to any one of the uses described in items 1 to 12, wherein the combination includes a pharmaceutical composition, the pharmaceutical composition further comprising a pharmaceutically acceptable carrier.

[0054] Item 14 The combination used according to the purpose described in item 13, wherein the pharmaceutical composition further comprises at least one additional anticancer drug, said additional anticancer drug being different from the antibody in the combination of antibodies defined in any one of items 1 to 4 and 8 to 11.

[0055] Item 15 The combination used according to the purpose described in item 14, wherein the at least one additional anticancer drug is selected from the group consisting of monoclonal antibodies, particularly from the group consisting of the following monoclonal antibodies: anti-CD19, anti-CD20, anti-CD30, anti-CD137, anti-CTLA4, anti-TIM-3, anti-B7-H3, anti-CD123, anti-CD134, anti-CD154, anti-LAG-3, anti-CD227, anti-BTNA3, anti-CD39, anti-CD73, anti-CD115, anti-CD47, anti-SIRPα, anti-SIRPγ, anti-CD28, anti-NCR, anti-NKp46, anti-NKp30, anti-NKp44, anti-NKG2D, anti-PD1, anti-PDL1, mogamulizumab, obinutuzumab, polatuzumab vedotin, yttrium Y90-tiemomumab. Y90-ibritumomab tiuxetan), mosunetuzumab, and anti-DNAM-1 monoclonal antibody.

[0056] Item 16 The combination used according to any one of items 13 to 15, wherein the pharmaceutical composition further comprises at least one chemotherapy treatment, particularly chemotherapy treatment selected from the group consisting of: a chemotherapy regimen consisting of cyclophosphamide, hydroxydaunorubicin, vincristine, and prednisone (CHOP); a chemotherapy regimen consisting of cyclophosphamide, hydroxydaunorubicin, vincristine, etoposide, and prednisone (CHOEP); an HDAC inhibitor; ibrutinib, acalabrutinib, zanubrutinib, copanlisib, and venetoclax.

[0057] Item 17 The use of a combination according to any one of items 1 to 16, the combination being used to provide antitumor activity selected from the group consisting of complement-dependent cytotoxicity (CDC), complement-dependent cytotoxicity (ADCC) from killer cells, apoptotic activity and / or complement-dependent cytotoxicity (ADCP) from phagocytes, and particularly for providing antitumor activity consisting of complement-dependent cytotoxicity (CDC), complement-dependent cytotoxicity (ADCC) from killer cells, apoptotic activity and complement-dependent cytotoxicity (ADCP) from phagocytes. Attached Figure Description

[0058] Figure 1 The figure describes the cytotoxic activity of antibody combinations against different cell lines. The percentage (%) of cytotoxicity of PA1 (at a concentration of 100 µg / mL) against the following cell lines (from left to right): PBMC, HPB-ALL, Jurkat, T1301, HUT78, and OCI-LY-12.

[0059] Figure 2 This figure describes the antitumor activity of the antibody combination against the CHOP-resistant PTCL-NOS cell line. The percentage of cell death (%) is shown at different PA1 concentrations (µg / mL).

[0060] Figure 3 The figure describes the cytotoxic activity of antibody combinations and alemtuzumab (an anti-CD52 monoclonal antibody) against different cell lines. The figure shows the percentage (%) of cytotoxicity of PA1 (100 µg / mL, left column in each group) and alemtuzumab (100 µg / mL, right column in each group) against the following cell lines (from left to right): PBMC, HPB-ALL, Jurkat, and T1301.

[0061] Figure 4 This figure describes the apoptotic activity of antibody combinations against different cell lines. The percentage (%) of apoptosis in the following cell lines at different PA1 concentrations (µg / mL) is shown: PBMC (● curve), Jurkat (■ curve), T1301 (…). curve), HPB-ALL ( (curve) and OCI-LY-12 (◆curve).

[0062] Figure 5 The figure describes the combination of antibodies and the apoptotic activity of alemtuzumab against different cell lines. The percentage (%) of apoptotic activity of PA1 (30 µg / mL, left column in each group) and alemtuzumab (30 µg / mL, right column in each group) against the following cell lines (from left to right): HPB-ALL, T1301, and Jurkat.

[0063] Figure 6 The figure describes the ADCC activity of the antibody combination against the HPB-ALL cell line. The percentage of cell death (%) is shown at PA1 concentrations of 10 µg / mL (left column), 30 µg / mL (middle column), or 100 µg / mL (right column).

[0064] Figure 7 The figure describes the ADCP activity of the antibody combination against the HPB-ALL cell line. The percentage of cell death (%) is shown at PA1 concentrations of 10 µg / mL (left column), 30 µg / mL (middle column), or 100 µg / mL (right column).

[0065] Figure 8 This figure describes the activation of caspase 8 and caspase 9 in different cell lines by combinations of antibodies. The figure shows the percentage (%) of caspase 8 (two columns on the left of each chart) or caspase 9 (two columns on the right of each chart) positive cells in each cell line with or without PA1 (CT column). Paired t-tests were performed. p<0.05; p<0.01; p < 0.001.

[0066] Figure 9 This figure describes the evolution of tumor size in a T1301 xenograft mouse model in the presence of a combination of antibodies. The figure shows the mean tumor size (mm) in the presence of PA1 (■ curve) or in the absence of treatment (● curve). 3 The variation of ) over time (number of days). N=10 for each group. p<0.05 – Mann-Whitney test.

[0067] Figure 10 This figure describes the tumor size in a T1301 xenograft mouse model 35 days after combination therapy with antibodies. The figure represents the average tumor size (mm) at day 35 with or without PA1 (right column). 3 ). N=10 for each group. p<0.05 – Mann-Whitney test.

[0068] Figure 11 This figure depicts the evolution of tumor size in a Jurkat xenograft mouse model in the presence of a combination of antibodies. The figure shows the effect of PA1 (…). The mean tumor size (mm3) over time (days) is shown in the case of a curve (■ curve) or in the case of no treatment (■ curve). N=10 per group. p<0.05 – Mann-Whitney test.

[0069] Figure 12 This figure depicts tumor size in a Jurkat xenograft mouse model 40 days after combination antibody treatment. The figure represents the mean tumor size (mm) at day 40 with or without PA1 (right column). 3 ). N=10 for each group. p<0.05 – Mann-Whitney test.

[0070] Figure 13 The figure describes the CDC of antibody combinations against solid cancers. The figure shows the percentage (%) of cytotoxicity to the following cell lines at different PA1 concentrations (µg / mL): A549 (●), HepG2 (■), HCT116 (…). ), MDA-MB231 ( ), LNCAP (◆) and SKMEL30 (○).

[0071] Figure 14 This figure describes the overall cytotoxicity of the antibody combination against solid cancers. The figure represents the percentage (%) of cytotoxicity against the following cell lines at different PA1 concentrations (µg / mL): A549 (●), HCT116 ( LNCAP ) and SKMEL30 (◆).

[0072] Figure 15 This figure describes the binding of antibody combinations to different protein targets. The graph shows the Z-scores of PA1 for the following targets (from left to right): FasN, GPI, CD99, GART, SLC3A2, and CKAP4.

[0073] Figure 16 The following groups are described in terms of tumor size (mm). 3 Changes in treatment duration (number of days): Control group (●), PA1 group (■), and CHOP group ( The arrows below the figure indicate the days when PA1 treatment was administered (One-way ANOVA – Post-hoc Fischer test). p<0.05, p<0.01).

[0074] Figure 17 This describes the mean tumor volume (mm²) for each group at D28 (day 28 after treatment initiation). 3 From left to right, the groups are: control group (dark gray), CHOP group (light gray), and PA1 group (gray) (one-way ANOVA – post-hoc Fischer test). p<0.05).

[0075] Figure 18 The percentage of cell death (%) in different cell lines based on PA1 concentration (µg / ml) is described: HPB-ALL (●), T-1301 (■), Jurkat ( HUT78 ) and KARPASS 299 (◆). Detailed Implementation

[0076] 1. Definition

[0077] Some definitions are described below. Such definitions are intended to cover syntactic equivalences.

[0078] Unless otherwise defined herein, scientific and technical terms used in connection with this invention shall have the meanings commonly understood by one of ordinary skill in the art. For example, *The Concise Dictionary of Biomedicine and Molecular Biology*, Juo, Pei-Show, 2nd edition, 2002, CRC Press; *The Dictionary of Cell and Molecular Biology*, 3rd edition, 1999, Academic Press; and *The Oxford Dictionary of Biochemistry and Molecular Biology*, revised edition, 2000, Oxford University Press, can provide a general dictionary for those skilled in the art of the many terms used in this disclosure. In case of conflict, this specification (including definitions) shall prevail. Exemplary methods and materials are described below, but similar or equivalent methods and materials may also be used in the practice or testing of this invention. Units, prefixes, and symbols are expressed in their International System of Units (SI) recognized form. The headings provided herein are not intended to limit the various aspects of this disclosure.

[0079] All publications and other references mentioned in this article are incorporated herein by reference in their entirety.

[0080] It should be noted that the terms "an" or "a" entity refer to one or more of the entity; for example, "an antibody" should be understood to mean one or more antibodies. Therefore, the terms "an" (or "a"), "one or more," and "at least one" are used interchangeably in this document.

[0081] Throughout this specification and embodiments, the terms “having” and “comprising,” or variations such as “owning,” “having,” “including,” or “containing,” are to be understood as implying inclusion of the stated integers or groups of integers, but not excluding any other integers or groups of integers. The terms “consisting of” imply inclusion of the stated element (such as a composition of matter or a method step), but not excluding any other element. The term “composed of” imply inclusion of the stated element, excluding any additional element. The term “substantially composed of” imply inclusion of the stated element, as well as possible other elements, provided that such other elements do not materially affect the essential characteristics of this disclosure. It should be understood that different embodiments of this disclosure use the term “comprising” or its equivalents to cover embodiments in which the term is replaced with “comprising only,” “composed of,” or “substantially composed of.”

[0082] It should be understood that aspects described in this article using the language of "include" are also provided with similar aspects described using "consisting of" and / or "substantially composed of".

[0083] Furthermore, the use of “and / or” herein should be considered as a specific disclosure of two particular features or components, regardless of the presence of the other. Therefore, the term “and / or” as used herein in phrases such as “A and / or B” is intended to include “A and B”, “A or B”, “A” (alone), and “B” (alone). Similarly, the term “and / or” as used in phrases such as “A, B, and / or C” is intended to cover 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).

[0084] "Pharmaceutical-acceptable" or "pharmaceutical-admissible" means a molecular entity or composition that, when administered to mammals (especially humans) where appropriate, will not produce adverse reactions, allergic reactions, or other adverse effects. Pharmaceutically acceptable carriers or excipients refer to any type of non-toxic solid, semi-solid, or liquid filler, diluent, encapsulating material, or formulation adjuvant.

[0085] As used herein, “pharmaceutically acceptable carriers” include any and all physiologically compatible solvents, dispersion media, coatings, antimicrobial agents, and antifungal agents. Examples of suitable carriers, diluents, and / or excipients include one or more of the following: water, amino acids, saline, phosphate-buffered saline, phosphate buffer, acetate, citrate, succinate; amino acids and derivatives such as histidine, arginine, glycine, proline, glycylglycine; inorganic salts, NaCl, calcium chloride; sugars or polyols such as glucose, glycerol, ethanol, sucrose, trehalose, mannitol; surfactants such as polysorbate 80, polysorbate 20, poloxamer 188; and combinations thereof. In many cases, it is preferred to include isotonic agents, such as sugars, polyols, or sodium chloride, and the formulation may also contain antioxidants (such as tryptamine) and stabilizers (such as Tween 20). Suitable excipients and pharmaceutical formulation requirements are described in "Remington: The Science & Practice of Pharmacy," a reference work in the field.

[0086] As used herein, the term “antibody” has the same meaning and is used equivalently in this specification. As used herein, the term “antibody” refers to isolated or recombinant immunoglobulin molecules and the immunoactive portion of immunoglobulin molecules, i.e., molecules containing antigen-binding sites that specifically bind to antigens such as proteins FASN, CD99, SLC3A2, GART, or CKAP4. Therefore, the term antibody covers not only complete antibody molecules but also antibody fragments and variants (including derivatives) of antibodies and antibody fragments. In natural antibodies, two heavy chains are linked to each other by disulfide bonds, and each heavy chain is linked to a light chain by a disulfide bond. There are two types of light chains: λ (1) and κ (k). There are five main classes (or isotypes) of heavy chains that determine the functional activity of the antibody molecule: IgM, IgD, IgG, IgA, and IgE. Each chain contains different sequence domains. The light chain includes two domains: a variable domain (VL) and a constant domain (CL). The heavy chain comprises four domains: a variable domain (VH) and three constant domains (CH1, CH2, and CH3, collectively referred to as CH). Both the light chain variable region (VL) and the heavy chain variable region (VH) determine antigen binding recognition and specificity. The light chain constant region domain (CL) and the heavy chain constant region domain (CH) confer important biological properties such as antibody chain association, secretion, transplacental movement, complement binding, and binding to the Fc receptor (FcR). The Fv fragment is the N-terminal portion of the Fab fragment of an immunoglobulin and consists of a variable portion of one light chain and one heavy chain. Antibody specificity lies in the structural complementarity between the antibody binding site and the antigenic determinant. The antibody binding site is primarily composed of residues from the hypervariable region or complementarity-determining region (CDR). Occasionally, residues from the non-hypervariable region or framework region (FR) can participate in the antibody binding site or influence the overall domain structure, thus affecting the binding site. Complementarity-determining regions (CDRs) are amino acid sequences that collectively define the binding affinity and specificity of the native Fv region of the immunoglobulin binding site. Each of the light and heavy chains of immunoglobulins has three CDRs, named CDR1-L, CDR2-L, L-CDR3-L and CDR1-H, CDR2-H, CDR3-H, respectively. Therefore, the antigen-binding site (typically comprising six CDRs) contains a set of CDRs from each heavy chain V region and light chain V region. Frame regions (FRs) are amino acid sequences situated between the CDRs.

[0087] The term "antibody" includes, but is not limited to, monoclonal antibodies, polyclonal antibodies, human antibodies, humanized antibodies, camel antibodies, and chimeric antibodies. Antibodies can be any isotype / class (e.g., IgG, IgE, IgM, IgD, IgA, and IgY) or subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2).

[0088] In the context of this disclosure, the term "antibody" specifically includes antibodies that bind to FASN proteins (also known as anti-FASN antibodies), and / or antibodies that bind to CD99 proteins (also known as anti-CD99 antibodies), and / or antibodies that bind to GPI proteins (also known as anti-GPI antibodies), and / or antibodies that bind to SLC3A2 proteins (also known as anti-SLC3A2 antibodies), and / or antibodies that bind to GART proteins (also known as anti-GART antibodies), and / or antibodies that bind to CKAP4 proteins (also known as anti-CKAP4 antibodies).

[0089] As used herein, all references to the terms "antibody" or "antigen-binding fragment thereof" refer to the antibodies or antigen-binding fragments thereof described in this disclosure.

[0090] A "fragment" of a (conventional) antibody contains a portion of the complete antibody, particularly the antigen-binding region or variable region. Examples of antibody fragments include Fv, Fab, F(ab')2, Fab', dsFv, (dsFv)2, scFv, sc(Fv)2, double-chain antibodies, bispecific antibodies formed from antibody fragments, and multispecific antibodies. Fragments of conventional antibodies can also be single-domain antibodies, such as heavy-chain antibodies or VHH.

[0091] The term "Fab" refers to an antibody fragment with a molecular weight of approximately 50,000 and antigen-binding activity, in which about half of the N-terminal side of the heavy chain is linked to the entire light chain via disulfide bonds. It is typically obtained from fragments of IgG obtained by treating IgG with a protease (papain).

[0092] The term "F(ab')2" refers to an antibody fragment with a molecular weight of approximately 100,000 and antigen-binding activity, slightly larger than two identical Fab fragments bound by disulfide bonds in the hinge region. It is typically obtained from fragments derived by treating IgG with a protease (pepsin).

[0093] The term "Fab'" refers to an antibody fragment with a molecular weight of approximately 50,000 and antigen-binding activity, which is obtained by cleaving the disulfide bonds in the hinge region of F(ab')2.

[0094] Single-chain Fv (“scFv”) polypeptides are covalently linked VH::VL heterodimers, typically expressed by gene fusions comprising VH and VL encoding genes linked via peptide-coding linkers. The human scFv fragments of this disclosure comprise CDRs maintained in an appropriate conformation (particularly through the use of gene recombination techniques). Bivalent and multivalent antibody fragments can be spontaneously formed through the association of monovalent scFvs, or generated by coupling monovalent scFvs, such as divalent sc(Fv)2, via peptide linkers. “dsFv” refers to a VH::VL heterodimer stabilized by disulfide bonds. “(dsFv)2” represents two dsFvs coupled via peptide linkers.

[0095] The term "bispecific antibody" or "BsAb" refers to an antibody that combines the antigen-binding sites of two antibodies within a single molecule. Therefore, a BsAb can bind to two different antigens simultaneously. Genetic engineering has been increasingly used to design, modify, and produce antibodies or antibody derivatives with desired combinations of binding properties and effector functions, as described in, for example, EP 2 050764 A1.

[0096] The term "multispecific antibody" refers to an antibody that combines the antigen-binding sites of two or more antibodies within a single molecule.

[0097] The term "double-chain antibody" refers to a small antibody fragment with two antigen-binding sites. These fragments contain a heavy chain variable domain (VH) and a light chain variable domain (VL) linked together on the same polypeptide chain (VH-VL). By using a linker that is too short to allow pairing between the two domains on the same chain, these domains are forced to pair with the complementary domain of the other chain, thus creating two antigen-binding sites.

[0098] The antibodies of the compositions described in this invention can be produced by any technique known in the art, such as, but not limited to, any chemical, biotechnological, genetic, or enzymatic techniques, used alone or in combination. The antibodies of this invention may comprise polyclonal antibodies. Methods for preparing polyclonal antibodies are known to those skilled in the art (Harlow et al., Antibodies: a Laboratory Manual, Cold Spring Harbor Laboratory Press, 2nd edition (1988)).

[0099] "A combination of antibodies or antigen-binding fragments thereof" can be, in particular, a polyclonal antibody composition or a monoclonal antibody composition, and can preferably be a polyclonal antibody composition.

[0100] The terms “polyclonal antibody” or “polyclonal antibody composition” used interchangeably herein refer to a mixture of antibodies that recognize different epitopes of a given antigen, or even different epitopes of different antigens expressed by a given cell or cell population. Polyclonal antibodies encompass antibodies contained in or alternatively derived from bodily fluids, particularly serum or plasma from non-human mammalian organisms, especially pigs. The polyclonal antibody compositions of the present invention differ from compositions containing more than one monoclonal antibody. Polyclonal rabbit anti-thymocyte globulin (ATG) is an FDA and EMA approved drug commonly used in organ transplantation as an induction agent (Lacorcia et al., Transplantation, April 15, 2009; 87(7): 966-74). Newer generations of humanized polyclonal antibodies have also been tested in humans for various indications and have shown satisfactory tolerability (Poulakou et al., Front. Immunol. April 17, 2024: 15: 1330178).

[0101] As used herein, the term "monoclonal antibody" or "mAb" refers to an antibody molecule with a single amino acid sequence that targets a specific antigen, and should not be construed as requiring that the antibody be produced by any particular method. Monoclonal antibodies can be produced from a single clone of B cells or hybridomas, but they can also be recombinant, i.e., produced through protein engineering.

[0102] The term "humanized antibody" refers to antibodies that are wholly or partially non-human in origin and that have been modified to replace certain amino acids, particularly those in the framework regions of the VH and VL domains, to avoid or minimize immune responses in humans. The constant domains of humanized antibodies are, in most cases, the human CH and CL domains.

[0103] When the term "recombinant" is applied to antibodies or their antigen-binding fragments, nucleic acid sequences, expression vectors, or host cells, it means that they are the product of a combination of in vitro cloning, restriction treatments, ligation steps, and other genetic engineering procedures.

[0104] In the context of this invention, the term "antibody" specifically covers antibodies derived from pigs, and more specifically, antibodies derived from pigs that lack at least one antigenic determinant selected from (i) N-hydroxyacetylneuraminic acid (Neu5Gc) and (ii) α-1,3-galactose, and particularly lack both antigenic determinants: N-hydroxyacetylneuraminic acid (Neu5Gc) and α-1,3-galactose; and further preferably, it comprises at least one sugar moiety different from the antigenic determinants (i) N-hydroxyacetylneuraminic acid (Neu5Gc) and / or (ii) α-1,3-galactose.

[0105] The combination of antibodies used according to the invention, and more specifically the polyclonal antibody composition used according to the invention, as understood herein, particularly polyclonal antibody compositions obtainable by immunizing animals with CD3+, CD8+, TCR γ / δ-, and CD34- human tumor T cells. CD3+, CD8+, TCR γ / δ-, and CD34- are understood to be human tumor T cells expressing CD3 and CD8 but not TCR γ / δ and CD34. In particular, the animal is a pig, and especially a pig lacking at least one gene selected from the group consisting of (i) a gene encoding a functional cytidine-5'-monophosphate N-acetylneuraminic acid hydrolase (CMAH) and (ii) a gene encoding a functional α-(1,3)-galactosyltransferase, and more particularly a pig lacking both (i) a gene encoding a functional cytidine-5'-monophosphate N-acetylneuraminic acid hydrolase (CMAH) and (ii) a gene encoding a functional α-(1,3)-galactosyltransferase.

[0106] As used in this disclosure, the term "antigen" refers to a molecule or part of a molecule that can be bound by one or more antibodies. An antigen may have one or more epitopes. For example, in the context of this disclosure, an antigen is particularly selected from the group consisting of proteins FASN, CD99, GPI, SLC3A2, GART, and CKAP4.

[0107] As used herein, the term “affinity” refers to the strength of binding between an antibody and an epitope presented on an antigen. Antibody affinity is given by the dissociation constant KD, defined as [Ab] x [Ag] / [Ab-Ag], where [Ab-Ag] is the molar concentration of the antibody-antigen complex, [Ab] is the molar concentration of unbound antibody, and [Ag] is the molar concentration of unbound antigen. The affinity constant Ka is defined as 1 / Kd. Methods for determining Ab affinity can be found, for example, Harlow et al., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY., 1988; Colligan et al., Current Protocols in Immunology, Greene Publishing Assoc. and Wiley Interscience, NY, (1992, 1993), or Müller, Methods Enzymol. 1983;92:589-601, all of which are incorporated herein by reference in their entirety. A preferred and standard method known in the art for determining mAb affinity is surface plasmon resonance measurement using a Biacore instrument (Laure et al., Curr Protoc Protein Sci. 2006 Sep; Chapter 19: Unit 19.13). For multimeric antigens, such as viral capsids, both antigen-binding sites of the antibody can bind simultaneously. The Ka and KD values ​​determined by such standard methods measure the functional affinity or avidity of the interaction. For example, when the functional binding affinity (KD) (preferably measured by surface plasmon resonance) is 10... -8 mol / L (M) or less, preferably 10 -9 M to 10 -12 When M occurs, the antibody is considered to bind to the antigen.

[0108] As used herein, “IgG” refers to a polypeptide belonging to a class of antibodies essentially encoded by the well-known immunoglobulin γ gene. In humans, IgG comprises subclasses or isotypes IgG1, IgG2, IgG3, and IgG4. In mice, IgG comprises IgG1, IgG2a, IgG2b, and IgG3. In pigs, immunoglobulins comprise class or isotypes of IgM, IgD, IgG, IgE, and IgA antibodies, and IgG isotypes comprise 11 subclasses (Butler et al., Developmental and Comparative Immunology 30 (2006) 199–221; Butler et al., Developmental and Comparative Immunology 33 (2009) 321–333). Full-length IgG consists of two pairs of identical immunoglobulin chains, each pair containing one light chain and one heavy chain. Each light chain contains immunoglobulin domains VL and CL, and each heavy chain contains immunoglobulin domains VH, Cγ1 (also known as CH1), Cγ2 (also known as CH2), and Cγ3 (also known as CH3).

[0109] As used herein, the term "antigenic determinant" (or epitope) applied to porcine antibodies refers to a structural component of an antigen molecule, including the antigenic protein and antigenic carbohydrate, responsible for its specific interaction with antibody molecules induced by the same or related antigens. By extension, the term "antigenic determinant" applied to porcine antibodies herein also refers collectively to antigen molecules containing multiple epitopes, including conformational motifs where the glycosyl portion is essential but represents only a portion of the epitope, readily recognized by antibody molecules induced by the same or related antigens. For example, the antigen molecule N-hydroxyacetylneuraminic acid (Neu5Gc) may be referred to herein as an "antigenic determinant," although the antigen molecule may exhibit more than one epitope recognized by antibodies induced by Neu5Gc or molecules containing Neu5Gc.

[0110] As used herein, “conventional polyclonal antibody” refers to a polyclonal antibody that does not lack the antigenic determinants N-hydroxyacetylneuraminic acid (Neu5Gc) and α-1,3-galactose, and particularly porcine or rabbit polyclonal antibodies. In this regard, products commercially available under the names Thymoglobulin, Grafalon, Atgam, or p-ALG® deserve special mention.

[0111] The term “cancer” is used in this article in its traditional sense, and refers to cells or groups of cells that exhibit uncontrolled growth and invasion of adjacent tissues.

[0112] This article further provides more specific examples of cancers for consideration. In this article, cancer may specifically refer to cancers selected from the group consisting of: myeloma; melanoma; skin cancer; breast cancer; brain tumors, bladder cancer, cervical cancer, prostate cancer; primary and metastatic colorectal cancer, particularly colon cancer; mesothelioma; lung cancer, particularly non-small cell lung cancer; liver cancer, particularly hepatocellular carcinoma or bile duct cancer; primary and metastatic pancreatic cancer; kidney cancer; soft tissue sarcoma; thyroid cancer; lymphoma, including Hodgkin's lymphoma and non-Hodgkin's lymphoma, particularly B-cell lymphoma or T-cell lymphoma, more particularly T-cell lymphoma; stomach cancer; head and neck cancer; ovarian cancer; sarcoma; acute or chronic leukemia, particularly T-cell leukemia or myeloid leukemia; osteosarcoma; anal cancer; testicular cancer; uterine cancer; thyroid cancer; central nervous system cancers; gastrointestinal stromal cancer; epidermal cancer; and esophageal cancer. In one specific embodiment, the cancer described in this invention is a cancer that expresses at least one antigen selected from the group consisting of FASN, CD99, GPI, SLC3A2, GART and CKAP4, particularly the group consisting of GPI and GART.

[0113] In a specific implementation plan, the cancer may be selected from the group consisting of: myeloma; melanoma; skin cancer; breast cancer; brain tumors, bladder cancer, cervical cancer, prostate cancer; primary and metastatic colorectal cancer, especially colon cancer; mesothelioma; lung cancer, especially non-small cell lung cancer; liver cancer, especially hepatocellular carcinoma or bile duct cancer; primary and metastatic pancreatic cancer; kidney cancer; soft tissue sarcoma; thyroid cancer; lymphoma, including Hodgkin lymphoma and non-Hodgkin lymphoma, especially B-cell lymphoma. Tumors or T-cell lymphomas, especially T-cell lymphomas; gastric cancer; head and neck cancer; ovarian cancer; sarcoma; acute or chronic leukemia, especially T-cell or myeloid leukemia; osteosarcoma; anal cancer; testicular cancer; uterine cancer; thyroid cancer; central nervous system cancers; gastrointestinal stromal cancers; epidermal cancers; and esophageal cancers, which express at least one antigen selected from the group consisting of FASN, CD99, GPI, SLC3A2, GART, and CKAP4, especially the group consisting of GPI and GART.

[0114] The term "therapeuticly effective amount" for the combination of antibodies used according to the present invention means an amount of antibody sufficient to achieve the desired effect described herein, particularly for treating human subjects with cancer. However, it should be understood that the total daily dosage of the combination of antibodies implemented according to the present invention will be determined by the attending physician within reasonable medical judgment. The specific therapeutically effective dose level for any particular patient will depend on a variety of factors, including: the nature of the cancer being treated; the activity of the specific antibody used; the specific composition used; the patient's age, weight, general health condition, sex, and diet; the timing, route of administration, and excretion rate of the specific antibody used; the duration of treatment; drugs used in combination with or concurrently with the specific antibody used; and similar factors known in the medical field. For example, it is known to those skilled in the art that administration of the compound is initiated at a level below that required to achieve the desired therapeutic effect, and the dose is gradually increased until the desired effect is achieved.

[0115] The terms "treatment," "treatment," or "therapy" refer to the application of an active agent to achieve the following purposes: cure, heal, alleviate, relieve, alter, remedy, improve, enhance, or influence a condition (i.e., cancer), cancer symptoms, or prevent or delay the occurrence of symptoms, complications, biochemical indicators of the disease, or otherwise prevent or inhibit the further development of cancer. In particular, in the context of this invention, treating cancer includes providing antitumor activity. Specifically, antitumor activity comprises complement-dependent cytotoxicity (CDC), complement-dependent cytotoxicity (ADCC) from killer cells, apoptotic activity, and / or complement-dependent cytotoxicity (ADCP) from phagocytes, and more particularly, antitumor activity consisting of complement-dependent cytotoxicity (CDC), complement-dependent cytotoxicity (ADCC) from killer cells, apoptotic activity, and complement-dependent cytotoxicity (ADCP) from phagocytes.

[0116] The terms “wild-type animal” or “WT animal” are used in this document in contrast to genetically modified animals. For example, “wild-type pig” refers to a pig that does not lack at least one gene selected from the group consisting of (i) a gene encoding a functional cytidine-5'-monophosphate N-acetylneuraminic acid hydrolase (CMAH) and (ii) a gene encoding a functional α-(1,3)-galactosyltransferase.

[0117] As a suitable method for obtaining polyclonal antibodies that may be present in the antibody combination according to the present invention, it is particularly worth mentioning the method of fractionation precipitation with ethanol, ammonium sulfate, rivanol, polyethylene glycol or caprylic acid, followed by flow through an ion exchange column; other methods may involve protein A affinity columns or protein G affinity columns. The obtained antibodies can then be routinely processed for their intravenous administration, for example by enzymatic digestion with plasmin, papain or pepsin. In this regard, the scheme implemented in Example 3 of EP 0 335 804, which implements DEAE cellulose ion exchange chromatography, can be referred to more specifically.

[0118] Such antibodies can be generated, for example, by immunizing non-human animals (particularly non-human mammals) according to methods known to those skilled in the art. Non-human mammals can be selected from the group consisting of: rodents, such as mice, rats, guinea pigs, and hamsters; lagomorphs, such as rabbits; ferrets; felines, such as cats; canines, such as dogs; goats; sheep; bovids, such as dairy cows; suidae, such as pigs and fattening pigs; camels; horses; and non-human primates. Non-human mammals can be more particularly pigs. Thus, for example, "pig-derived anticancer pAb" means a polyclonal antibody obtained by immunizing pigs with a combination of proteins of interest or by using cells that naturally express or are genetically modified to express proteins of interest on their cell surface, the proteins of interest being the targets of the polyclonal antibody to be obtained. See Reynard et al., pLoS One. 2016; 11(6): e0156775; Schieferdecker et al., Oncotarget. Oct 11, 2016; 7(41): 67061–67070 and Zhang et al. 2014 (DOI: 10.1038 / srep04984).

[0119] The terms “peptide” and “protein” are used interchangeably in this document and refer to a sequence of amino acid residues. These terms apply to amino acid sequences in which one or more amino acid residues are artificial chemical mimics of naturally occurring amino acids, as well as to both naturally occurring and non-natural amino acid sequences.

[0120] 2. The antibody combination according to the present invention.

[0121] As described above, the present invention relates to the use of a combination of antibodies or antigen-binding fragments thereof for the treatment of cancer in human subjects in need.

[0122] Such combinations include:

[0123] - At least a first antibody or its antigen-binding fragment thereof, wherein the first antibody or its antigen-binding fragment specifically binds to the FASN (FasN fragment);

[0124] - At least a second antibody or its antigen-binding fragment, wherein the second antibody or its antigen-binding fragment specifically binds to CD99 (differentiation cluster 99); and

[0125] - At least a third antibody or its antigen-binding fragment thereof, said third antibody or its antigen-binding fragment specifically binding to an antigen selected from the group consisting of GPI (glucose-6-phosphate isomerase), SLC3A2 (solute carrier family 3 member 2), GART (glycamide ribonucleotide formyltransferase) and CKAP4 (cytoskeleton-associated protein 4), particularly the group consisting of GPI and GART.

[0126] Each antibody present in the combination used according to the present invention can be independently a monoclonal antibody, a polyclonal antibody, or an antigen-binding fragment thereof. As defined above, the antigen-binding fragment can be independently selected from the group consisting of Fv, Fab, F(ab')2, Fab', dsFv, (dsFv)2, scFv, sc(Fv)2, or double-chain antibodies.

[0127] The antibody combination used according to the present invention may comprise antibodies targeting more than three different proteins from the group consisting of FASN, CD99, GPI, SLC3A2, GART, and CKAP4. Therefore, the antibodies present in the antibody combination used according to the present invention may target at least three proteins, four proteins, at least four proteins, five proteins, at least five proteins, six proteins, at least six proteins, seven proteins, at least seven proteins, eight proteins, at least eight proteins, nine proteins, or at least nine proteins from the group consisting of FASN, CD99, GPI, SLC3A2, GART, and CKAP4.

[0128] In particular, the combination of antibodies used according to the present invention comprises antibodies targeting three, four, or five, more particularly four or five proteins selected from the group consisting of FASN, CD99, GPI, SLC3A2, GART, and CKAP4.

[0129] Some or all of the antibodies used in the antibody combination according to the invention may lack at least one antigenic determinant selected from (i) N-hydroxyacetylneuraminic acid (Neu5Gc) and (ii) α-1,3-galactose. Some or all of the antibodies used in the invention may in particular lack two antigenic determinants: N-hydroxyacetylneuraminic acid (Neu5Gc) and α-1,3-galactose.

[0130] Methods that allow for the identification or characterization of such antibodies are common knowledge to those skilled in the art. Methods that those skilled in the art can use to identify or characterize the antibodies described in this invention include enzyme-linked immunosorbent assay (ELISA), wherein, for example, anti-Neu5Gc antibodies and anti-Gal antibodies are used as detection molecules.

[0131] The antibody from the antibody combination according to the present invention may be an immunoglobulin G antibody.

[0132] For example, the structure and genetics of porcine IgG have been described in the literature (Butler et al., 2009 - DOI:10.1007 / s00251-009-0356-0). Based on the analysis of the IgH locus sequence in the genome, 11 isotypes have been proposed, named IgG1a, IgG1b, IgG2a, IgG2b, IgG3, IgG4a, IgG4b, IgG5a, IgG5b, IgG6a, and IgG6b. Based on existing knowledge regarding the relative abundance of porcine IgG subclasses and the affinity of the Fc domain for protein A (Butler et al., 2009 - DOI:10.1007 / s00251-009-0356-0), the relative composition of IgG isotypes in porcine DKO polyclonal antibodies is estimated to be >80% porcine IgG1a / b, 11% IgG2a / b, 5.5% IgG3, 3% IgG4a / b, with the remainder being other isotypes (IgG5-6). After purification using protein A chromatography, no detectable IgM or IgA isotypes were found in the porcine DKO IgG polyclonal antibody formulation.

[0133] Fatty acid synthase (FASN) is a multi-enzyme protein that acts as a key regulator of lipid metabolism, especially fatty acid synthesis.

[0134] The amino acid sequence of FASN is referenced from UNIPROT A0A0U1RQF0.

[0135] Antibodies that specifically target FASN (anti-FASN antibodies) are well known in the art. For example, monoclonal antibody 15973814 (commercialized by Fischer Scientific), polyclonal antibody LS-B3636 (commercialized by LifeSpan BioSciences), or polyclonal antibody GTX109833 (commercialized by GeneTex) may be mentioned.

[0136] The protein CD99 (differentiation cluster 99) is a glycosylated transmembrane protein involved in many important cellular functions, such as cell adhesion and migration, cell death and differentiation, intracellular protein transport, endocytosis and exocytosis.

[0137] The amino acid sequence of CD99 is referenced in UniProt P14209.

[0138] Antibodies that specifically target CD99 (anti-CD99 antibodies) are well known in the art. For example, monoclonal antibody AA 23-122 may be mentioned. For further information on such antibodies, see application WO2015161267.

[0139] Several studies have demonstrated the potential of targeting CD99 in T-cell acute lymphoblastic leukemia (ALL) with varying degrees of success, attributable to the high overexpression of CD99 in T-ALL compared to normal T cells (Ebadi et al., Scientific Reports 2021, 11:24374; Kotemul et al., Exploration of Targeted Anti-tumor Therapy 2024, 5:96-107). However, targeting CD99 with monoclonal antibodies appears to be quite challenging, especially due to the presence of different isotypes. Co-expression of CD99LF and CD99SF induces apoptosis, while CD99LF alone induces cell aggregation without inducing apoptosis (Alberti et al., FASEB 2002, Dec;16(14):1946-8). Furthermore, numerous epitopes crucial for inducing apoptosis have been identified, with apoptosis occurring via either Fas-dependent or Fas-independent pathways, highlighting the complex nature of CD99-mediated cell death mechanisms (Pettersen et al., J Immunol 2001, 166(8): 4931-4942). Moreover, effective apoptosis induction appears to depend on CD99 aggregation, requiring antibody titers of 3 or higher to achieve optimal therapeutic efficacy (Romero et al., J of Molecular Biology 2022, 167402).

[0140] GPIs (glucose-6-phosphate isomerases) are identified as moonlighting proteins based on their ability to perform different functions, such as interconverting glucose-6-phosphate (G6P) and fructose-6-phosphate (F6P) as glycolytic enzymes (glucose-6-phosphate isomerases), promoting the survival of skeletal muscle motor neurons and sensory neurons as neurotrophic factors, and inducing immunoglobulin secretion as lymphokines.

[0141] The amino acid sequence of GPI is referenced from UniProt P06744.

[0142] Antibodies that specifically target GPI (anti-GPI antibodies) are well known in the art. The monoclonal antibody MABN691 (commercialized by Merck Millipore) may be mentioned. For further information on such antibodies, see application WO2000064469.

[0143] The protein SLC3A2 belongs to the SLC (solute carrier) protein family, which consists of a superfamily of 65 members. They are primarily involved in molecular / drug delivery and are frequently cited research resources in drug resistance studies. The SLC family is subclassified according to the type of molecular delivery they are responsible for, including the SLC1 family, SLC2 family, and so on, up to the SLC65 family. Furthermore, each family has sub-members. For example, the SLC3 family has two members: SLC3a1 and SLC3a2. Typically, in their mechanism of action, they act together with members of the SLC7 family, so most studies usually analyze both together. On the other hand, SLC3a2 forms a dimer with SLC7a5 to achieve its function and together they participate in amino acid delivery. CD98 is a transmembrane protein located on the cell surface. Therefore, it can be easily targeted by drugs.

[0144] The amino acid sequence of SLC3A2 is referenced in GenBank: KAI2560544.1.

[0145] Antibodies that specifically target SLC3A2 (anti-SLC3A2 antibodies) are well known in the art. For example, mention may be made of the monoclonal antibody sc-390154 (commercialized by Santa Cruz Biotechnology), MA5-29573 (commercialized by ThermoFischer Scientific), or the polyclonal antibody 15193-1-AP (commercialized by Proteintech Group Inc.). Application WO20172114458 also describes several antibodies targeting SLC3A2 and is incorporated herein by reference.

[0146] The protein GART (glycamide ribonucleotide formyltransferase) catalyzes the N-formylation of glycamide ribonucleotides. GART is an important step in purine nucleotide synthesis and a target for blocking the proliferation of malignant cells.

[0147] The amino acid sequence of GART is referenced from UniProt Q71VH3.

[0148] Anti-GART antibodies that specifically target GART are well known in the field. See Joe Dotzlaf et al. (Hybridoma (Larchmt). 2006; 25:139-44).

[0149] The protein CKAP4 (cytoskeleton-associated protein 4) is the receptor for Dickkopf1 (DKK1), a secreted protein that antagonizes oncogenic Wnt signaling by binding to the Wnt co-receptor low-density lipoprotein receptor-associated protein 6 (LRP6) and can also regulate its own signaling to promote cancer cell proliferation.

[0150] The amino acid sequence of CKAP4 is referenced from the NCBI reference sequence: NP_006816.2.

[0151] Antibodies that specifically target CKAP4 (anti-CKAP4 antibodies) are well known in the art. For example, the polyclonal antibody 16686-1-AP (commercialized by Proteintech Group Inc.), the monoclonal antibody MOB-3287z (commercialized by Creative Biolabs), or the monoclonal antibody sc-393544 (commercialized by Santa Cruz Biotechnology) may be mentioned. Several antibodies targeting CKAP4 are also described in application WO2019065747, which is incorporated herein by reference.

[0152] The combination of antibodies used according to the present invention may comprise at least a fourth antibody or its antigen-binding fragment, said fourth antibody or its antigen-binding fragment specifically binding to an antigen selected from the group consisting of: GPI, SLC3A2, GART, and CKAP4. The antigen targeted by said at least the fourth antibody may be particularly different from the three antigens bound by the first, second, and third antibodies or their antigen-binding fragments.

[0153] Therefore, the combination of antibodies or antigen-binding fragments thereof used according to the present invention may include:

[0154] - At least a first antibody or its antigen-binding fragment thereof, wherein the first antibody or its antigen-binding fragment specifically binds to the FASN (FasN fragment);

[0155] - At least a second antibody or its antigen-binding fragment, wherein the second antibody or its antigen-binding fragment specifically binds to CD99 (differentiation cluster 99); and

[0156] - At least a third antibody or its antigen-binding fragment, said third antibody or its antigen-binding fragment specifically binding to an antigen selected from GPI (glucose-6-phosphate isomerase), SLC3A2 (solute carrier family 3 member 2), GART (glycamide ribonucleotide formyltransferase), and CKAP4 (cytoskeleton-associated protein 4), particularly the group consisting of GPI and GART, and

[0157] - At least a fourth antibody or its antigen-binding fragment thereof, said fourth antibody or its antigen-binding fragment thereof specifically binding to an antigen selected from the group consisting of GPI (glucose-6-phosphate isomerase), SLC3A2 (solute carrier family 3 member 2), GART (glycamide ribonucleotide formyltransferase) and CKAP4 (cytoskeleton-associated protein 4), particularly the group consisting of GPI and GART, said fourth antibody or its antigen-binding fragment thereof being different from the third antibody or its antigen-binding fragment thereof.

[0158] In one specific embodiment, the combination of antibodies used according to the present invention comprises:

[0159] -At least a first antibody or its antigen-binding fragment thereof, wherein the first antibody or its antigen-binding fragment specifically binds to FASN;

[0160] -At least a second antibody or its antigen-binding fragment thereof, wherein the second antibody or its antigen-binding fragment specifically binds to CD99;

[0161] - At least a third antibody or its antigen-binding fragment, said third antibody or its antigen-binding fragment specifically binding to GPI; and

[0162] - At least a fourth antibody or its antigen-binding fragment, said fourth antibody or its antigen-binding fragment specifically binds to GART.

[0163] Examples of such combinations are shown in the embodiments of this specification under the name PA1.

[0164] As previously stated, in all these embodiments, the antibodies in the combination, particularly the polyclonal antibodies in the combination, may independently lack at least one antigenic determinant selected from (i) N-hydroxyacetylneuraminic acid (Neu5Gc) and (ii) α-1,3-galactose, and may more particularly lack two antigenic determinants: N-hydroxyacetylneuraminic acid (Neu5Gc) and α-1,3-galactose.

[0165] In one specific embodiment, in all the above embodiments, the antibody in the combination, particularly the polyclonal antibody, lacks at least one antigenic determinant selected from (i) N-hydroxyacetylneuraminic acid (Neu5Gc) and (ii) α-1,3-galactose, and more particularly lacks both antigenic determinants: N-hydroxyacetylneuraminic acid (Neu5Gc) and α-1,3-galactose.

[0166] The antibodies used in the combination according to the invention, and particularly the polyclonal antibodies used according to the invention, can be of any origin, such as those derived from non-human mammals or synthetic, monoclonal or polyclonal. Such non-human mammals can be selected from the group consisting of: rodents, such as mice, rats, guinea pigs, and hamsters; lagomorphs, such as rabbits; ferrets; felines, such as cats; canines, such as dogs; goats; sheep; bovids, such as cows; suidae, such as pigs and fattening pigs; camels; horses; and non-human primates.

[0167] The antibodies used in the combination according to the invention, and especially the polyclonal antibodies used according to the invention, are preferably derived from pigs, and particularly from pigs lacking at least one gene selected from the group comprising (i) a gene encoding a functional cytidine-5'-monophosphate N-acetylneuraminic acid hydrolase (CMAH) and (ii) a gene encoding a functional α-(1,3)-galactosyltransferase, and more particularly from pigs lacking both (i) a gene encoding a functional cytidine-5'-monophosphate N-acetylneuraminic acid hydrolase (CMAH) and (ii) a gene encoding a functional α-(1,3)-galactosyltransferase.

[0168] In particular, the combination used according to the present invention can be a combination of monoclonal antibodies or a polyclonal antibody composition, and preferably a polyclonal antibody composition.

[0169] In one specific embodiment, the combination used according to the invention comprises a pharmaceutical composition further comprising a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are further defined above.

[0170] The pharmaceutical composition may be in liquid form.

[0171] The pharmaceutical composition may be in solid form, including lyophilized form.

[0172] Pharmaceutical compositions can be formulated according to standard methods, such as those described in Remington: The Science and Practice of Pharmacy (Lippincott Williams & Wilkins; 21st edition, 2005).

[0173] The pharmaceutical composition may further comprise at least one additional anticancer drug that is different from the antibody in the combination of antibodies.

[0174] The term "combination of antibodies" is also used in this article to refer to "a combination of antibodies or antigen-binding fragments thereof".

[0175] For example, other anticancer drugs can be selected from groups composed of free monoclonal antibodies, especially groups composed of the following monoclonal antibodies: anti-CD19, anti-CD20, anti-CD30, anti-CD137, anti-CTLA4, anti-TIM-3, anti-B7-H3, anti-CD123, anti-CD134, anti-CD154, anti-LAG-3, anti-CD227, anti-BTNA3, anti-CD39, anti-CD73, anti-CD115, anti-CD47, anti-SIRPα, anti-SIRPγ, anti-CD28, anti-NCR, anti-NKp46, anti-NKp30, anti-NKp44, anti-NKG2D, anti-PD1, anti-PDL1, moglizazumab, oxotuzumab, velpotuzumab, yttrium Y 90-tiemotuzumab, motuzumab, and anti-DNAM-1 monoclonal antibody.

[0176] The pharmaceutical composition may further include at least one chemotherapy treatment.

[0177] Chemotherapy treatment can be selected from those known in the art. In particular, those skilled in the art will know how to tailor the choice of chemotherapy according to the cancer to be treated. In one specific embodiment, chemotherapy treatment is selected from the group consisting of: chemotherapy regimens consisting of cyclophosphamide, daunorubicin, vincristine, and prednisone (CHOP); chemotherapy regimens consisting of cyclophosphamide, daunorubicin, vincristine, etoposide, and prednisone (CHOEP); HDAC inhibitors; ibrutinib, acalabrutinib, zanubrutinib, curapannisi, and veneclade.

[0178] As defined herein, the dosage range of the combination used according to the invention can be from 0.001 to 100 mg or more per kg body weight (mg / kg) or higher, for example 0.1, 1.0, 10 or 50 mg / kg body weight, preferably 1 to 20 mg / kg. The dosage and frequency of administration can be adjusted as detailed previously.

[0179] Furthermore, any routine procedures can be performed after any injection of the combination used according to the present invention to prevent and / or avoid allergic reactions.

[0180] Furthermore, the injection of the combination or composition according to the present invention can be performed through a large peripheral passage, or, if possible, through a central catheter.

[0181] As is known in the art, adjustments may be required for protein degradation, systemic versus local delivery, as well as for age, weight, general health status, sex, diet, timing of administration, possible allergies, drug interactions, and severity of the condition, and these adjustments can be readily determined by routine experiments performed by those skilled in the art.

[0182] The combination or composition used according to the present invention can be administered in a variety of ways, including but not limited to oral, subcutaneous, intravenous, parenteral, intranasal, intra-inhalation (such as nebulization or intratracheal spray), intra-aortic, intraocular, intradermal, vaginal, transdermal, topical (e.g., gel), intraperitoneal, intramuscular, intrapulmonary, or intrathecal administration.

[0183] The application of the combination or composition of the present invention can be carried out according to the Besredka method.

[0184] The combinations or compositions of the present invention may be in forms particularly suitable for intravenous administration.

[0185] 3. Implementation of the antibody or combination or composition thereof described in this invention.

[0186] The antibody or combination thereof, or a composition comprising the antibody, as described in this invention, is used in a therapeutically effective amount.

[0187] As previously stated, the use of combinations of antibodies or their compositions for the treatment of cancer in human subjects.

[0188] Cancer can be specifically selected from the following groups: myeloma; melanoma; skin cancer; breast cancer; brain tumors, bladder cancer, cervical cancer, prostate cancer; primary and metastatic colorectal cancer, especially colon cancer; mesothelioma; lung cancer, especially non-small cell lung cancer; liver cancer, especially hepatocellular carcinoma or bile duct cancer; primary and metastatic pancreatic cancer; kidney cancer; soft tissue sarcoma; thyroid cancer; lymphoma, including Hodgkin's lymphoma and non-Hodgkin's lymphoma, especially B-cell lymphoma or T-cell lymphoma, and even more so T-cell lymphoma; stomach cancer; head and neck cancer; ovarian cancer; sarcoma; acute or chronic leukemia, especially T-cell leukemia or myeloid leukemia; osteosarcoma; anal cancer; testicular cancer; uterine cancer; thyroid cancer; central nervous system cancers; gastrointestinal stromal cancer; epidermal cancer; and esophageal cancer.

[0189] In one specific embodiment, the cancer described in this invention is a cancer that expresses at least one antigen selected from the group consisting of FASN, CD99, GPI, SLC3A2, GART and CKAP4, particularly the group consisting of GPI and GART.

[0190] In a specific implementation plan, cancer can be selected from the group consisting of: myeloma; melanoma; skin cancer; breast cancer; brain tumors, bladder cancer, cervical cancer, prostate cancer; primary and metastatic colorectal cancer, especially colon cancer; mesothelioma; lung cancer, especially non-small cell lung cancer; liver cancer, especially hepatocellular carcinoma or bile duct cancer; primary and metastatic pancreatic cancer; kidney cancer; soft tissue sarcoma; thyroid cancer; lymphoma, including Hodgkin lymphoma and non-Hodgkin lymphoma, especially B-cell lymphoma. Tumors or T-cell lymphomas, especially T-cell lymphomas; gastric cancer; head and neck cancer; ovarian cancer; sarcoma; acute or chronic leukemia, especially T-cell or myeloid leukemia; osteosarcoma; anal cancer; testicular cancer; uterine cancer; thyroid cancer; central nervous system cancers; gastrointestinal stromal cancers; epidermal cancers; and esophageal cancers, which express at least one antigen selected from the group consisting of FASN, CD99, GPI, SLC3A2, GART, and CKAP4, especially the group consisting of GPI and GART.

[0191] In one specific implementation, the combination used according to the present invention is as follows:

[0192] -At least a first antibody or its antigen-binding fragment thereof, wherein the first antibody or its antigen-binding fragment specifically binds to FASN;

[0193] -At least a second antibody or its antigen-binding fragment, wherein the second antibody or its antigen-binding fragment specifically binds to CD99; and

[0194] - At least a third antibody or its antigen-binding fragment, said third antibody or its antigen-binding fragment specifically binds to an antigen selected from the group consisting of GPI, SLC3A2, GART and CKAP4, particularly the group consisting of GPI and GART;

[0195] Cancers are selected from the following groups: myeloma; melanoma; skin cancer; breast cancer; brain tumors, bladder cancer, cervical cancer, prostate cancer; primary and metastatic colorectal cancer, especially colon cancer; mesothelioma; lung cancer, especially non-small cell lung cancer; liver cancer, especially hepatocellular carcinoma or bile duct cancer; primary and metastatic pancreatic cancer; kidney cancer; soft tissue sarcoma; thyroid cancer; lymphoma, including Hodgkin's lymphoma and non-Hodgkin's lymphoma, especially B-cell lymphoma or T-cell lymphoma, and more particularly T-cell lymphoma; stomach cancer; head and neck cancer; ovarian cancer; sarcoma; acute or chronic leukemia, especially T-cell leukemia or myeloid leukemia; osteosarcoma; anal cancer; testicular cancer; uterine cancer; thyroid cancer; central nervous system cancers; gastrointestinal stromal cancer; epidermal cancer; and esophageal cancer.

[0196] In one specific implementation plan, the cancer is lymphoma.

[0197] In particular, cancer is:

[0198] - Select lymphomas from the following groups:

[0199] (i) Non-Hodgkin lymphoma selected from the group consisting of the following:

[0200] - B-cell lymphomas, particularly those selected from the group consisting of: diffuse large B-cell lymphoma, follicular lymphoma, small lymphocytic lymphoma (or chronic lymphocytic leukemia), mantle cell lymphoma, marginal zone B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma (or Waldenström macroglobulinemia), hairy cell leukemia, and primary central nervous system lymphoma; and

[0201] -T-cell lymphomas, particularly those selected from the group consisting of: precursor T-lymphoblastic lymphoma (or precursor T-lymphoblastic leukemia), peripheral T-cell lymphoma, and cutaneous T-cell lymphoma; and

[0202] (ii) Hodgkin lymphoma selected from the following groups: tuberous sclerosis Hodgkin lymphoma, mixed cellularity Hodgkin lymphoma, lymphocytic depletion Hodgkin lymphoma, and lymphocyte-rich Hodgkin lymphoma; or

[0203] - Leukemia, particularly those selected from the following groups: acute myeloid leukemia (AML), chronic myeloid leukemia (CML), and histiocytic leukemia.

[0204] In one specific implementation plan, cancer is selected from the group consisting of the following:

[0205] -T-cell acute lymphoblastic leukemia (T-ALL);

[0206] - Peripheral T-cell lymphoma (PTCL), especially PTCL selected from the following groups: PTCL nonspecific type (PTCL-NOS), enteropathy-associated T-cell lymphoma (EATL), monomorphic epithelial-friendly intestinal T-cell lymphoma (MEITL), anaplastic large cell lymphoma (ALCL) and angioimmunoblastic T-cell lymphoma (AITL), extranodal NK / T-cell lymphoma nasal type (ENKL), hepatosplenic γδ T-cell lymphoma (HSGDTCL), intestinal T-cell lymphoma (ITCL), mycosis fungoides and Cezari syndrome;

[0207] - Cutaneous T-cell lymphoma (CTCL); and

[0208] -T-cell lymphoblastic lymphoma (T-LBL).

[0209] This article further provides the use of antibodies or combinations of antigen-binding fragments thereof prepared by immunizing animals with CD3+, CD8+, TCR γ / δ- and CD34- human tumor T cells in the preparation of pharmaceuticals.

[0210] This article further provides a method for treating cancer in an individual in need, the method comprising at least the step of administering to said individual a combination of an antibody or an antigen-binding fragment thereof, the combination comprising:

[0211] -At least a first antibody or its antigen-binding fragment thereof, wherein the first antibody or its antigen-binding fragment specifically binds to FASN;

[0212] -At least a second antibody or its antigen-binding fragment, wherein the second antibody or its antigen-binding fragment specifically binds to CD99; and

[0213] - At least a third antibody or its antigen-binding fragment, said third antibody or its antigen-binding fragment specifically binding to an antigen selected from the group consisting of GPI, SLC3A2, GART and CKAP4, particularly the group consisting of GPI and GART.

[0214] Individuals in need are those with cancer, particularly the cancers described above. In one specific implementation, individuals in need are those with lymphoma, particularly those with PTCL.

[0215] This article further describes a method for treating cancer in individuals in need, which includes:

[0216] Animals were immunized with CD3+, CD8+, TCR γ / δ- and CD34- human tumor T cells;

[0217] Isolation of antibodies or combinations thereof from immunized animals; and

[0218] A composition comprising a combination of isolated antibodies or antigen-binding fragments thereof, administered to an individual in a therapeutically effective amount.

[0219] The combination of antibodies or their antigen-binding fragments includes:

[0220] -At least a first antibody or its antigen-binding fragment thereof, wherein the first antibody or its antigen-binding fragment specifically binds to FASN;

[0221] -At least a second antibody or its antigen-binding fragment, wherein the second antibody or its antigen-binding fragment specifically binds to CD99; and

[0222] - At least a third antibody or its antigen-binding fragment, said third antibody or its antigen-binding fragment specifically binding to an antigen selected from the group consisting of GPI, SLC3A2, GART and CKAP4, particularly the group consisting of GPI and GART.

[0223] This article further describes a method for preparing an antibody or a combination of antigen-binding fragments thereof, the method comprising at least the following steps:

[0224] Animals were immunized with CD3+, CD8+, TCR γ / δ-, and CD34- human tumor T cells; and

[0225] A combination of antibodies or antigen-binding fragments isolated from immunized animals.

[0226] This article also describes: a pharmaceutical composition for treating cancer, the pharmaceutical composition comprising a combination of antibodies or antigen-binding fragments thereof, said combination of antibodies or antigen-binding fragments thereof comprising:

[0227] -At least a first antibody or its antigen-binding fragment thereof, wherein the first antibody or its antigen-binding fragment specifically binds to FASN;

[0228] -At least a second antibody or its antigen-binding fragment, wherein the second antibody or its antigen-binding fragment specifically binds to CD99; and

[0229] - At least a third antibody or its antigen-binding fragment, said third antibody or its antigen-binding fragment specifically binding to an antigen selected from the group consisting of GPI, SLC3A2, GART and CKAP4, particularly the group consisting of GPI and GART.

[0230] Combinations of antibodies or their antigen-binding fragments and cancer can be defined as above.

[0231] This disclosure is further illustrated by the following embodiments, but is not limited in any way to these embodiments.

[0232] Example

[0233] Example 1: In vitro cytotoxicity study (CDC assay) of the antibody combination of the present invention in different human cell lines.

[0234] Different blood cancer cell lines were exposed to a combination of antibodies of the present invention (i.e., a polyclonal antibody composition named “PA1” at a concentration of 100 μg / mL) in the presence of rabbit complement (final dilution 1 / 3) (n=3).

[0235] The liquid cancer cell lines tested are as follows:

[0236] -HPB-ALL, Jurkat, and T1301 cell lines represent T-ALL;

[0237] - The HUT-78 cell line represents cutaneous T-cell lymphoma (Cezari syndrome); and

[0238] The -OCI-LY-12 cell line represents PTCL-NOS and has been described as resistant to CHOP (Magni et al., 2019).

[0239] After incubation at 37°C for 30 minutes, use NucleoCounter® NC-3000 TM Advanced imaging cell analyzer (Chemometec, Denmark) measures cell viability.

[0240] Peripheral blood mononuclear cells (PBMCs) isolated from three different healthy donors were exposed to the same concentration of PA1 at 37°C for 30 minutes in the presence of rabbit complement (final dilution 1 / 3). NucleoCounter® NC-3000 was used. TM Cell viability was measured using an advanced imaging cell analyzer (Chemometec, Denmark).

[0241] result:

[0242] The results showed that PA1 induced strong cytotoxicity of 62% to 92% against T-ALL cell lines, and approximately 40% against cutaneous T-cell lymphoma cell lines and PTCL NOS cell lines (see [link to study]). Figure 1 ).

[0243] Specifically, the results showed that PA1 specifically targeted and killed tumor cells without damaging healthy PBMCs. No cross-reactivity with healthy PBMCs was observed.

[0244] Example 2: In vitro antitumor effect of the antibody combination of the present invention in the CHOP-resistant PTCL-NOS human cell line. Tumor activity (CDC assay)

[0245] Serial dilutions of the polyclonal antibody composition PA1 (from 10 µg / ml to 300 µg / ml) were incubated with OCI-LY-12 cells, representing PTCL-NOS cancer, at 37°C for 24 h in the presence of rabbit complement (final dilution 1 / 3). Incubation with rabbit complement for 24 h was used to assess the overall antitumor activity of the PA1 polyclonal antibody composition (complement-dependent cytotoxicity (CDC) and apoptosis). Note that PTCL-NOS cell lines are resistant to standard CHOP therapy approved for the treatment of PTCL-NOS cancer (Sibon et al., 2022; Deng et al., 2019; Ruan et al., 2023).

[0246] After incubation at 37°C for 24 hours, NucleoCounter® NC-3000 was used.TM Advanced imaging cell analyzer (Chemometec, Denmark) measures cell viability.

[0247] result:

[0248] The results showed that PA1 induced strong antitumor activity in the OCI-LY-12 cell line, with a cell death rate of 80% at 30 µg / ml (see [link to study]). Figure 2 ).

[0249] Example 3: Comparison of the cytotoxic activity of the antibody combination of the present invention and alemtuzumab in different human cell lines. (Measured by CDC)

[0250] The complement-dependent cytotoxicity (CDC) of the polyclonal antibody composition PA1 of the present invention in liquid cancer cell lines was compared with that of the anti-CD52 monoclonal antibody alemtuzumab.

[0251] Cytotoxic activity was evaluated on three T-ALL cell lines (HPB-ALL, Jurkat, and T1301) and one cutaneous T-lymphoma cell line (HUT-78). Polyclonal antibody composition (PA1) and monoclonal antibody (alemumab) were incubated with these different cell lines at 37°C for 30 min at the same concentration of 100 µg / ml in the presence of rabbit complement (final dilution to 1 / 3) (100,000 tumor cells per well).

[0252] After incubation at 37°C for 30 minutes, use NucleoCounter® NC-3000 TM Advanced imaging cell analyzer (Chemometec, Denmark) measures cell viability.

[0253] Peripheral blood mononuclear cells (PBMCs) isolated from three different healthy donors were exposed to the same concentrations of PA1 and alemtuzumab for 30 minutes at 37°C in the presence of rabbit complement (final dilution 1 / 3). NucleoCounter® NC-3000 was also used. TM Advanced imaging cell analyzer (Chemometec, Denmark) measures cell viability.

[0254] result:

[0255] The results showed that, compared with alemtuzumab, PA1 induced stronger cytotoxic activity against all T-ALL and cutaneous T-cell lymphoma cell lines.

[0256] alemtuzumab showed low cytotoxicity against the tested tumor cell lines, but high cytotoxicity against PBMCs from healthy donors (see [link to article]). Figure 3 ).

[0257] Example 4: Apoptotic activity of the antibody combination of the present invention in different human cell lines

[0258] T-ALL cell lines (Jurkat, T1301, and HPB-ALL) and PTCL-NOS cell lines (300,000 cells per well) were exposed to a series of concentrations (from 10 µg / mL to 300 µg / mL) of the polyclonal antibody composition PA1 of the present invention.

[0259] In addition, T-ALL cell lines (Jurkat, T1301 and HPB-ALL), cutaneous T-cell lymphoma cell line (HUT78) and multiple myeloma cell line (300,000 cells per well) were exposed to a polyclonal antibody composition PA1 at a concentration of 250 µg / mL.

[0260] After incubation at 37°C for 20 hours, Annexin V-CF488A conjugate and Hoechst 33342 (final concentration: 10 µg / mL) were added, and incubation was continued at 37°C for 15 minutes. After washing, the cell pellet was resuspended in 100 µl Annexin V binding buffer supplemented with 10 µg / mL propidium iodide. NucleoCounter® NC-3000 was used. TM Advanced imaging cell analyzer (Chemometec, Denmark) analyzes apoptotic cells.

[0261] The same experimental protocol was also used to assess the apoptotic activity of PBMCs from healthy donors.

[0262] result:

[0263] The results showed that PA1 induced strong apoptotic activity in all tested T-ALL and PTCL-NOS cell lines, with the number of apoptotic cells ranging from 60% to 96% at a concentration of 100 µg / mL. These results demonstrate that PA1 specifically targets and kills tumor cells without damaging healthy PBMCs. No cross-reactivity with healthy PBMCs was observed at the highest tested concentration up to 300 µg / mL (see [link to study]). Figure 4 ).

[0264] As shown in Table 1 below, PA1 at a concentration of 250 µg / mL also induced strong and significant apoptosis in KMS-12-BM myeloma cells and HUT-78 cells.

[0265] Table 1

[0266]

[0267] Example 5: Comparison of the apoptosis activity of the antibody combination of the present invention and alenmab in different cell lines.

[0268] Apoptosis assays of the polyclonal antibody composition PA1 in different cell lines were compared with those of alemtuzumab (an anti-CD52 monoclonal antibody).

[0269] Apoptosis assays were performed on three T-ALL cell lines (HPB-ALL, Jurkat, and T1301). The polyclonal antibody composition PA1 and the monoclonal antibody (alemumab) were incubated at the same concentration of 30 µg / ml with the three different cell lines (300,000 tumor cells per well) at 37°C for 24 hours.

[0270] After incubation at 37°C for 24 hours, Annexin V-CF488A conjugate and Hoechst 33342 (final concentration: 10 µg / mL) were added, and incubation was continued at 37°C for 15 minutes. After washing, the cell pellet was resuspended in 100 µl Annexin V binding buffer supplemented with 10 µg / mL propidium iodide. NucleoCounter® NC-3000 was used. TM Advanced imaging cell analyzer (Chemometec, Denmark) analyzes apoptotic cells.

[0271] result:

[0272] In contrast to PA1, alemtuzumab showed weak apoptotic activity (less than 20%) against HPB-ALL and no apoptosis in T1301 and Jurkat cell lines (see [link]). Figure 5 ).

[0273] Example 6: Antibody-dependent cytotoxicity (ADCC) of the combination of antibodies of the present invention

[0274] According to MojoSort TM The Human NK Cell Isolation Kit Protocol (Biolegend, San Diego, California, USA) was used to isolate NK cells from PBMCs of healthy donors. On the day of assay, T-ALL cell lines (HPB-ALL) were labeled with carboxyfluorescein succinimide (CFSE) (Invitrogen, Waltham, Massachusetts, USA), incubated with a polyclonal antibody composition PA1 at concentrations of 10 µg / mL, 30 µg / mL, or 100 µg / mL, and co-cultured with human NK cells (E:T ratio 4:1) in RPMI containing 10% FCS for 16–24 hours. After 16–24 hours of incubation, propidium iodide was added, and the cells were isolated using a NucleoCounter® NC-3000 assay. TM Advanced imaging cell analyzer (ChemoMetec A / S, Allerrode, Denmark) analyzes cells (CFSE+ / IP+ cells).

[0275] result:

[0276] The results showed that the ADCC process is partially involved in PA1-induced cancer cell death (see...). Figure 6 ).

[0277] Example 7: Antibody-dependent phagocytosis (ADCP) of the antibody combination of the present invention

[0278] THP-1 cell lines were activated for 30 hours with PMA (phorbol myristate acetate, final concentration 20 ng / ml) to differentiate into macrophages, and then labeled with human anti-CD68 tagged with Alexa fluor 647. On the day of assay, T-ALL cell lines (HPB-ALL) were labeled with carboxyfluorescein succinimide (CFSE) (Invitrogen, Waltham, MA, USA) and incubated with a polyclonal antibody composition PA1 at concentrations of 10 µg / mL, 30 µg / mL, or 100 µg / mL, and co-cultured with human macrophages (E:T ratio 4:1) in RPMI containing 10% FCS for 16–24 hours. After 16–24 hours of incubation, propidium iodide was added, and the cells were cultured using NucleoCounter® NC-3000. TM Advanced imaging cell analyzer (ChemoMetec A / S, Allerrode, Denmark) analyzes cells (CFSE+ / IP+ cells).

[0279] result:

[0280] The results showed that ADCP process is involved in PA1-induced tumor cell death and exhibits a concentration-response relationship (see [link to relevant documentation]). Figure 7 ).

[0281] Example 8: Apoptosis pathways involved in combinations of antibodies of the present invention

[0282] To elucidate the apoptotic pathways involved in exposure to the combination of antibodies of the present invention, activation of caspase 8 (initiating the extrinsic apoptosis pathway) and caspase 9 (involved in the intrinsic apoptosis pathway) was investigated in tumor cell lines T1301, Jurkat, HUT-78, HPB-ALL, and KMS-12-BM. The cell lines were treated with the polyclonal antibody composition PA1 at 250 µg / ml at 37°C for 20 hours.

[0283] Twenty hours later, the activities of caspase 8 and caspase 9 were measured using specific fluorescent probes for caspase 8 or caspase 9, and the activity was determined using NucleoCounter® NC-3000. TM Advanced imaging cytometer (Chemometec, Denmark) analyzes fluorescence intensity.

[0284] result:

[0285] In all tested cell lines, both caspase 8 and caspase 9 were activated, indicating that PA1 simultaneously activates both exogenous and endogenous pathways (see [link to relevant documentation]). Figure 8 ).

[0286] Example 9: In vivo antitumor activity of the antibody combination of the present invention against T1301 xenograft.

[0287] A mouse model of T-ALL was obtained by subcutaneous injection of 3,000,000 T1301 cells on day 0.

[0288] This study included two groups of mice (n=10 in each group): a control group without treatment and a treatment group treated with the polyclonal antibody composition PA1. Treatment was initiated at the onset of tumor growth and lasted for a total of 28 days. Treatment consisted of intraperitoneal injections of 35 mg / kg PA1 twice weekly.

[0289] result:

[0290] PA1 demonstrated effectiveness in inhibiting tumor growth in vivo. On day 35, a 50% reduction in tumor size was observed in the PA1 treatment group compared to the mediator control group (see [link to relevant documentation]). Figure 10 ).

[0291] Furthermore, Table 2 below summarizes the EC obtained in the T1301 cell line using PA1 via in vitro CDC according to the protocols of Examples 1, 2, and 3. 20 EC 50 and EC Max .

[0292] Table 2

[0293]

[0294] Example 10: In vivo antitumor activity of the antibody combination of the present invention against Jurkat xenograft.

[0295] A xenograft mouse model of T-ALL was obtained by subcutaneously injecting 3,000,000 Jurkat cells in 50% Matrigel on day 0.

[0296] This study included two groups of mice (n=10 in each group): a control group without treatment and a treatment group treated with the polyclonal antibody composition PA1. Treatment was initiated at the onset of tumor growth and lasted for a total of 28 days. Treatment consisted of intraperitoneal injections of PA1 at 35 mg / kg twice weekly.

[0297] result:

[0298] PA1 demonstrated efficacy in inhibiting tumor growth in vivo. On day 40, compared to the mediator control group, a 72.3% reduction in tumor size was observed in the PA1 treatment group (see [link to relevant documentation]). Figure 12 ).

[0299] Furthermore, Table 3 below summarizes the EC obtained in the Jurkat cell line using PA1 via in vitro CDC according to the protocols of Examples 1, 2, and 3. 20 EC 50 and EC Max .

[0300] Table 3

[0301]

[0302] Example 11: Binding of the antibody combination of the present invention to PTCL tumors in patient biopsy tissue.

[0303] To evaluate the targeting and recognition capabilities of the antibody combination described in this invention in PTCL, the inventors evaluated the immunomarking of the polyclonal antibody composition PA1 of this invention on patient biopsy tissues. For this purpose, the inventors analyzed a tissue microarray (TMA) (Pantomics Inc., USA) consisting of 119 biopsy tissue cores from various PTCLs: 13 biopsy tissue cores from NK / T-cell lymphoma, 33 biopsy tissue cores from angioimmunoblastic T-cell lymphoma (AITL), 15 biopsy tissue cores from enteropathy-associated T-cell lymphoma (EATL), 46 biopsy tissue cores from PTCL-NOS, and 12 biopsy tissue cores from anaplastic large cell lymphoma (ALCL).

[0304] In summary, after dewaxing and antigen retrieval, TMA was incubated with the optimal concentration of PA1 at 2.5 µg / mL, followed by incubation with the secondary antibody-HRP conjugate, and then developed with ImmPACT VIP peroxidase substrate. The intensity and distribution of staining were graded using an internal grading system adapted from the standard grading system used in tissue cross-reactivity studies, employing ImageJ software. The grading system is provided in Table 4 below:

[0305] Table 4

[0306]

[0307] A case is considered positive if 10% or more of the tumor cells are stained with PA1.

[0308] result:

[0309] Surprisingly, PA1 identified 84.6% of NK / T PTCL biopsy tissues, 81.8% of AITL biopsy tissues, 93.3% of EATL biopsy tissues, 73.9% of NOS biopsy tissues, and 83.3% of ALCL biopsy tissues, as shown in Table 5 below:

[0310] Table 5

[0311]

[0312] In the biopsy tissues of the PTCL patients studied, the majority (>50%) had a score of 2, which can be considered high.

[0313] Micrographs show strong PA1 staining in NK / T-cell lymphoma, AITL, EATL, and NOS PTCL (all scores 2). Micrographs also show unstained PTCL within the same TMA (score 0).

[0314] Example 12: In vitro CDC study of the antibody combination of the present invention in different human solid tumor cell lines

[0315] Different solid cancer cell lines were exposed to a series of antitumor polyclonal antibody compositions PA1 (n=3) in the presence of rabbit complement (final dilution 1 / 3). The concentrations ranged from 12.5 µg / mL to 800 µg / mL.

[0316] The solid cancer cell lines tested are as follows:

[0317] - The A549 cell line represents non-small cell lung cancer.

[0318] The Hep-G2 cell line represents hepatocellular carcinoma.

[0319] -HCT-116 cell line represents colorectal cancer.

[0320] The MDA-MB-231 cell line represents triple-negative breast cancer.

[0321] -LNCAP cell line represents prostate cancer.

[0322] -SK-MEL-30 cell line represents melanoma.

[0323] After incubating at 37°C for 1 hour, NucleoCounter® NC-3000 was used. TM Advanced imaging cell analyzer (Chemometec, Denmark) measures cell viability.

[0324] result:

[0325] PA1 induced strong cytotoxic activity against human solid tumor cell lines (ranging from 45.2% to 90.7%) (see [link to relevant documentation]). Figure 13 ).

[0326] Example 13: In vitro apoptosis activity of the antibody combination of the present invention in different human solid tumor cell lines

[0327] Different solid cancer cell lines (300,000 cells per well) were exposed to a polyclonal antibody composition PA1 at a concentration of 250 µg / mL.

[0328] The solid cancer cell lines tested are as follows:

[0329] - The A549 cell line represents non-small cell lung cancer.

[0330] The Hep-G2 cell line represents hepatocellular carcinoma.

[0331] -HCT-116 cell line represents colorectal cancer.

[0332] The MDA-MB-231 cell line represents triple-negative breast cancer.

[0333] -LNCaP cell line represents prostate cancer.

[0334] After incubation at 37°C for 20 hours, Annexin V-CF488A conjugate and Hoechst 33342 (final concentration: 10 µg / mL) were added, and incubation was continued at 37°C for 15 minutes. After washing, the cell pellet was resuspended in 100 µl Annexin V binding buffer supplemented with 10 µg / mL propidium iodide. NucleoCounter® NC-3000 was used. TM Advanced imaging cell analyzer (Chemometec, Denmark) analyzes apoptotic cells.

[0335] result:

[0336] PA1 induced apoptosis in the tested cancer cell lines. Results for each cell line are provided in Table 6 below:

[0337] Table 6

[0338]

[0339] Example 14: The whole cells of the antibody combination of the present invention after incubation for 24 hours in different human solid tumor cell lines Cytotoxicity

[0340] Different solid cancer cell lines were exposed to the polyclonal antibody composition PA1 in the presence of rabbit complement (final dilution 1 / 3) at concentrations ranging from 12.5 µg / mL to 800 µg / mL.

[0341] The solid cancer cell lines tested are as follows:

[0342] - The A549 cell line represents non-small cell lung cancer.

[0343] The Hep-G2 cell line represents hepatocellular carcinoma.

[0344] -HCT-116 cell line represents colorectal cancer.

[0345] The MDA-MB-231 cell line represents triple-negative breast cancer.

[0346] -LNCAP cell line represents prostate cancer.

[0347] -SK-MEL-30 cell line represents melanoma.

[0348] After incubation at 37°C for 24 hours, NucleoCounter® NC-3000 was used. TM Advanced imaging cell analyzer (Chemometec, Denmark) measures cell viability.

[0349] result:

[0350] PA1 induced strong cytotoxicity (ranging from approximately 50% to 95%) in human solid tumor cell lines (see [link]). Figure 14 ).

[0351] Example 15: Target of the antibody combination of the present invention

[0352] Using HuProt TM The array (CDI Labs, USA) was used for the assay of the polyclonal antibody composition of the present invention, named PA1. After blocking, the array was probed with PA1 (1 µg / ml) at room temperature for 1 hour. The array was then washed three times with TBST for 10 minutes each time and probed with Alexa647-anti-pig IgG secondary antibody under CDI Labs-optimized signal detection conditions.

[0353] The Z-score is the average Z-score of a given protein at replicate points (for each protein in HuProt). TM (Printed repeatedly on the array). The Z-score for each point on the given array is calculated using the following algorithm: Z = [F635 – F635(avg)] / F635(std). F635(avg) and F635(std) are the mean and standard deviation of the F635 values ​​for all points on the array, respectively.

[0354] result:

[0355] The polyclonal antibody composition was observed to bind to all six protein targets.

[0356] Example 16: In vitro cytotoxicity assay

[0357] Different liquid cancer cell lines were exposed to the polyclonal antibody composition PA1 in the presence of rabbit complement (final dilution 1 / 3) at concentrations ranging from 12.5 µg / mL to 800 µg / mL.

[0358] The liquid cancer cell lines tested are as follows:

[0359] HPB-ALL, Jurkat, and T1301 cell lines represent T-ALL.

[0360] -HUT-78 cell line represents cutaneous T-cell lymphoma (Cezari syndrome), and

[0361] -KARPAS 299 cell line represents ALCL.

[0362] After incubation at 37°C for 24 hours, cell viability was studied using the CellTiter-Glo® cell viability assay. Cell viability was quantified using the CellTiter-Glo® One Solution cell viability assay (Promega G8462). After cell incubation, the CellTiter-Glo® One Solution assay reagent was brought to ambient temperature. Then, 100 µl of the CellTiter-Glo® One Solution assay reagent was added to each well. The plate was shaken for 2 minutes to induce cell lysis, and incubated for 20 minutes before reading the luminescence (LU) value using a GloMax microplate reader (Promega).

[0363] result:

[0364] The results showed that PA1 induced strong cell death after 24 hours of incubation, even in the HUT78 cell line, which is described as a cell line that does not express the CD99 membrane protein (see [link to study]). Figure 18 ).

[0365] Example 17: In vivo efficacy comparison with CHOP in a rat T-cell lymphoma / leukemia model

[0366] 15x10 in 50% matrigel 6 One T1301 cell (final volume 500 µl) was subcutaneously injected into the left abdomen of SRG immunodeficient rats. Tumor growth was monitored twice weekly using calipers. Once the tumor reached 500-1000 mm... 3 Treatment begins, and according to the above protocol, it consists of one cycle of CHOP chemotherapy or PA1 treatment twice a week.

[0367] Design: 3 groups, n=7 rats in each group

[0368] Group 1: Control group without treatment (control group)

[0369] Group 2: PA1 treatment, twice weekly: 40 mg / kg (PA1 group); and

[0370] Group 3: CHOP treatment (1 cycle): 1 cycle = D0: cyclophosphamide 37.5 mg / kg, hydroxydaunorubicin 2.5 mg / kg, and vincristine 0.07 mg / kg, and D0, D1, D2, D3, D4, D5: prednisone 1.47 mg / kg (CHOP group).

[0371] result:

[0372] The results showed that PA1 administration completely halted tumor growth during treatment. No tumor escape was observed with PA1 treatment.

[0373] Five days after the end of the CHOP chemotherapy cycle, D10, tumor growth slowed in the CHOP group, with a 40% reduction observed at day 21. Tumor progression occurred after day 21, and appeared to be similar to the tumor growth progression in the control group.

[0374] PA1 demonstrated high efficacy and good tolerability in SRG T1301 xenografting, while only minor and non-durable effects were observed with standard care (CHOP) (see [link to relevant documentation]). Figure 16 and Figure 17 ).

Claims

1. A combination of antibodies or antigen-binding fragments thereof, said combination for the treatment of cancer in human subjects of need, said combination comprising: - At least a first antibody or its antigen-binding fragment thereof, wherein the first antibody or its antigen-binding fragment specifically binds to the FASN (FasN fragment); - At least a second antibody or its antigen-binding fragment, wherein the second antibody or its antigen-binding fragment specifically binds to CD99 (differentiation cluster 99); and - At least a third antibody or its antigen-binding fragment thereof, said third antibody or its antigen-binding fragment specifically binding to an antigen selected from the group consisting of GPI (glucose-6-phosphate isomerase), SLC3A2 (solute carrier family 3 member 2), GART (glycamide ribonucleotide formyltransferase) and CKAP4 (cytoskeleton-associated protein 4), particularly the group consisting of GPI and GART.

2. The combination used according to claim 1, wherein the combination further comprises at least a fourth antibody or antigen-binding fragment thereof, the fourth antibody or antigen-binding fragment thereof specifically binding to an antigen selected from the group consisting of GPI, SLC3A2, GART and CKAP4, particularly the group consisting of GPI and GART, the fourth antibody or antigen-binding fragment thereof being different from the third antibody or antigen-binding fragment thereof.

3. The combination used for the purpose described in claim 1 or 2, wherein the combination comprises at least: -At least a first antibody or its antigen-binding fragment thereof, wherein the first antibody or its antigen-binding fragment specifically binds to FASN; -At least a second antibody or its antigen-binding fragment thereof, wherein the second antibody or its antigen-binding fragment specifically binds to CD99; - At least a third antibody or its antigen-binding fragment, said third antibody or its antigen-binding fragment specifically binding to GPI; and - At least a fourth antibody or its antigen-binding fragment, said fourth antibody or its antigen-binding fragment specifically binds to GART.

4. The combination used according to any one of claims 1 to 3, wherein the antigen-binding fragment of the antibody present in the combination is independently Fv, Fab, F(ab')2, Fab', dsFv, (dsFv)2, scFv, sc(Fv)2 or a double-chain antibody.

5. The combination of uses according to any one of claims 1 to 4, wherein said cancer is selected from the group consisting of: myeloma; melanoma; skin cancer; breast cancer; brain tumor, bladder cancer, cervical cancer, prostate cancer; primary and metastatic colorectal cancer, particularly colon cancer; mesothelioma; lung cancer, particularly non-small cell lung cancer; liver cancer, particularly hepatocellular carcinoma or bile duct cancer; primary and metastatic pancreatic cancer; kidney cancer; soft tissue sarcoma; thyroid cancer; lymphoma, including Hodgkin lymphoma and non-Hodgkin lymphoma, particularly B-cell lymphoma or T-cell lymphoma, more particularly T-cell lymphoma; gastric cancer; head and neck cancer; ovarian cancer; sarcoma; acute or chronic leukemia, particularly T-cell leukemia or myeloid leukemia; osteosarcoma; anal cancer; testicular cancer; uterine cancer; thyroid cancer; central nervous system cancer; gastrointestinal stromal carcinoma; epidermal cancer; and esophageal cancer.

6. The combination used according to any one of claims 1 to 5, wherein the cancer is: - Lymphoma, especially those selected from the group consisting of the following: (i) Non-Hodgkin lymphoma selected from the group consisting of the following: - B-cell lymphomas, particularly those selected from the group consisting of: diffuse large B-cell lymphoma, follicular lymphoma, small lymphocytic lymphoma (or chronic lymphocytic leukemia), mantle cell lymphoma, marginal zone B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma (or Waldenström macroglobulinemia), hairy cell leukemia, and primary central nervous system lymphoma; and -T-cell lymphomas, particularly those selected from the group consisting of: precursor T-lymphoblastic lymphoma (or precursor T-lymphoblastic leukemia), peripheral T-cell lymphoma, and cutaneous T-cell lymphoma; and (ii) Hodgkin lymphoma selected from the following groups: tuberous sclerosis Hodgkin lymphoma, mixed cellularity Hodgkin lymphoma, lymphocytic depletion Hodgkin lymphoma, and lymphocyte-rich Hodgkin lymphoma; or - Leukemia, particularly those selected from the following groups: acute myeloid leukemia (AML), chronic myeloid leukemia (CML), and histiocytic leukemia.

7. The combination of uses according to any one of claims 1 to 6, wherein the cancer is selected from the group consisting of: -T-cell acute lymphoblastic leukemia (T-ALL); - Peripheral T-cell lymphoma (PTCL), especially PTCL selected from the following groups: PTCL nonspecific type (PTCL-NOS), enteropathy-associated T-cell lymphoma (EATL), monomorphic epithelial-friendly intestinal T-cell lymphoma (MEITL), anaplastic large cell lymphoma (ALCL) and angioimmunoblastic T-cell lymphoma (AITL), extranodal NK / T-cell lymphoma nasal type (ENKL), hepatosplenic γδ T-cell lymphoma (HSGDTCL), intestinal T-cell lymphoma (ITCL), mycosis fungoides and Cezari syndrome; - Cutaneous T-cell lymphoma (CTCL); and -T-cell lymphoblastic lymphoma (T-LBL).

8. The combination used according to any one of claims 1 to 7, wherein the cancer expresses at least one antigen selected from the group consisting of FASN, CD99, GPI, SLC3A2, GART and CKAP4, particularly the group consisting of GPI and GART.

9. The combination used according to any one of claims 1 to 8, wherein at least one antibody in the combination, and in particular all antibodies in the combination, lacks at least one antigenic determinant selected from (i) N-hydroxyacetylneuraminic acid (Neu5Gc) and (ii) α-1,3-galactose, and in particular lacks two antigenic determinants: N-hydroxyacetylneuraminic acid (Neu5Gc) and α-1,3-galactose.

10. The combination used for any one of claims 1 to 9, wherein the combination is a combination of monoclonal antibodies or a polyclonal antibody composition.

11. The combination used according to claim 10, wherein the polyclonal antibody composition can be obtained by immunizing animals with CD3+, CD8+, TCR γ / δ- and CD34- human tumor T cells.

12. The combination used according to claim 11, wherein the animal is a pig, and particularly a pig lacking at least one gene selected from the group comprising (i) a gene encoding a functional cytidine-5'-monophosphate N-acetylneuraminic acid hydrolase (CMAH) and (ii) a gene encoding a functional α-(1,3)-galactosyltransferase, and more particularly a pig lacking both (i) a gene encoding a functional cytidine-5'-monophosphate N-acetylneuraminic acid hydrolase (CMAH) and (ii) a gene encoding a functional α-(1,3)-galactosyltransferase.

13. The combination used according to any one of claims 1 to 12, wherein the combination comprises a pharmaceutical composition, the pharmaceutical composition further comprising a pharmaceutically acceptable carrier.

14. The combination used according to claim 13, wherein the pharmaceutical composition further comprises at least one additional anticancer drug, said additional anticancer drug being different from the antibody in the combination of antibodies as defined in any one of claims 1 to 4 and 8 to 11.

15. The combination used according to claim 14, wherein the at least one additional anticancer drug is selected from the group consisting of monoclonal antibodies, particularly from the group consisting of: anti-CD19, anti-CD20, anti-CD30, anti-CD137, anti-CTLA4, anti-TIM-3, anti-B7-H3, anti-CD123, anti-CD134, anti-CD154, anti-LAG-3, anti-CD227, anti-BTNA3, anti-CD39, anti-CD73, anti-CD115, anti-CD47, anti-SIRPα, anti-SIRPγ, anti-CD28, anti-NCR, anti-NKp46, anti-NKp30, anti-NKp44, anti-NKG2D, anti-PD1, anti-PDL1, moglizazumab, oxotuzumab, velpotuzumab, yttrium Y90-tiemozumab, motuzumab, and anti-DNAM-1 monoclonal antibody.

16. The combination used according to any one of claims 13 to 15, wherein the pharmaceutical composition further comprises at least one chemotherapy treatment, particularly chemotherapy treatment selected from the group consisting of: a chemotherapy regimen consisting of cyclophosphamide, hydroxydaunorubicin, vincristine, and prednisone (CHOP); a chemotherapy regimen consisting of cyclophosphamide, hydroxydaunorubicin, vincristine, etoposide, and prednisone (CHOEP); an HDAC inhibitor; ibrutinib, acalabrutinib, zanubrutinib, curapannisin, and veneclade.

17. The combination used for any one of the uses described in claims 1 to 16, wherein the combination is used to provide antitumor activity selected from the group consisting of complement-dependent cytotoxicity (CDC), complement-dependent cytotoxicity (ADCC) from killer cells, apoptotic activity and / or complement-dependent cytotoxicity (ADCP) from phagocytes, and particularly for providing antitumor activity consisting of complement-dependent cytotoxicity (CDC), complement-dependent cytotoxicity (ADCC) from killer cells, apoptotic activity and complement-dependent cytotoxicity (ADCP) from phagocytes.

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