Combination of inhibitors of il-1beta, CCR5 or CXCR2 cytokine receptors for use in the treatment of metastasis

Abstract

The present invention relates to a combination of at least two inhibitors of cytokine receptors or to a pharmaceutical composition comprising same, for use in the prevention and treatment of metastasis. The composition comprises an inhibitor of the IL-1β cytokine receptor and / or an inhibitor of the CCR5 cytokine receptor and / or an inhibitor of the CXCR2 cytokine receptor. The pharmaceutical composition comprises a combination of at least two inhibitors of cytokine receptors, such as an inhibitor of the IL-1β cytokine receptor and / or an inhibitor of the CCR5 cytokine receptor and / or an inhibitor of the CXCR2 cytokine receptor.

Description

[0001] DESCRIPTION

[0002] CYTOKINE RECEPTOR INHIBITORS FOR USE IN THE PREVENTION AND TREATMENT OF METASTASIS

[0003] TECHNICAL SECTOR

[0004] The present invention relates to the prevention and treatment of metastasis. In particular, the present invention relates to the prevention and treatment of metastasis with cytokine receptor inhibitors.

[0005] BACKGROUND OF THE INVENTION

[0006] Metastasis is the spread of cancer cells from the site where the cancer first formed to another part of the body. Metastasis occurs when cancer cells break away from the original (primary) tumor, travel through the body via the bloodstream or lymphatic system, and form a new tumor in other organs or tissues. This new tumor can, in turn, generate further metastasis in a different organ or tissue.

[0007] The spread of cancer cells through metastasis is associated with cellular mechanisms detailed below. These cellular mechanisms are, in turn, related to signaling pathways.

[0008] Cell migration is the first step in metastasis, allowing cancer cells to move from the primary tumor to other tissues. This process is crucial for cancer cells to invade adjacent tissues and eventually enter the bloodstream or lymphatic system, facilitating their spread.

[0009] Invasion is a critical step in metastasis, where cancer cells penetrate and degrade the extracellular matrix. Another equally critical step in metastasis is transvasation (also called transendothelial migration), the process by which tumor cells cross the walls of blood or lymphatic vessels to enter the circulation. This step is essential for cancer cells to travel through the bloodstream or lymphatic system and colonize distant organs.

[0010] Another important step in metastasis is the epithelial-mesenchymal transition, a phenotypic change in which epithelial cells acquire mesenchymal characteristics, including increased migratory and invasive capabilities. This change is crucial for metastasis because it allows tumor cells to detach from the primary tumor and resist apoptosis.

[0011] Tumors with a risk of generating secondary tumors in other organs or tissues through metastasis include, among others, triple-negative breast cancer, prostate cancer, lung cancer, and osteosarcoma.

[0012] A high percentage of deaths in cancer patients are due to cancer metastasis, so there is an urgent need to identify the molecular and cellular mechanisms of metastasis in order to design new therapeutic strategies to combat it.

[0013] Furthermore, some patients with cancers at risk of metastasis do not respond well to standard treatments such as chemotherapy and have an unfavorable outcome, so there is an urgent need to design new, more effective therapeutic strategies for these patients.

[0014] DESCRIPTION OF THE INVENTION

[0015] The technical problem to be solved is how to prevent or treat metastasis effectively.

[0016] Additionally, the technical problem to be solved is to prevent or treat metastasis effectively in subjects who do not respond to chemotherapy.

[0017] The present invention solves this technical problem by providing a combination of at least two cytokine receptor inhibitors or a composition comprising them.

[0018] The inventors have studied the molecular mechanisms underlying metastasis and found that cytokines play a crucial role at every stage of the process. Communication between tumor cells and tumor-associated macrophages facilitates the survival and proliferation of cancer cells and also promotes their ability to migrate, invade, and spread throughout the body. Therapeutic interventions with cytokine receptor inhibitors block these metastatic processes. Cytokine receptor inhibitors can block cytokine-mediated signaling, preventing the activation of tumor cell migration, invasion, and transvasation. These strategies offer a promising approach to preventing metastasis.Through orthological, immunological, gene expression, and cell co-culture studies, the inventors have found that patients who do not respond to chemotherapy have higher levels of immunosuppression. Evasion of response to chemotherapy is a hallmark of tumor progression and is triggered by mechanisms that regulate immune checkpoints in both immune and tumor cells.

[0019] First, the tumor microenvironment is composed of immunosuppressive myeloid cells, which are of two types: myeloid-derived suppressor cells and tumor-associated macrophages. The inventors studied the conditioned medium obtained from the co-culture of tumor cells with non-polarized M0 macrophages. In human tissues, three main macrophage subpopulations coexist: non-polarized M0 macrophages and polarized M1 (pro-inflammatory macrophages) and M2 (anti-inflammatory macrophages) macrophages. Macrophage polarization is a process by which macrophages adopt different functions in response to signals from their microenvironment.

[0020] The conditioned medium obtained from the co-culture of tumor cells with non-polarized M0 macrophages significantly increases cell migration, invasion, and transendothelial migration of triple-negative breast cancer cells, the first steps in the metastasis process. Through analysis of the conditioned medium, the inventors have identified a cocktail of factors responsible for the increased aggressiveness and metastasis of the tumor cells. Using expression arrays and subsequent validation by ELISA, the inventors have found that the cocktail contains the cytokines CCL3, CCL4, CXCL2, and IL-1β. The present invention proposes to pharmacologically inhibit the receptors of these cytokines, thereby significantly decreasing cell migration, as well as invasion and transendothelial migration.

[0021] Additionally, the inventors have studied the expression of epithelial-mesenchymal transition (EMT) markers using digital PCR. Treatment with cytokine inhibitors decreases the expression of EMT markers such as CDH1, CDH2, TWIST1, and MMP9. Therefore, treatment with cytokine inhibitors blocks not only the initiation of metastasis but also the EMT, which is also involved in drug resistance and cell migration. Interleukin-1 p (IL-1 p) is a cytokine that promotes EMT and tumor invasion. IL-1 is one of two forms of IL-1; the other form is IL-1a. For the purposes of the present invention, the terms “IL-1” and “IL-1 p” may be used interchangeably.

[0022] IL-1 p cytokine receptor inhibitors block the binding of IL-1 p to its receptor and partially reverse the epithelial-mesenchymal transition and reduce the invasive capacity of tumor cells.

[0023] CCR5 is a cytokine receptor found primarily on immune system cells, such as T lymphocytes and macrophages. CCR5 is also present on some types of tumor cells. CCR5 inhibitors block the binding of the cytokines CCL3 and CCL4 to CCR5 and reduce cytokine-induced tumor cell migration and invasion by interfering with cross-communication between tumor-associated macrophages and tumor cells.

[0024] CXCR2 is a cytokine receptor that plays a crucial role in the attraction and activation of neutrophils and other immune cells that contribute to the pro-metastatic tumor microenvironment. CXCR2 inhibitors block the binding of the cytokine CXCL2 to CXCR2 and reduce inflammation and the pro-tumor immune response, decreasing cell migration and invasion.

[0025] EZH2 is an epigenetic regulatory enzyme, the enhancer of Zeste Homolog 2 (EZH2). EZH2 is a histone lysine methyltransferase that methylates lysine 27 of histone H3. EZH2 inhibitors reduce the expression of pro-metastatic genes and reverse changes associated with the epithelial-mesenchymal transition.

[0026] For the purposes of the present invention, “IL-6” refers to interleukin 6.

[0027] For the purposes of the present invention, “PD-1” refers to programmed cell death protein 1 (PD-1), a surface receptor expressed on the surface of T cells and B cells. PD-1 is an immunoregulatory receptor and an immune checkpoint. PD-1 inhibitors activate the immune system and are used to treat certain types of cancer. For the purposes of the present invention, “PD-L1” refers to programmed cell death ligand 1 (PD-L1), a protein that binds to the PD-1 receptor. PD-L1 inhibitors block PD-L1's binding to PD-1 and activate the immune system.

[0028] The present invention provides a combination of at least two cytokine receptor inhibitors or a pharmaceutical composition comprising them, for use in the prevention or treatment of metastasis, wherein the cytokines consist of IL-1p, CCR5 or CXCR2.

[0029] Additionally, the present invention relates to the use of a combination of at least two cytokine receptor inhibitors or a pharmaceutical composition comprising them, wherein the cytokines consist of IL-1p, CCR5, or CXCR2, to prepare a medicament useful in the prevention or treatment of metastasis. Preferably, the combination or composition for preparing the medicament comprises an IL-1p cytokine receptor inhibitor, a CCR5 cytokine receptor inhibitor, and a CXCR2 cytokine receptor inhibitor.

[0030] Additionally, the present invention relates to a method for preventing or treating metastasis in a subject, wherein the subject is administered a combination of at least two cytokine receptor inhibitors or a pharmaceutical composition comprising them, wherein the cytokines consist of IL-1p, CCR5, or CXCR2. Preferably, the combination or composition comprises an IL-1p cytokine receptor inhibitor, a CCR5 cytokine receptor inhibitor, and a CXCR2 cytokine receptor inhibitor.

[0031] In another preferred embodiment of the combination or composition for use of the invention, at least one of the two inhibitors present in the combination is an inhibitor of the cytokine I L-1 p receptor.

[0032] In a more preferred embodiment of the combination or composition for use of the invention, the IL-1 cytokine receptor inhibitor is selected from the group consisting of: anakinra, canakinumab, rilonacept (also known as “IL-1 Trap”), belsomra, gevokizumab (also known as “Xoma-052”), lutikizumab, GSK 1070806, dapansutrile, and zerumbona, or combinations thereof. More preferably, the IL-1 cytokine receptor inhibitor is anakinra. Anakinra has been used as a medicament to treat autoimmune diseases, such as rheumatoid arthritis and familial Mediterranean fever. In another preferred embodiment of the combination or composition for use of the invention, at least one of the two inhibitors present in the combination is a CCR5 cytokine receptor inhibitor.

[0033] In a more preferred embodiment of the combination or composition for use of the invention, the CCR5 cytokine receptor inhibitor is selected from the group consisting of: maraviroc, Met-CCL3, Met-CCL3 / CCL4, plerixafor (also referred to as “AMD3100”), cucurbitacin I, leronlimab (also referred to as “PRO 140”), naltrexone, dihydro-7H-benzo[7]annulene-6-carbonyl]amino]phenyl]methyl]-(oxan-4-yl)azanium (also referred to as “TAK-779”) and

[0034] 1 [2- [4- (trif I or orom ethyl)fe ni I ]et¡ l]es pyro[1 H-3, 1 -benzoxazine-4,4'-p¡peridin]-2-one

[0035] (also known as “RS 102895”), or combinations thereof. Most preferably, the CCR5 inhibitor is maraviroc. Maraviroc has been used as an antiretroviral in the treatment of HIV, blocking the virus from entering CD4+ cells.

[0036] In another preferred embodiment of the combination or composition for use of the invention, at least one of the two inhibitors present in the combination is an inhibitor of the CXCR2 cytokine receptor.

[0037] In a more preferred embodiment of the combination or composition for use of the invention, the CXCR2 cytokine receptor inhibitor is selected from the group consisting of navarixin (also known as “SCH-527123”), danirixin, elubrixin (also known as “SB-656933”), and reparixin, or combinations thereof. More preferably, the CXCR2 inhibitor is navarixin. Navarixin has been used in the treatment of respiratory diseases, particularly obstructive diseases such as asthma and chronic obstructive pulmonary disease (COPD).

[0038] In a preferred embodiment, the combination or composition of the invention for use in the treatment of metastatic cancers or tumors, particularly those unresponsive to other chemotherapy treatments, and more specifically for use in the treatment of metastatic triple-negative breast cancer, comprises at least one inhibitor of the cytokine I L-1p receptor, at least one inhibitor of the cytokine receptor CCR5, and at least one inhibitor of the cytokine receptor CXCR2. The inventors discovered that several cytokines independently affect metastasis, and therefore combining inhibitors against each of them is more efficient at blocking metastasis than using each inhibitor separately.

[0039] The combination of IL-1β cytokine receptor inhibitors, CCR5 cytokine receptor inhibitors, and CXCR2 cytokine receptor inhibitors blocks multiple signaling pathways involved in metastasis: (i) inhibition of migration and invasion: CCR5 and CXCR2 inhibitors reduce the migration and invasion of tumor cells, limiting their ability to spread from the primary tumor; (ii) reversal of the epithelial-mesenchymal transition: IL-1β cytokine receptor inhibitors reverse the epithelial-mesenchymal transition, making tumor cells less invasive and more susceptible to apoptosis; and (iii) reduction of cross-communication: together, these cytokine receptor inhibitors disrupt cross-communication between tumor cells and tumor-associated macrophages, weakening the pro-metastatic tumor microenvironment and reducing cancer progression.Blocking multiple signaling pathways involved in metastasis prevents cancer cells from escaping. Blocking escape pathways related to cell migration, cell invasion, and transvasation prevents cancer cells from initiating metastasis and increases the effectiveness of treatment for cancers and tumors that metastasize.

[0040] In a further more preferred embodiment, the combination or composition for use of the invention comprises at least one IL-1β cytokine receptor inhibitor, at least one CCR5 cytokine receptor inhibitor, and at least one CXCR2 cytokine receptor inhibitor. More preferably, the IL-1β cytokine receptor inhibitor is anakinra, the CCR5 cytokine receptor inhibitor is maraviroc, and the CXCR2 cytokine receptor inhibitor is navahxin.

[0041] In another preferred embodiment of the invention, the combination or composition further comprises at least one EZH2 inhibitor. Preferably, the EZH2 inhibitor is selected from the group consisting of tazemetostat, (S)-1-(sec-butyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-methyl-6-(6-(piperazin-1-yl)pyridin-3-yl)-1H-indole-4-carboxamide (also referred to as “GSK-126”), lirametostat (also referred to as “CPI-1205”), valemetostat (also referred to as “DS-3201”), EZM-002 (also referred to as “PF-06821635”), mevrometostat (also referred to as “PF-06821497”), N-[(6-methyl-2-oxo-4-propyl-1H-pyridin-3-yl)methyl]-1 -propan-2-yl-6-[6-(4-propan-2-ylpiperazin-1-yl)pyridin-3-yl]indazol-4-carboxamide (also referred to as “UNC-1999”), 3-[ethyl(tetrahydro-2H-pyran-4-yl)amino]-N-[(2,3,5,6,7,8-hexahydro-1-methyl-3-oxo-4-isoquinolinyl)methyl]-2-methyl-5-[6-(4-methyl-1-piperazinyl)-3-pyridinyl]-benzamide (also referred to as “ZLD1039”) and OR-S1, or combinations thereof.

[0042] In another preferred embodiment of the invention, the combination or composition further comprises at least one IL-6 cytokine receptor inhibitor. Preferably, the IL-6 cytokine receptor inhibitor is selected from the group consisting of: tozilizumab, siltuximab, sarilumab, olokizumab, clazakizumab, and levilimab, or combinations thereof.

[0043] In another preferred embodiment of the invention, the combination or composition further comprises at least one PD-1 receptor inhibitor. Preferably, the PD-1 receptor inhibitor is selected from the group consisting of: nivolumab, pembrolizumab, cemiplimab, dostarlimab, toripalimab, and tislelizumab, or combinations thereof.

[0044] In another preferred embodiment of the invention, the combination or composition further comprises at least one PD-L1 inhibitor. Preferably, the PD-L1 inhibitor is selected from the group consisting of: atezolizumab, durvalumab, avelumab, and envafolimab, or combinations thereof.

[0045] In a further preferred embodiment of the invention, the combination or composition for use in the treatment of cancers or metastatic tumors comprises at least one inhibitor of the cytokine I L-1 p receptor, at least one inhibitor of the cytokine receptor CCR5, at least one inhibitor of the cytokine receptor CXCR2, and at least one inhibitor of EZH2.

[0046] In one embodiment of the invention, the combination or composition is for use in the treatment of aggressive cancers that progress with metastasis, such as triple-negative breast cancer, prostate cancer, lung cancer, and osteosarcoma.

[0047] Therefore, in a preferred embodiment of the combination or composition for use of the invention, the metastasis originated from a tumor selected from the group consisting of: breast cancer, prostate cancer, lung cancer, and osteosarcoma. It has been described that surgery to remove a tumor with a curative objective of eliminating and reducing the tumor mass can, paradoxically, lead to an increase in metastases. Similarly, a biopsy can also lead to an increase in metastases.

[0048] Therefore, in a preferred embodiment of the invention, the combination or composition is for use in a subject who has undergone a tumor resection or biopsy.

[0049] In a preferred embodiment of the invention, the combination or composition is for use in a cancer or tumor that does not respond to chemotherapy.

[0050] In an even more preferred embodiment of the combination or composition for use of the invention, the metastasis originated in a triple-negative breast cancer.

[0051] Triple-negative breast cancer is defined by the absence of estrogen receptor, progesterone receptor, and HER2 expression. Compared to other breast cancer subtypes, triple-negative breast cancer is more aggressive, with a greater propensity for metastasis and recurrence. Triple-negative breast cancer is also highly heterogeneous, with diverse molecular and genetic alterations that result in varied responses to neoadjuvant chemotherapy, which is administered as a first step to reduce tumor size before the main treatment, which usually consists of surgery.

[0052] The progression of triple-negative breast cancer is closely linked to immune evasion and immunotolerance mechanisms. Furthermore, tissue-resident macrophages, the most abundant stromal cells in early-stage triple-negative breast cancer before angiogenesis, play a crucial role in tumor progression.

[0053] In triple-negative breast cancer, there is a high rate of complete positive response to neoadjuvant chemotherapy. However, refractory patients or those with a poor response often have an unfavorable outcome, making it urgently necessary to develop new, more effective treatments than neoadjuvant chemotherapy for metastatic triple-negative breast cancer. In a further preferred embodiment of the invention, the combination or composition is for use in the treatment of metastatic triple-negative breast cancer in a subject who is unresponsive to chemotherapy.

[0054] In a more preferred embodiment, the composition of the invention further comprises at least one pharmaceutically acceptable vehicle or excipient.

[0055] The present invention also provides a pharmaceutical composition comprising at least two cytokine receptor inhibitors, wherein the cytokines consist of IL-1P, CCR5, or CXCR2.

[0056] The pharmaceutical composition of the invention can be administered by any route of administration (for example, oral, sublingual, perioral, parenteral, intraperitoneal, intramuscular, intranasal, intravenous, intra-arterial, transdermal, subcutaneous, topical, etc.) for which the composition will be formulated in the pharmaceutical form appropriate to the chosen route of administration.

[0057] The pharmaceutical composition of the invention can be formulated to provide controlled release of the active ingredients, such as sustained or prolonged release, in accordance with conventional techniques in pharmaceutical practice.

[0058] In a preferred embodiment of the pharmaceutical composition of the invention, at least one of the two inhibitors present in the composition is a cytokine I L-1 p receptor inhibitor. Preferably, the cytokine I L-1 p receptor inhibitor is selected from the group consisting of: anakinra, canakinumab, rilonacept, belsomra, gevokizumab, lutikizumab, GSK 1070806, dapansutrile, and zerumbona, or combinations thereof. More preferably, the cytokine I L-1 p receptor inhibitor is anakinra.

[0059] In another more preferred embodiment of the pharmaceutical composition of the invention, at least one of the two inhibitors present in the combination is a CCR5 cytokine receptor inhibitor. Preferably, the CCR5 cytokine receptor inhibitor is selected from the group consisting of: maraviroc, Met-CCL3, Met-CCL3 / CCL4, plerixafor, cucurbitacin I, leronlimab, naltrexone, dimethyl-[[4-[[3-(4-methylphenyl)-8,9-dihydro-7H-benzo[7]annulene-6-carbonyl]amino]phenyl]methyl]-(oxan-4-yl)azanium and

[0060] 1 [2- [4- (trif I u orom eti l)fe ni I ]et¡ l]es pi ro[1 H-3, 1 -benzoxazine-4,4'-piperidine]-2-one, or combinations thereof. More preferably, the CCR5 cytokine receptor inhibitor is maraviroc.

[0061] In another more preferred embodiment of the pharmaceutical composition of the invention, at least one of the two inhibitors present in the combination is a CXCR2 cytokine receptor inhibitor. Preferably, the CXCR2 cytokine receptor inhibitor is selected from the group consisting of navarixin, danirixin, elubrixin, and reparixin, or combinations thereof. More preferably, the CXCR2 cytokine receptor inhibitor is navarixin.

[0062] In another more preferred embodiment, the pharmaceutical composition of the invention comprises at least one IL-1β cytokine receptor inhibitor, at least one CCR5 cytokine receptor inhibitor, and at least one CXCR2 cytokine receptor inhibitor. More preferably, the IL-1β cytokine receptor inhibitor is anakinra, the CCR5 cytokine receptor inhibitor is maraviroc, and the CXCR2 cytokine receptor inhibitor is navarixin.

[0063] In another preferred implementation, the pharmaceutical composition of the current invention includes an EZH2 inhibitor. Preferably, the EZH2 inhibitor is selected from the group that consists of: tazemetostat, (S)-1-(sec-butyl)-N-((4,6-dimethyl-2-oxo-1,2-dihidropyridin-3-yl)methyl)-3-methyl-6-(6-(piperazin-1-yl)pihdin-3-yl)-1 H-indol-4-carboxamida, lirametostat, valemetostat, EZM-002, mevrometostat, N-[(6-methyl-2-oxo-4-propyl-1 H-pyridin-3-yl)methyl]-1-propan-2-yl-6-[6-(4-propan-2-ylpiperazin-1- il)pyridin-3-yl]indazol-4-carboxamida, 3-[Ethyl(tetrahydro-2H-pyran-4-yl)amino]-N-[(2,3,5,6,7,8-hexahidro-1-methyl-3-oxo-4- ¡soquinolinyl)methyl]-2-methyl-5-[6-(4-methyl-1-piperazinyl)-3-pyridinyl]-benzamida and OR-S1 , or combinations of los mismos.

[0064] In another preferred embodiment, the pharmaceutical composition of the invention further comprises at least one IL-6 cytokine receptor inhibitor. Preferably, the IL-6 cytokine receptor inhibitor is selected from the group consisting of: tozilizumab, siltuximab, sarilumab, olokizumab, clazakizumab, and levilimab, or combinations thereof.

[0065] In another preferred embodiment, the pharmaceutical composition of the invention further comprises at least one PD-1 receptor inhibitor. Preferably, the PD-1 receptor inhibitor is selected from the group consisting of: nivolumab, pembrolizumab, cemiplimab, dostarlimab, toripalimab, and tislelizumab, or combinations thereof.

[0066] In another preferred embodiment, the pharmaceutical composition of the invention further comprises at least one PD-L1 inhibitor. Preferably, the PD-L1 inhibitor is selected from the group consisting of: atezolizumab, durvalumab, avelumab, and envafolimab, or combinations thereof.

[0067] In another preferred embodiment, the pharmaceutical composition of the invention further comprises at least one pharmaceutically acceptable vehicle or excipient.

[0068] For the purposes of the present invention, “pharmaceutically acceptable excipient” refers to an inert ingredient that stabilizes and promotes the absorption of active ingredients, such as, but not limited to, cosolvents, surfactants, oils, humectants, emollient binders, preservatives, stabilizers, antioxidants, colorants, sweeteners, flavorings, and binders.

[0069] Throughout the description and claims, the terms "comprises," "comprising," and their variants are not limiting in nature and are therefore not intended to exclude other technical features. Specifically, the terms "comprises," "comprising," and their variants throughout the description and claims include the terms "consists of," "consisting of," and their variants.

[0070] As used in this description and in the claims, the singular forms “el”, “la” include references to the plural forms unless the content clearly indicates otherwise.

[0071] BRIEF DESCRIPTION OF THE DRAWINGS

[0072] Fig. 1. Differential protein expression was analyzed between conditioned medium derived from tumor-associated macrophages (M0), triple-negative breast cancer cells (TNBC in the figure), and the co-culture of the two (MO-TNBC in the figure) with an antibody array. The analysis identified four specific and differential cytokines in the conditioned medium derived from the co-culture of tumor-associated macrophages and triple-negative breast cancer cells: CCL3, CCL4, CXCL2, and IL-1. Fig. 2. Quantification by ELISA of the cytokines CCL3, CCL4, CXCL2, and IL-1 in conditioned medium derived from unco-cultured MDA-MB-468 triple-negative breast cancer cells, unco-cultured M0 macrophages, or from the co-culture of both media. In MDA-MB-468 cells, the expression of the KDM6A gene was silenced or 10 pM tazemetostat was added to these cells.

[0073] Fig. 3. Quantification by ELISA of the cytokines CCL3, CCL4, CXCL2 and IL-1 in conditioned medium from unco-cultured MDA-MB-231 triple-negative breast cancer cells, unco-cultured M0 macrophages, or from the co-culture of both media. In MDA-MB-231 cells, the expression of the KDM6A gene was silenced or 10 pM tazemetostat was added to these cells.

[0074] Fig. 4. Differential expression analysis by digital PCR (ddPCR) of the genes involved in the epithelial-mesenchymal transition CDH1, CDH2, TWIST1, and MMP9 in MDA-MB-468 (top row), MDA-MB-231 (middle row), and BT-549 (bottom row) cell lines. Treatment with anakinra or maraviroc reduces the expression of these mesenchymal markers CDH1, CDH2, TWIST1, and MMP9, with the combined treatment of anakinra and maraviroc resulting in the greatest reduction. CM represents conditioned medium.

[0075] Fig. 5. Differential expression analysis by digital PCR (ddPCR) of the genes involved in the epithelial-mesenchymal transition CDH2 and TWIST 1 in the MDA-MB-468 cell line. Anakinra, maraviroc, navahxin, tazemetostat, and combinations thereof were added to MDA-MB-468 cells. CM represents conditioned medium.

[0076] Fig. 6. Cell migration rate of the MDA-MB-468 cell line using a wound closure assay. Anakinra, maraviroc, navahxin, tazemetostat, and combinations thereof were added to MDA-MB-468 cells.

[0077] Fig. 7. Cell migration rate of the MDA-MB-231 cell line using a wound closure assay. MDA-MB-468 cells were treated with anakinra, maraviroc, navahxin, tazemetostat, and combinations thereof.

[0078] Fig. 8. Cell migration rate of BT-549 and BT-20 cell lines using a wound closure assay. The cells were treated with maraviroc, anakinra, and their combination. Fig. 9. A, invasion rate determined in an in vitro tumor cell invasion assay into the surrounding tissue. B, transendothelial migration rate determined in an in vitro transendothelial migration assay after the addition of HlIVEC endothelial cells. The combined treatment with maraviroc and anakinra significantly reduces both the invasion and transendothelial migration processes. CM represents conditioned medium.

[0079] Fig. 10. A, Wound closure assay with MDA-MB-468 cells not previously exposed to macrophage interaction. Cells were treated with fresh medium (control, top row), conditioned medium from the co-culture of M0 macrophages with MDA-MB-468 cells (middle row), or from the co-culture of M0 macrophages with MDA-MB-468 cells, in which the MDA-MB-468 cells were treated with 10 pM tazemetostat during co-culture (bottom row). Images were taken at the indicated times using a microscope at 10X magnification. An enlargement of the indicated area, confirming the changes in cell morphology, is shown at the far left. B, Migration rate determined in the wound closure assay. Tazemetostat is observed to inhibit tumor cell migration. CM represents conditioned medium, and CM TZ represents conditioned medium + tazemetostat.

[0080] Fig. 11. Migration rate determined in a wound closure assay with the MDA-MB-231 cell line. Tazemetostat is observed to inhibit tumor cell migration. CM represents conditioned medium and CM TZ represents conditioned medium + tazemetostat.

[0081] Fig. 12. Migration rate determined in a wound closure assay with the BT-549 cell line. Tazemetostat is observed to inhibit tumor cell migration. CM represents conditioned medium and CM TZ represents conditioned medium + tazemetostat.

[0082] Fig. 13. Migration rate determined in a wound closure assay with the BT-20 cell line. Tazemetostat is observed to inhibit tumor cell migration. CM represents conditioned medium and CM TZ represents conditioned medium + tazemetostat.

[0083] Fig. 14. Gene expression analysis by ddPCR: CD163, VEGFA, PDL2, TWIST1, MMP9 and PD-L1 in lung cancer cell lines (A549 and H1299), prostate cancer (PC3 and LNCAP) and osteosarcoma (U2OS).

[0084] Fig. 15. Quantification by ELISA of the cytokines CCL3, CCL4, CXCL2 and IL-1 p in the conditioned medium from lung cancer cells (A549 and H1299), prostate cancer cells (PC3 and LNCAP) and osteosarcoma cells (U2OS) without co-culture, M0 macrophages without co-culture, or from the co-culture of both media.

[0085] Fig. 16. Migration rate determined in a wound closure assay with lung cancer (A549 and H1299), prostate cancer (PC3), and osteosarcoma (U2OS) cell lines. Anakinra, maraviroc, navahxin, tazemetostat, and combinations thereof were added to the cells. CM represents conditioned medium.

[0086] Fig. 17. Photographs of mice injected with MDA-MB-468 cells to induce metastasis of triple-negative breast cancer. Mouse 1 injected with MDA-MB-468 cells that have not been incubated (stimulated / educated) with macrophages. Mouse 2 injected with MDA-MB-468 cells that have been incubated (stimulated / educated) with macrophages. Mouse 3 injected with MDA-MB-468 cells that have been incubated (stimulated / educated) with macrophages and treated with a combination of Anakinra and Maraviroc (A+M). Mouse 4 injected with MDA-MB-468 cells that have been incubated (stimulated / educated) with macrophages and with Anakinra and Maraviroc (A+M) also present in the incubation medium.

[0087] Fig. 18. Averages obtained in the photographs of the previous mouse groups (7 mice per group) in pixels / second. The total flow measured in pixels / second is proportional to the number of cells in that measurement area and corresponds to pulmonary metastases.

[0088] DESCRIPTION OF REALIZATION MODES

[0089] Example 1. Materials and methods

[0090] Statistical analysis

[0091] The results were generated from three or more independent experiments. Values ​​are presented as mean ± standard error of the mean (SEM). Data with three or more groups were statistically analyzed using one-way analysis of variance and Tukey's multiple comparisons test. For comparisons between two groups, a two-tailed unpaired t-test was used. A p-value < 0.05 was considered statistically significant. In the figures, error bars represent the standard error. In the figures, * represents p < 0.05; ** represents p < 0.01; *** represents p < 0.001; and **** represents p < 0.0001. Example 2. Crosstalk and Cytokine Secretion in Breast Cancer Cells

[0092] A quantitative semantic expression array (Biotin-Label Based Antibody Arrays Human L-493; Human L-507, RayBiotech Life, Inc.) was performed to identify differential proteins between conditioned medium from tumor-associated macrophages (M0), triple-negative breast cancer cells (MDA-MB-468), and the co-culture of the two. Four specific and differential cytokines were identified in the conditioned medium from the co-culture of tumor-associated macrophages and triple-negative breast cancer cells: CCL3, CCL4, CXCL2, and IL-1p (Fig. 1), demonstrating cross-communication between tumor-associated macrophages and triple-negative breast cancer cells.

[0093] These cytokines are fundamental to cancer progression, acting as signals that facilitate communication between tumor cells and macrophages. In this context, macrophages do not attack the tumor, but are instead “reprogrammed” by the tumor environment toward an anti-inflammatory phenotype to support its growth and spread. The cytokines secreted in this interaction strengthen cell-to-cell communication and promote changes in tumor cells that facilitate their dissemination.

[0094] The cytokines CCL3, CCL4, CXCL2, and IL-1 were quantified by ELISA in conditioned medium from unco-cultured MDA-MB-468 triple-negative breast cancer cells, unco-cultured M0 macrophages, or from the co-culture of both. The results are shown in Fig. 2 (MDA-MB-468 cells) and Fig. 3 (MDA-MB-231 cells). In MDA-MB-468 and MDA-MB-231 triple-negative breast cancer cells, the expression of the KDM6A gene was silenced or 10 M tazemetostat was added to these cells. The KDM6A gene encodes an epigenetically regulated enzyme, the histone H3 lysine 27 demethylase.

[0095] Figures 2 and 3 show that conditioned medium from unco-cultured cell lines (MDA-MB-468 and MDA-MB-231) and unco-cultured M0 macrophages secrete lower levels of all cytokines. In contrast, conditioned medium from the co-culture of M0 macrophages and triple-negative breast cancer cells secretes higher levels of all cytokines, which are reduced when the co-culture has been previously carried out in the presence of tazemetostat. Example 3. Expression of genes involved in the epithelial-mesenchymal transition in breast cancer cells

[0096] In the epithelial-mesenchymal transition, epithelial cells acquire mesenchymal characteristics, including a greater capacity for migration and invasion, and allow tumor cells to detach from the primary tumor and resist apoptosis.

[0097] A differential expression analysis of genes involved in the epithelial-mesenchymal transition was performed using Droplet Digital PCR (ddPCR). Following treatment with conditioned medium from the co-culture of triple-negative breast cancer cells (cell lines MDA-MB-468, MDA-MB-231, and BT-549) with M0 macrophages, an increase in mesenchymal markers was observed, indicating that the secreted cytokines CCL3, CCL4, CXCL2, and IL-1β induce the epithelial-mesenchymal transition, thereby increasing the metastatic capacity of the tumor cells.

[0098] Treatment with 2 pM anakinra or 20 nM maraviroc reduces the expression of the mesenchymal markers CDH1, CDH2, TWIST1, and MMP9 in the MDA-MB-468, MDA-MB-231, and BT-549 cell lines (Fig. 4). The combined treatment of anakinra and maraviroc reduces the expression of the mesenchymal markers analyzed in this example to the greatest extent.

[0099] Treatment with anakinra 2 pM, maraviroc 20 nM, navarixin 20 nM, tazemetostat 10 pM and combinations thereof reduces the expression of the mesenchymal markers CDH2 and TWIST1 in the MDA-MB-468 cell line (Fig. 5).

[0100] Example 4. Reduction of cell migration of breast cancer cells

[0101] Cell migration is the first step in metastasis, allowing cancer cells to move from the primary tumor to other tissues. Cytokines act as chemoattractants, creating chemical gradients that direct the movement of tumor cells. This process is crucial for cancer cells to invade adjacent tissues and eventually enter the bloodstream or lymphatic system, facilitating their spread.

[0102] The cell migration rate of triple-negative breast cancer cell lines was determined using a wound-closure assay. The wound-closure assay is a method for studying directional cell migration in vitro. This method simulates cell migration during wound closure in vivo. In this assay, a “wound” is created in a cell monolayer, images are captured at the beginning and at regular intervals during cell migration to close the wound, and the images are compared to quantify the cell migration rate. The wound-closure assay was performed for 12 hours, using a 10X objective and taking images every 3 hours with a Leica DM18 timelapse microscope.

[0103] Treatment with anakinra 2 pM, maraviroc 20 nM, navarixin 20 nM, tazemetostat 10 pM and combinations thereof reduces the migration capacity of the MDA-MB-468 (Fig. 6) and MDA-MB-231 (Fig. 7) cell lines.

[0104] In the MDA-MB-468 cell line, treatment with maraviroc alone reduces cell migration capacity by 50%, anakinra by 75%, and navarixin by 25%; the combination of all three drugs reduces it by approximately 80% (Fig. 6). The combination of the three drugs is more effective than individual treatment in inhibiting the cell migration process.

[0105] Treatment with either maraviroc 20 nM or anakinra 2 pM individually reduces cell migration capacity in the BT-549 (Fig. 8, left) and BT-20 (Fig. 8, right) cell lines. The combination of anakinra and maraviroc is more effective than treatment with either drug individually in inhibiting cell migration (Fig. 8).

[0106] Example 5. Reduction of invasion and transendothelial migration of breast cancer cells

[0107] Invasion is a critical step in metastasis, where cancer cells penetrate and degrade the extracellular matrix. Cytokines play a fundamental role by activating proteolytic enzymes such as matrix metalloproteinases, which degrade extracellular matrix components, facilitating the invasion of tumor cells into new tissues. Transendothelial migration, or transvasation, is the process by which tumor cells cross the walls of blood or lymphatic vessels to enter the circulation. This step is essential for cancer cells to travel through the bloodstream or lymphatic system and colonize distant organs. An in vitro tumor cell invasion assay was performed in the surrounding tissue for 24 hours. An in vitro transendothelial migration or transvasation assay was also performed for 24 hours, in which HIIVEC endothelial cells were added to mimic the in vivo process.Cytokines facilitate this process by promoting vascular permeability and allowing tumor cells to interact with the vascular endothelium. Once in circulation, these cells can migrate to other organs, extravasate, and form secondary metastases.

[0108] Both assays (cell invasion assay and transendothelial migration assay) were performed with 8 pM pore size insert systems, using the conditioned medium as a chemoattractant in the well, and in both assays Geltrex™ membrane matrices (Gibco) (0.12 mg / ml) were used to mimic the extracellular matrix in the 8 pM pore size insert.

[0109] In the cell invasion and transendothelial migration assays described in the preceding paragraphs, the combined treatment of maraviroc 20 nM and anakinra 2 pM significantly reduced both the invasion process (Fig. 9A) and the transendothelial migration process (Fig. 9B).

[0110] Example 6. Reduction of cell migration of breast cancer cells with tazemetostat

[0111] The cell migration rate of triple-negative breast cancer cells MDA-MB-468 (Fig. 10), MDA-MB-231 (Fig. 11), BT-549 (Fig. 12), and BT-20 (Fig. 13) was determined using a wound closure assay as described in Example 4, in this case, with the addition of 10 pM tazemetostat. The conditioned medium from the co-culture of triple-negative breast cancer cells with M0 macrophages activates tumor cell migration, which is inhibited after treatment with tazemetostat for three days of co-culture.

[0112] Example 7. Expression of genes involved in invasion and immunosuppression in lung, prostate and osteosarcoma cancer cells

[0113] Gene expression analysis of those involved in invasion and immunosuppression was performed using ddPCR. Following treatment with conditioned medium from the co-culture of lung cancer cells (A549 and H1299), prostate cancer cells (PC3 and LNCAP), and osteosarcoma cells (U2OS) with M0 macrophages, an increase in the polarization of macrophages from M0 to pro-tumoral M2 was observed, as evidenced by an increase in the expression of the markers CD-163, VEGFA, and PD-L2, and the M2 subtype is characterized by an increase in these markers. An increase in the expression of invasion and immunosuppression markers was also observed, specifically an increase in PD-L1 (an immunosuppression marker), VEGFA (an angiogenesis marker), MMP9 (an invasion marker), and TWIST1 (an epithelial-mesenchymal transition marker) (Fig. 14).

[0114] Example 8. Cytokine secretion in lung, prostate, and osteosarcoma cancer cells

[0115] The cytokines CCL3, CCL4, CXCL2 and I L-1 p were quantified by ELISA in the conditioned medium from lung cancer cells (A549 and H1299), prostate cancer cells (PC3 and LNCAP) and osteosarcoma cells (U2OS) without co-culture, M0 macrophages without co-culture, or from the co-culture of both and the results are shown in Fig. 15.

[0116] Figure 15 shows that conditioned medium from all non-co-cultured cell lines and non-co-cultured M0 macrophages secreted lower levels of all cytokines. In contrast, conditioned medium from the co-culture of M0 macrophages and cells from lung cancer, prostate cancer, and osteosarcoma cell lines secreted higher levels of all cytokines.

[0117] Example 9. Reduction of cell migration of lung, prostate, and osteosarcoma cancer cells

[0118] The cell migration rate of lung cancer cell lines (A549 and H1299), prostate cancer (PC3 and LNCAP) and osteosarcoma (U2OS) has been determined by a wound closure assay as described in Example 4.

[0119] Treatment with anakinra 2 pM, maraviroc 20 nM, navarixin 20 nM, tazemetostat 10 pM and combinations thereof reduces the migration capacity of the tested lung cancer, prostate cancer and osteosarcoma cells (Fig. 16).

[0120] Example 10. Representative in vivo images of bioluminescence and drug treatment efficacy in TNBC cells conditioned / stimulated / educated by macrophages.

[0121] Representative images (Fig. 17) were acquired using a preclinical multimodal imaging system, the Lumina X5 Imaging System (PerkinElmer), on mice (7 per group). Mice in group 1 were intravenously injected with MDA-MB-468 cells previously cultured for three days in conditioned medium (1:3 dilution with fresh medium), without macrophages present in the culture medium (Cells-468). Mice in group 2 were intravenously injected with macrophage-educated MDA-MB-468 cells for three days, but without Anakinra or Maraviroc (A+M) in the culture medium. Mice in group 3 were intravenously injected with “macrophage-educated” MDA-MB-468 cells for three days and then treated daily for five consecutive days with intraperitoneal injections of 20 mg / kg of Maraviroc plus 25 mg / kg of Anakinra (A+M).Mice in group 4 were intravenously injected with macrophage-educated MDA-MB-468-Luc cells for three days in the presence of (A+M) in the culture medium. Bioluminescence was quantified at the indicated time points after intraperitoneal injection of 150 mg / kg of D-luciferin and image acquisition 20 minutes later (1-minute exposure), with pixels per second measured on the y-axis. Figure 18 shows the average ± SEM measurements obtained for each mouse group. All animal experiments were conducted in accordance with European and Spanish regulations and were approved by the Institutional Committee for Animal Experimentation of the Valencian Government.

[0122] The injection of MDA-MB-468 cells produces metastasis (darker patches in mice, in shades of gray). This metastasis increases when the triple-negative breast cancer cell line (MDA-MB-468 cells) is incubated / stimulated / conditioned / "educated" in the presence of macrophages (Mouse Group 2). However, treatment of the mice, once they have received the injection of cancer cells, with the combination of Anakinra and Maraviroc (A+M) almost completely prevents the appearance of metastasis (Mouse Group 3).Moreover, the prior incubation of these triple-negative breast cancer cells (MDA-MB-468 cells), even when stimulated or "educated" with macrophages, which makes them even more virulent in generating metastases, in the presence of the combination of Anakinra and Maraviroc (A+M), completely prevents the appearance of metastases; that is, the combination (A+M) present in the incubation medium of the MDA-MB-468 cells that are being "educated" with the macrophages also present in this culture medium, prevents the cancer cells from being "educated" and thus losing their metastatic capacity (Mouse Group 4).

Claims

CLAIMS 1. A combination of at least two cytokine receptor inhibitors or a pharmaceutical composition comprising them, for use in the prevention or treatment of metastasis, wherein the cytokines consist of IL-1p, CCR5 or CXCR2.

2. The combination or composition for use according to claim 1, characterized in that at least one of the two inhibitors present in the combination is an inhibitor of the cytokine I L-1 p receptor.

3. The combination or composition for use according to claim 2, characterized in that the IL-1p cytokine receptor inhibitor is selected from the group consisting of: anakinra, canakinumab, rilonacept, belsomra, gevokizumab, lutikizumab, GSK 1070806, dapansutrile and zerumbona, or combinations thereof.

4. The combination or composition for use according to claim 2 or 3, characterized in that the IL-1p cytokine receptor inhibitor is anakinra.

5. The combination or composition for use according to claim 1, characterized in that at least one of the two inhibitors present in the combination is an inhibitor of the CCR5 cytokine receptor.

6. The combination or composition for use according to claim 5, characterized in that the CCR5 cytokine receptor inhibitor is selected from the group consisting of: maraviroc, Met-CCL3, Met-CCL3 / CCL4, plerixafor, cucurbitacin I, leronlimab, naltrexone, dimethyl-[[4-[[3-(4-methylphenyl)-8,9-dihydro-7H-benzo[7]annulene-6-carbonyl]amino]phenyl]methyl]-(oxan-4-yl)azanium and 1 [2- [4- (trif I u orom eti l)fe ni I ]et¡ l]es pi ro[1 H-3, 1 -benzoxazine-4,4'-piperidine]-2-one, or combinations thereof.

7. The combination or composition for use according to claim 5 or 6, characterized in that the CCR5 cytokine receptor inhibitor is maraviroc.

8. The combination or composition for use according to claim 1, characterized in that at least one of the two inhibitors present in the combination is an inhibitor of the CXCR2 cytokine receptor.

9. The combination or composition for use according to claim 8, characterized in that the CXCR2 cytokine receptor inhibitor is selected from the group consisting of: navarixin, danirixin, elubrixin and reparixin, or combinations thereof.

10. The combination or composition for use according to claim 9, characterized in that the CXCR2 cytokine receptor inhibitor is navarixin.

11. The combination or composition for use according to any of claims 1 to 10, characterized in that said combination or composition comprises at least one inhibitor of the cytokine I L-1 p receptor, at least one inhibitor of the cytokine receptor CCR5 and at least one inhibitor of the cytokine receptor CXCR2.

12. The combination or composition for use according to claim 11, characterized in that the cytokine I L-1 receptor inhibitor is anakinra, the CCR5 cytokine receptor inhibitor is maraviroc, and the CXCR2 cytokine receptor inhibitor is navarixin.

13. The combination or composition for use according to any of claims 1 to 12, characterized in that said combination or composition further comprises at least one EZH2 inhibitor.

14. The combination or composition for use according to claim 13, characterized in that the EZH2 inhibitor is selected from the group consisting of: tazemetostat, (S)-1-(sec-butyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-methyl-6-(6-(piperazin- 1-yl)pyridin-3-yl)-1 H-indole-4-carboxamide, lirametostat, valemetostat, EZM-002, mevrometostat, N-[(6-methyl-2-oxo-4-propyl-1 H-pyridin-3-yl)methyl]-1-propan-2-yl-6-[6-(4-propan-2-ylpiperazin- 1-yl)pyridin-3-yl]indazol-4-carboxamide, and OR-S1, or combinations thereof.

15. The combination or composition for use according to claim 14, characterized in that the EZH2 inhibitor is tazemetostat.

16. The combination or composition for use according to claims 1 to 7, characterized in that the at least two cytokine inhibitors are anakinra and maraviroc.

17. The combination or composition for use according to claim 16, in the prevention or treatment of metastasis originating from triple-negative breast cancer.

18. The combination or composition for use according to claims 1 to 4 and 8 to 10, characterized in that the at least two cytokine inhibitors are anakinra and navarixin.

19. The combination or composition for use according to claim 18, in the prevention or treatment of metastasis originating from triple-negative breast cancer.

20. The combination or composition for use according to claims 5 to 10, characterized in that the at least two cytokine inhibitors are maraviroc and navarixin.

21. The combination or composition for use according to claim 20 in the prevention or treatment of metastasis originating from triple-negative breast cancer.

22. The combination or composition for use according to any of claims 1 to 21, characterized in that said combination or composition further comprises at least one IL-6 cytokine receptor inhibitor.

23. The combination or composition for use according to claim 22, characterized in that the IL-6 cytokine receptor inhibitor is selected from the group consisting of: tozilizumab, siltuximab, sarilumab, olokizumab, clazakizumab and levilimab, or combinations thereof.

24. The combination or composition for use according to any of claims 1 to 23, characterized in that said combination or composition further comprises at least one PD-1 receptor inhibitor.

25. The combination or composition for use according to claim 24, characterized in that the PD-1 receptor inhibitor is selected from the group consisting of: nivolumab, pembrolizumab, cemiplimab, dostarlimab, toripalimab and tislelizumab, or combinations thereof.

26. The combination or composition for use according to any of claims 1 to 25, characterized in that said combination or composition further comprises at least one PD-L1 inhibitor.

27. The combination or composition for use according to claim 26, characterized in that the PD-L1 inhibitor is selected from the group consisting of: atezolizumab, durvalumab, avelumab and envafolimab, or combinations thereof.

28. The combination or composition for use according to any of claims 1 to 27, characterized in that the metastasis originated from a tumor selected from the group consisting of: prostate cancer, lung cancer, and osteosarcoma.

29. The combination or composition for use according to any of claims 1 to 27, characterized in that the metastasis originated in triple-negative breast cancer.

30. The combination or composition for use according to any of claims 1 to 29, in a subject who has undergone tumor resection or biopsy.

31. The combination or composition for use according to any of claims 1 to 30, in a cancer or tumor that does not respond to chemotherapy.

32. The composition for use according to any of claims 1 to 31, characterized in that said composition further comprises at least one pharmaceutically acceptable vehicle or excipient.

33. A pharmaceutical composition comprising at least two cytokine receptor inhibitors, wherein the cytokines consist of IL-1p, CCR5 or CXCR2.

34. The pharmaceutical composition according to claim 33, characterized in that at least one of the two inhibitors present in the composition is an inhibitor of the cytokine I L-1 p receptor.

35. The pharmaceutical composition according to claim 34, characterized in that the IL-1 cytokine receptor inhibitor is selected from the group consisting of: anakinra, canakinumab, rilonacept, belsomra, gevokizumab, lutikizumab, GSK 1070806, dapansutrilo and zerumbona, or combinations thereof.

36. The pharmaceutical composition according to claim 34 or 35, characterized in that the IL-1p cytokine receptor inhibitor is anakinra.

37. The pharmaceutical composition according to claim 33, characterized in that at least one of the two inhibitors present in the combination is a CCR5 cytokine receptor inhibitor.

38. The pharmaceutical composition according to claim 37, characterized in that the CCR5 cytokine receptor inhibitor is selected from the group consisting of: maraviroc, Met-CCL3, Met-CCL3 / CCL4, plerixafor, cucurbitacin I, leronlimab, naltrexone, dimethyl-[[4-[[3-(4-methylphenyl)-8,9-dihydro-7H-benzo[7]annulene-6-carbonyl]amino]phenyl]methyl]-(oxan-4-yl)azanium and 1 [2- [4- (trif I u orom eti l)fe ni I ]et¡ l]es pi ro[1 H-3, 1 -benzoxazine-4,4'-piperidine]-2-one, or combinations thereof.

39. The pharmaceutical composition according to claim 37 or 38 characterized in that the CCR5 cytokine inhibitor is maraviroc.

40. The pharmaceutical composition according to claim 33, characterized in that at least one of the two inhibitors present in the combination is an inhibitor of the CXCR2 cytokine receptor.

41. The pharmaceutical composition according to claim 40, characterized in that the CXCR2 cytokine receptor inhibitor is selected from the group consisting of: navarixin, danirixin, elubrixin and reparixin, or combinations thereof.

42. The pharmaceutical composition according to claim 40 or 41, characterized in that the CXCR2 cytokine receptor inhibitor is navarixin.

43. The pharmaceutical composition according to any of claims 33 to 42, characterized in that said pharmaceutical composition comprises at least one inhibitor of the cytokine receptor I L-1 p, at least one inhibitor of the cytokine receptor CCR5 and at least one inhibitor of the cytokine receptor CXCR2.

44. The pharmaceutical composition according to claim 43, characterized in that the IL-1 cytokine receptor inhibitor is anakinra, the CCR5 cytokine receptor inhibitor is maraviroc, and the CXCR2 cytokine receptor inhibitor is navarixin.

45. The pharmaceutical composition according to any of claims 33 to 44, characterized in that said pharmaceutical composition further comprises at least one EZH2 inhibitor compound.

46. ​​The pharmaceutical composition according to claim 45, characterized in that the EZH2 inhibitor is selected from the group consisting of: tazemetostat, (S)-1-(sec-butyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-methyl-6-(6-(piperazin- 1-yl)pyridin-3-yl)-1 H-indole-4-carboxamide, lirametostat, valemetostat, EZM-002, mevrometostat, N-[(6-methyl-2-oxo-4-propyl-1 H-pyridin-3-yl)methyl]-1-propan-2-yl-6-[6-(4-propan-2-ylpiperazin- 1-yl)pyridin-3-yl]indazol-4-carboxamide, and OR-S1, or combinations thereof.

47. The pharmaceutical composition according to any of claims 33 to 46 wherein the at least two cytokine inhibitors are anakinra and maraviroc.

48. The pharmaceutical composition according to any of claims 33 to 46 wherein the at least two cytokine inhibitors are anakinra and navaraxin.

49. The pharmaceutical composition according to any of claims 33 to 46 wherein the at least two cytokine inhibitors are maraviroc and navaraxin.

50. The pharmaceutical composition according to any of claims 33 to 49 comprising anakinra, maraviroc and navaraxin.

51. The pharmaceutical composition according to claim 50 further comprising tazemetostat.

52. The pharmaceutical composition according to any of claims 33 to 51, characterized in that said pharmaceutical composition further comprises at least one IL-6 cytokine receptor inhibitor.

53. The pharmaceutical composition according to claim 52, characterized in that the IL-6 cytokine receptor inhibitor is selected from the group consisting of: tozilizumab, siltuximab, sarilumab, olokizumab, clazakizumab and levilimab, or combinations thereof.

54. The pharmaceutical composition according to any of claims 33 to 53, characterized in that said pharmaceutical composition further comprises at least one PD-1 receptor inhibitor.

55. The pharmaceutical composition according to claim 54, characterized in that the PD-1 receptor inhibitor is selected from the group consisting of: nivolumab, pembrolizumab, cemiplimab, dostarlimab, toripalimab and tislelizumab, or combinations thereof.

56. The pharmaceutical composition according to any of claims 33 to 55, characterized in that said pharmaceutical composition further comprises at least one PD-L1 inhibitor.

57. The pharmaceutical composition according to claim 56, characterized in that the PD-L1 inhibitor is selected from the group consisting of: atezolizumab, durvalumab, avelumab and envafolimab, or combinations thereof.

58. The pharmaceutical composition according to any of claims 33 to 57, further comprising at least one pharmaceutically acceptable vehicle or excipient.

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