Methods for the treatment of chemotherapy-induced cardiotoxicity in a subject in need thereof

Abstract

The evidences provided by the present patent application demonstrate the therapeutic effect of succinate dehydrogenase inhibition to counteract the cardiotoxicity of chemotherapies used in cancer, such as acute myeloid leukemia and breast cancer. This combination also remarkably exhibits a chemosensitizing capacity of cancer cells. The use of Malonate in patients affected by these pathologies would therefore protect the heart from the toxic side effects of chemotherapy and eliminate the most resistant cells for an ever better risk-benefit ratio for these patients. Accordingly, the present invention relates to a method for the treatment of chemotherapy-induced cardiotoxicity in a subject in need thereof comprising administering to said subject a therapeutically effective amount of a succinate dehydrogenase inhibitor.

Description

[0001] METHODS FOR THE TREATMENT OF CHEMOTHERAPY-INDUCED CARDIOTOXICITY IN A SUBJECT IN NEED THEREOF

[0002] FIELD OF THE INVENTION:

[0003] The present invention is in the field of medicine, in particular cardio-oncology.

[0004] BACKGROUND OF THE INVENTION:

[0005] A large number of treatments used in oncology are cardiotoxic and can lead to the emergence of cardiovascular disease (CVD). These can force the medical profession to reduce or even stop treatment for patients, and are the cause of many short- and medium-term deaths1. The remarkable improvement in the therapeutic management of certain cancers has led to a significant increase in these patients' chances of survival. For some cancers, this has led to disease-related deaths being surpassed by deaths caused by treatment-related cardiotoxicity2.

[0006] Although anthracyclines increase long-term survival after cancer, a dose-dependent relationship between anthracy cline dose and cardiovascular complications has been reported1. Unfortunately, even targeted therapies (kinase inhibitors) cause up to 44% of heart disease via mechanisms similar to anthracyclines3,4. In contrast to tumor cells, cardiomyocytes are terminally differentiated cell type with limited turnover explaining the long-term effects of such cytotoxic medications. It is therefore essential to specifically target the pathological processes at the root of these major complications.

[0007] The mechanisms underlying anthracyclines cardiotoxicity (such as doxorubicin) are well known. Including senescence and apoptosis of various cell types via increased production of Reactive Oxygen Species (ROS) due to massive metabolic reprogramming and mitochondrial dysfunction, leading to functional defects in cardiac and vascular tissue.

[0008] Recent work suggests that the use of oxidative phosphorylation metabolism inhibitors (or OXPHOSi) may protect the heart from the toxic effects of doxorubicin5. Interestingly, complex II of the electron transfer chain (ETC) has never been studied for cardioprotective strategies, even though this complex is preferentially fed by fatty acid metabolism, the heart's main energy source. Yet succinate (substrate of complex II or succinate dehydrogenase; SDH) is strongly linked to the emergence of CVD6. Malonate, an SDH inhibitor, reduces the effects of ischemia / reperfusion in various organs (including the heart) by decreasing ROS production6,7. By inhibiting SDH, Malonate prevents ETC reversion and thus electron influx to complex I, synonymous with ROS production.

[0009] Combinations of chemotherapy and OXPHOSi have opened up a new avenue of research for the treatment of particularly aggressive cancers. These OXPHOSi potentiate the effect of these therapies by sensitizing cancer cells, which are usually dependent on their mitochondrial metabolism to resist treatment. Thus, several OXPHOSi are currently being used in preclinical in vivo models8,9and in clinical trials in various cancers. Finally, ETC reversion has been shown to be a key mechanism of tumorigenesis10and SDH inhibition is effective in eliminating cancer cells11.

[0010] SUMMARY OF THE INVENTION:

[0011] The invention is defined by the claims. In particular, the present invention relates to a method for the treatment of chemotherapy-induced cardiotoxicity in a subject in need thereof comprising administering to said subject a therapeutically effective amount of a succinate dehydrogenase inhibitor.

[0012] DETAILED DESCRIPTION OF THE INVENTION:

[0013] The first object of the present invention relates to a method for the treatment of chemotherapy -induced cardiotoxicity in a subject in need thereof comprising administering to said subject a therapeutically effective amount of a succinate dehydrogenase inhibitor.

[0014] As used herein, the term “subject” or “patient” denotes a mammal, preferably human. Typically, a subject according to the invention refers to any subject (preferably human) afflicted with cancer. Other subjects may include cattle, dogs, cats, guinea pigs, rabbits, rats, mice, horses, and the like. In some embodiments, the subject is a human. In some embodiments, the succinate dehydrogenase inhibitor of the present invention is administered to a subject having one or more signs or symptoms of cardiotoxicity due to chemotherapy (e.g. any functional or structural heart damage arising from cancer treatment). As example, such symptoms includes shortness of breath, chest pain, heart palpitations, fluid retention in the legs, distention of the stomach, dizziness during or after chemotherapy. As used herein, the term "treatment" or "treat" refer to both prophylactic or preventive treatment as well as curative or disease modifying treatment, including treatment of patient at risk of contracting the disease or suspected to have contracted the disease as well as patients who are ill or have been diagnosed as suffering from a disease or medical condition, and includes suppression of clinical relapse. The treatment may be administered to a subject having a medical disorder or who ultimately may acquire the disorder, in order to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of a disorder or recurring disorder, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment. By "therapeutic regimen" is meant the pattern of treatment of an illness, e.g., the pattern of dosing used during therapy. A therapeutic regimen may include an induction regimen and a maintenance regimen. The phrase "induction regimen" or "induction period" refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the initial treatment of a disease. The general goal of an induction regimen is to provide a high level of drug to a patient during the initial period of a treatment regimen. An induction regimen may employ (in part or in whole) a "loading regimen", which may include administering a greater dose of the drug than a physician would employ during a maintenance regimen, administering a drug more frequently than a physician would administer the drug during a maintenance regimen, or both. The phrase "maintenance regimen" or "maintenance period" refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the maintenance of a patient during treatment of an illness, e.g., to keep the patient in remission for long periods of time (months or years). A maintenance regimen may employ continuous therapy (e.g., administering a drug at a regular interval, e.g., weekly, monthly, yearly, etc.) or intermittent therapy (e.g., interrupted treatment, intermittent treatment, treatment at relapse, or treatment upon achievement of a particular predetermined criteria [e.g., disease manifestation, etc.]).

[0015] By a "therapeutically effective amount" is meant a sufficient amount of the active ingredient for treating or reducing the symptoms at reasonable benefit / risk ratio applicable to any medical treatment. It will be understood that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject will depend upon a variety of factors including the disorder being treated and the severity of the disorder; activity of the specific compound employed; the specific composition employed, the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination with the active ingredients; and like factors well known in the medical arts. For example, it is well within the skill of the art to start doses of the compound at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved. However, the daily dosage of the products may be varied over a wide range from 0.01 to 1,000 mg per adult per day. Typically, the compositions contain 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 250 and 500 mg of the active ingredient for the symptomatic adjustment of the dosage to the subject to be treated. A medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, typically from 1 mg to about 100 mg of the active ingredient. An effective amount of the drug is ordinarily supplied at a dosage level from 0.0002 mg / kg to about 20 mg / kg of body weight per day, especially from about 0.001 mg / kg to 7 mg / kg of body weight per day.

[0016] As used herein, the term “chemotherapy-induced cardiotoxicity” has its general meaning in the art and refers to any functional or structural heart damage arising from a chemotherapy. A cardioprotective agent is therefore needed. As used herein the term "cardioprotective" means protecting against or reducing damage to the myocardium due to chemotherapy, after, during or prior to chemotherapy. The method of the present invention is particularly suitable for preventing the left ventricle ejection fraction (LVEF) reduction, preventing fibrosis and / or preventing cardiomyocyte cell death in a subject after, during or prior chemotherapy.

[0017] In another aspect, the present invention also relates to a method for the treatment of a chemoresistant cancer in a subject in need thereof comprising administering to said subject a therapeutically effective amount of a succinate dehydrogenase inhibitor and at least one chemotherapeutic agent (i.e. a chemotherapy). In some embodiments, the chemoresistant cancer is a breast cancer or an acute myeloid leukemia. In some embodiments, the chemoresistant cancer is a breast cancer and the at least one chemotherapeutic agent is doxorubicine and cyclophosphamide. In some embodiments, the chemoresistant cancer is an acute myeloid leukemia and the at least one chemotherapeutic agent is doxorubicine and cytarabine. As used herein, the term “chemotherapy” has its general meaning in the art and refers to a treatment that consists in administering to the patient at least one chemotherapeutic agent (i.e. 1, 2, 3, 4, 5 or more chemotherapeutic agent). Chemotherapeutic agents include, but are not limited to Doxorubicin, Cytarabine, Cyclophosphamide, Azacitidine, Carboplatin, Paclitaxel, Methotrexate, Pemetrexed, 5 -Fluorouracil, Capecitabine, Gemcitabine, Venetoclax, Trifluridine, 6-Mercaptopurine, Azathioprine, Fludarabine, Cladribine, Clofarabine, Floxuridine, Dacarbazine, Pentostatin, Pralatrexate, Thioguanine, Nelarabine, Hydroxyurea, Ifosfamide, Chlorambucil, Melphalan, Temozolomide, Thiotepa, Trabectedin, Carmustine, Lomustine, Decitabine, Streptozocin, Mechlorethamine, Altretamine, Bendamustine, Busulfan, Procarbazine, Prednisone, Methylprednisolone, Mitoxantrone, Teniposide, Dexamethasone, Cisplatin, Carboplatin, Oxaliplatin, Irinotecan, Topotecan, Etoposide, Vincristine, Vinblastine, Vinorelbine, Docetaxel, Nab-Paclitaxel, Paclitaxel, Eribulin, Ixabepilone, Epothilone, Bleomycin, Dactinomycin, Mytomycin-C, Actinomycin D, Daunorubicin, Epirubicin, Idarubicin, Valrubicin, Mitomycin, Imatinib, Nilotinib, Erlotinib, Gefitinib, Afatinib, Transretinoic acid, arsenic trioxide, Mitotane, Osimertinib, Omacetaxine, Cabozantinib, Pazopanib, Sunitinib, Sorafenib, Tivozanib, Axitinib, Lenvatinib, Regorafenib, Vandetanib, Alectinib, Crizotinib, Dabrafenib, Encorafenib, Vemurafenib, Trametinib, Trastuzumab, Romidepsin, Vorinostat, Pegasparhase, Cabazitaxel, Ibrutinib, Ruxolitinib, L-Asparaginase, Bortezomib, Carfilzomib, Ixazomib or Olaparib and any pharmaceutically acceptable salts, acids or derivatives of any of the above, or any combination of any of the above. In some embodiments, the at least one chemotherapeutic agent is selected from the group comprising alkylating agents, antimetabolites (e.g. pyrimidine analogs or purine analogs), topoisomerase inhibitors, antitumor antibiotics, mitotic inhibitors, nitrosoureas, corticosteroids or protein kinase inhibitors or any combination thereof.

[0018] In some embodiments, the at least one chemotherapeutic agent is an anthracycline. In some embodiments, the at least one chemotherapeutic agent is doxorubicin. In some embodiments, the at least one chemotherapeutic agent is an anthracycline and an antimetabolite or an alkylating agent. In some embodiments, the at least one chemotherapeutic agent is doxorubicin and cytarabine. In some embodiments, the at least one chemotherapeutic agent is doxorubicin and cyclophosphamide.

[0019] In some embodiments, the at least one chemotherapeutic agent is a BTK (Bruton’s tyrosine kinase) inhibitor. In some embodiments, the at least one chemotherapeutic agent is Ibrutinib. In some embodiments, the at least one chemotherapeutic agent is a MEK inhibitor. In some embodiments, the at least one chemotherapeutic agent is Trametinib. In some embodiments, the at least one chemotherapeutic agent is a BTK inhibitor and / or a MEK inhibitor. In some embodiments, the at least one chemotherapeutic agent Ibrutinib and / or Trametinib.

[0020] In some embodiments, the at least one chemotherapeutic agent is a Bcl-2 inhibitor. In some embodiments, the at least one chemotherapeutic agent is Venetoclax.

[0021] As used herein, the term “succinate dehydrogenase” or “SDH” has its general meaning in the art and refers to an enzyme complex involved in both the tricarboxylic acid cycle and electron transport chain. SDH comprises four structurally different subunits : SdhA (NCBI Gene : 6389; Ensembl: ENSG00000073578), SdhB (NCBI Gene : 6390; ENSG00000117118), SdhC (NCBI Gene : 6391; Ensembl : ENSG00000143252), SdhD (NCBI Gene : 6392; Ensembl : ENSG00000204370).

[0022] As used herein, the term “succinate dehydrogenase inhibitor” denotes a molecule that partially or totally inhibits the biological activity (e.g. dehydrogenase activity) or expression of SDH (e.g. SDH subunits). Such inhibitors includes small organic molecules, antisense oligonucleotides, ribozymes, siRNAs, endonucleases, antibodies (e.g. single domain antibody) directed against any SDH subunits, aptamers or polypeptides. In some embodiments, the SDH inhibitor interacts directly with SDH.

[0023] In some embodiments, the SDH inhibitor according to the invention may be a low molecular weight compound, e. g. a small organic molecule (natural or not). The term "small organic molecule" refers to a molecule (natural or not) of a size comparable to those organic molecules generally used in pharmaceuticals. The term excludes biological macromolecules (e. g., proteins, nucleic acids, etc.). Preferred small organic molecules range in size up to about 10000 Da, more preferably up to 5000 Da, more preferably up to 2000 Da and most preferably up to about 1000 Da. Succinate dehydrogenase inhibitors are well known in the art and includes ME1111 (Meiki Seika Pharma, Cas No 1391758-52-3), malonate (Cas No 156-80-9), disodium malonate (Cas No 141-95-7), dimethyl malonate (Cas No 108-59-8), malate (Cas 149-61-1), 3- nitropropionic acid (Cas No 504-88-1) or oxaloacetate (Cas No 149-63-3), Atpenin A5 (Cas No 119509-24-9) or any derivatives or pharmaceutically acceptable salts thereof. The term “derivatives” has its general meaning in the art and refers to a compound that is formed from a similar compound or a compound that can arise from another atom if one atom is replaced with another atom or group of atoms.

[0024] The term “pharmaceutically acceptable salt” has its general meaning in the art and refers to salts which are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like.

[0025] In some embodiments, the present invention also relates to a method for the preventive treatment of chemotherapy -induced cardiotoxicity in a subject in need thereof comprising administering to said subject a therapeutically effective amount of a succinate dehydrogenase inhibitor. In some embodiments, the present invention also relates to a method for the preventive treatment of chemotherapy -induced cardiotoxicity in a subject in need thereof comprising administering to said subject a therapeutically effective amount of a succinate dehydrogenase inhibitor in combination with at least one chemotherapeutic agent. In some embodiments, the succinate dehydrogenase inhibitor is administrated simultaneously, separately or sequentially with at least one chemotherapeutic agent.

[0026] In some embodiments, the present invention relates to i) a succinate dehydrogenase inhibitor, and ii) at least one chemotherapeutic agent, as a combined preparation for simultaneous, separate or sequential use in the treatment of chemotherapy-induced cardiotoxicity in a subject suffering from cancer.

[0027] As used herein, the term “simultaneous use” denotes the use of a succinate dehydrogenase inhibitor and at least one chemotherapeutic agent occurring at the same time. As used herein, the term “separate use” denotes the use of a succinate dehydrogenase inhibitor and at least one chemotherapeutic agent not occurring at the same time. As used herein, the term “sequential use” denotes the use of a succinate dehydrogenase inhibitor and at least one chemotherapeutic agent occurring by following an order.

[0028] As used herein, the term "cancer" has its general meaning in the art and includes, but is not limited to, solid tumors and blood borne tumors. The term cancer includes diseases of the skin, tissues, organs, bone, cartilage, blood and vessels. The term "cancer" further encompasses both primary and metastatic cancers. Examples of cancers include, but are not limited to, cancer cells from the bladder, blood, bone, bone marrow, brain, breast, colon, esophagus, gastrointestine, gum, head, kidney, liver, lung, nasopharynx, neck, ovary, prostate, skin, stomach, testis, tongue, or uterus. In addition, the cancer may specifically be of the following histological type, though it is not limited to these: neoplasm, malignant; carcinoma; carcinoma, undifferentiated; giant and spindle cell carcinoma; small cell carcinoma; papillary carcinoma; squamous cell carcinoma; lymphoepithelial carcinoma; basal cell carcinoma; pilomatrix carcinoma; transitional cell carcinoma; papillary transitional cell carcinoma; adenocarcinoma; gastrinoma, malignant; cholangiocarcinoma; hepatocellular carcinoma; combined hepatocellular carcinoma and cholangiocarcinoma; trabecular adenocarcinoma; adenoid cystic carcinoma; adenocarcinoma in adenomatous polyp; adenocarcinoma, familial polyposis coli; solid carcinoma; carcinoid tumor, malignant; branchiolo-alveolar adenocarcinoma; papillary adenocarcinoma; chromophobe carcinoma; acidophil carcinoma; oxyphilic adenocarcinoma; basophil carcinoma; clear cell adenocarcinoma; granular cell carcinoma; follicular adenocarcinoma; papillary and follicular adenocarcinoma; nonencapsulating sclerosing carcinoma; adrenal cortical carcinoma; endometroid carcinoma; skin appendage carcinoma; apocrine adenocarcinoma; sebaceous adenocarcinoma; ceruminous; adenocarcinoma; mucoepidermoid carcinoma; cystadenocarcinoma; papillary cystadenocarcinoma; papillary serous cystadenocarcinoma; mucinous cystadenocarcinoma; mucinous adenocarcinoma; signet ring cell carcinoma; infiltrating duct carcinoma; medullary carcinoma; lobular carcinoma; inflammatory carcinoma; paget's disease, mammary; acinar cell carcinoma; adenosquamous carcinoma; adenocarcinoma w / squamous metaplasia; thymoma, malignant; ovarian stromal tumor, malignant; thecoma, malignant; granulosa cell tumor, malignant; and roblastoma, malignant; Sertoli cell carcinoma; leydig cell tumor, malignant; lipid cell tumor, malignant; paraganglioma, malignant; extra-mammary paraganglioma, malignant; pheochromocytoma; glomangiosarcoma; malignant melanoma; amelanotic melanoma; superficial spreading melanoma; malig melanoma in giant pigmented nevus; epithelioid cell melanoma; blue nevus, malignant; sarcoma; fibrosarcoma; fibrous histiocytoma, malignant; myxosarcoma; liposarcoma; leiomyosarcoma; rhabdomyosarcoma; embryonal rhabdomyosarcoma; alveolar rhabdomyosarcoma; stromal sarcoma; mixed tumor, malignant; mullerian mixed tumor; nephroblastoma; hepatoblastoma; carcinosarcoma; mesenchymoma, malignant; brenner tumor, malignant; phyllodes tumor, malignant; synovial sarcoma; mesothelioma, malignant; dysgerminoma; embryonal carcinoma; teratoma, malignant; struma ovarii, malignant; choriocarcinoma; mesonephroma, malignant; hemangiosarcoma; hemangioendothelioma, malignant; kaposi's sarcoma; hemangiopericytoma, malignant; lymphangiosarcoma; osteosarcoma; juxtacortical osteosarcoma; chondrosarcoma; chondroblastoma, malignant; mesenchymal chondrosarcoma; giant cell tumor of bone; ewing's sarcoma; odontogenic tumor, malignant; ameloblastic odontosarcoma; ameloblastoma, malignant; ameloblastic fibrosarcoma; pinealoma, malignant; chordoma; glioma, malignant; ependymoma; astrocytoma; protoplasmic astrocytoma; fibrillary astrocytoma; astroblastoma; glioblastoma; oligodendroglioma; oligodendroblastoma; primitive neuroectodermal; cerebellar sarcoma; ganglioneuroblastoma; neuroblastoma; retinoblastoma; olfactory neurogenic tumor; meningioma, malignant; neurofibrosarcoma; neurilemmoma, malignant; granular cell tumor, malignant; malignant lymphoma; Hodgkin's disease; Hodgkin's lymphoma; paragranuloma; malignant lymphoma, small lymphocytic; malignant lymphoma, large cell, diffuse; malignant lymphoma, follicular; mycosis fungoides; other specified non-Hodgkin's lymphomas; malignant histiocytosis; multiple myeloma; mast cell sarcoma; immunoproliferative small intestinal disease; leukemia; lymphoid leukemia; plasma cell leukemia; erythroleukemia; lymphosarcoma cell leukemia; myeloid leukemia; basophilic leukemia; eosinophilic leukemia; monocytic leukemia; mast cell leukemia; megakaryoblastic leukemia; myeloid sarcoma; hairy cell leukemia; Lynch syndrome (known as hereditary nonpolyposis colorectal cancer (HNPCC) syndrome) and CMMRD (constitutional mismatch repair deficiency) syndrome. In some embodiments, the cancer is a metastatic cancer. In some embodiments, the cancer is a chemoresistant cancer.

[0029] In some embodiments, the subject suffers from acute myeloid leukemia. As used herein, the term “acute myeloid leukaemia" or “AML” has its general meaning in the art and refers to a cancer of the blood and bone marrow. In some embodiments, the AML is a chemoresistant AML and / or a metastatic AML.

[0030] In some embodiments, the subject suffers from breast cancer. As used herein, the term “breast cancer” or “BC” has its general meaning in the art and refers to a cancer that begins as a growth of cells in the breast tissue. In some embodiments, the AML is a chemoresistant breast cancer and / or a metastatic breast cancer.

[0031] Typically the active ingredient of the present invention (e.g. succinate dehydrogenase inhibitor) is combined with pharmaceutically acceptable excipients, and optionally sustained- release matrices, such as biodegradable polymers, to form pharmaceutical compositions. The term "Pharmaceutically" or "pharmaceutically acceptable" refers to molecular entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to a mammal, especially a human, as appropriate. A pharmaceutically acceptable carrier or excipient refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. The carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetables oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminium monostearate and gelatin. In the pharmaceutical compositions of the present invention, the active ingredients of the invention can be administered in a unit administration form, as a mixture with conventional pharmaceutical supports. Suitable unit administration forms comprise oral-route forms such as tablets, gel capsules, powders, granules and oral suspensions or solutions, sublingual and buccal administration forms, aerosols, implants, subcutaneous, transdermal, topical, intraperitoneal, intramuscular, intravenous, subdermal, transdermal, intrathecal and intranasal administration forms and rectal administration forms.

[0032] The invention will be further illustrated by the following figures and examples. However, these examples and figures should not be interpreted in any way as limiting the scope of the present invention.

[0033] FIGURES:

[0034] Figure 1. Viable cell number was assessed by Trypan blue staining on H9C2 rat cardiomyoblasts cell line for 24h with or without Doxorubicin (0.4pM) + Cytarabine (4pM) and malonate (lOOpM).

[0035] Figure 2. Viable cell number was assessed by Trypan blue staining on H9C2 rat cardiomyoblasts cell line for 24h with or without Doxorubicin (0.5pM) + Cyclophosphamide (lOOpM) and malonate (lOOpM). Figure 3. Assessment of left ventricular ejection fraction (LVEF) by echocardiography two weeks after treatments with cytarabine + Doxorubicin or Mai onate or combination of both. Welch ANOVA statistical test with Dunnett T3 Correction was used here. ** for pV<0.01 and *** for pVO.OOl.

[0036] Figure 4. Fibrosis area assessment with picrosirius red staining of heart sections two weeks after treatments with cytarabine + Doxorubicin or Malonate or combination of both. Welch ANOVA statistical test with Dunnett T3 Correction was used here. ** for pV<0.01 and *** for pVO.OOl.

[0037] Figure 5. Synergy assay on AML cell line M0LM14 treated for 24h with increasing dose of Doxorubicin + Cytarabine and malonate.

[0038] Figure 6. Annexin V assay on AML cell line M0LM14 treated for 24h with or without Doxorubicin (0.1 pM) + Cytarabine (IpM) and malonate (lOOpM).

[0039] Figure 7. Synergy assay on breast cancer cell line 4T1 treated for 24h with increasing dose of Doxorubicin + Cyclophosphamide and malonate.

[0040] Figure 8. Viable cell number was assessed by Trypan blue staining on 4T1 breast cancer cell line for 24h with or without Doxorubicin (1 pM) + Cyclophosphamide (ImM) and malonate (lOOpM).

[0041] Figure 9. Viable cell number was assessed by Trypan blue staining on H9C2 rat cardiomyoblasts cell line for 48h with or without Ibrutinib (lOpM) and malonate (lOOpM).

[0042] Figure 10. Viable cell number was assessed by Trypan blue staining on H9C2 rat cardiomyoblasts cell line for 24h with or without doxorubicin (0.4pM) + cytarabine (4pM) and Atpenin A5 (lOOnM).

[0043] Figure 11. Viable cell number was assessed by Trypan blue staining on H9C2 rat cardiomyoblasts cell line for 48h with or without Ibrutinib (lOpM) and Atpenin A5 (lOOnM). EXAMPLE 1:

[0044] Material & Methods

[0045] Viable H9C2 cell number with or without Doxorubicin + Cytarabine and malonate. H9C2 cell line was cultured in MEMa media (Gibco) supplemented with 10% FBS and 1% penicillin / streptomycin. Cells were plated at a density of 0.1 million cells per well in a 12 well plate and treated with or without Doxorubicin (0.4pM) + Cytarabine (4pM) and malonate (lOOpM) for 24 hours. Following treatments, cell counting was performed using a Mallassez glass slide counter after Trypan blue staining.

[0046] Viable H9C2 cell number with or without Doxorubicin + Cyclophosphamide and malonate. H9C2 cell line was cultured in MEMa media (Gibco) supplemented with 10% FBS and 1% penicillin / streptomycin. Cells were plated at a density of 0.1 million cells per well in a 12 well plate and treated with or without Doxorubicin (0.5pM) + Cyclophosphamide (lOOpM) and malonate (lOOpM) for 24 hours. Following treatments, cell counting was performed using a Mallassez glass slide counter after Trypan blue staining.

[0047] Viable H9C2 cell number with or without Ibrutinib and malonate. H9C2 cell line was cultured in MEMa media (Gibco) supplemented with 10% FBS and 1% penicillin / streptomycin. Cells were plated at a density of 0.1 million cells per well in a 12 well plate and treated with or without Ibrutinib (lOpM) and malonate (lOOpM) for 48 hours. Following treatments, cell counting was performed using a Mallassez glass slide counter after Trypan blue staining.

[0048] Assessment of left ventricular ejection fraction (LVEF) in mice after treatments with cytarabine + Doxorubicin or Malonate or combination of both. C57B1 / 6J mice received intravenous administration of doxorubicin (3 mg / kg) for three consecutive days along with intraperitoneal administration of cytarabine (100 mg / kg) for five consecutive days. Additionally, another group of mice was treated with intraperitoneal injections of dimethyl malonate (100 mg / kg) for three weeks, including the week of chemotherapy administration (total of 15 doses). Systolic cardiac function was evaluated using non-invasive echocardiography. The procedure involved lightly anesthetizing the mice with 1% isoflurane in air and placing them on a heating pad. Left ventricular dimensions were obtained via Time Movement mode acquisition from the parasternal short-axis view at the midventricular level using a Vevo2100 echograph and a 40 MHz transducer (M550; Fujifilm Visualsonics, Tokyo, Japan). Offline image analysis was performed using Vevolab software (Fujifilm Visualsonics).

[0049] Fibrosis area assessment in mice after treatments with cytarabine + Doxorubicin or Malonate or combination of both. Center section of the heart was slowly frozen in OCT (VWR, Radnor, PA, USA; #361603E) using an -80°C isopropanol bath. Briefly, hearts were transversely sectioned using a cryotome (Thermo NX50; Thermo Fisher Scientific) rinsed quickly in 1 change of acetic acid solution prior to mounting in 95% alcohol with 2 changes in Histoclear solution (Sigma-Aldrich) followed by mounting in synthetic resin. Slides were fixed in 4% PFA in 0.1 M sodium phosphate buffer pH 7.4 for 10 minutes before staining. Picrosirius red staining was performed to detect fibrosis according to the protocol of Junqueira et alu. Total fibrotic area was quantified by using NTH ImageJ software after scanning on a NanoZoomer (Hamamatsu Photonics, Shizuoka, Japan).

[0050] Synergy assay on AML cell line MOLM14 treated with increasing dose of Doxorubicin + Cytarabine and malonate. M0LM14 cell line was cultured in MEMa media (Gibco) supplemented with 10% FBS and 1% penicillin / streptomycin. Cells were plated at a density of 0.3 million per milliliter and treated with or without Doxorubicin (0.05 or 0.1) + Cytarabine (0.5 or 1 pM) and malonate (1, 10, 100, 500 and 1000 pM) for 24 hours. Following treatments, cell counting was performed using a Mallassez glass slide counter after Trypan blue staining. Cumulative index was calculated with CompuSyn software.

[0051] Annexin V assay on AML cell line MOLM14 treated with or without Doxorubicin + Cytarabine and malonate. M0LM14 cell line was cultured in MEMa media (Gibco) supplemented with 10% FBS and 1% penicillin / streptomycin. Cells were plated at a density of 0.3 million cells per milliliter and treated with or without Doxorubicin (0.1 pM) + Cytarabine (1 pM) and malonate (100 pM) for 24 hours. Subsequently, cells were washed and resuspended in annexin V binding buffer (BD Biosciences) in the presence of Annexin V APC (diluted at 1 / 200). Viable cells were then analyzed using aFortessa 10X flow cytometer with the assistance of positive and negative controls.

[0052] Synergy assay on breast cancer cell line 4T1 treated with increasing dose of Doxorubicin + Cyclophosphamide and malonate. 4T1 cell line was cultured in MEMa media (Gibco) supplemented with 10% FBS and 1% penicillin / streptomycin. Cells were plated at a density of 0.1 million cells per well in a 12 well plate and treated with or without Doxorubicin (0.5, 1 and 2 pM) + Cyclophosphamide (0.5, 1 and 2 mM) and malonate (10, 100, 500 and 1000 pM) for 24 hours. Following treatments, cell counting was performed using a Mallassez glass slide counter after Trypan blue staining. Cumulative index was calculated with CompuSyn software.

[0053] Viable 4T1 cell number with or without Doxorubicin + Cyclophosphamide and malonate. 4T1 cell line was cultured in MEMa media (Gibco) supplemented with 10% FBS and 1% penicillin / streptomycin. Cells were plated at a density of 0.1 million cells per well in a 12 well plate and treated with or without Doxorubicin (1 pM) + Cyclophosphamide (1 mM) and malonate (100 pM) for 24 hours. Following treatments, cell counting was performed using a Mallassez glass slide counter after Trypan blue staining.

[0054] Results

[0055] To evaluate the potential cardioprotective effect of succinate dehydrogenase inhibition against the cardiac toxicity of chemotherapies used to treat acute myeloid leukemia (AML) or breast cancer, we first tested its effect on in vitro cell viability. For AML, we examined the doxorubicin + cytarabine combination on rat H9C2 cardiomyoblastic cells, assessing viability at 24 hours. The decrease in viable cell count after 24 hours of treatment with this chemotherapy combination is almost entirely inhibited by adding malonate to the therapeutic combination (Figure 1).

[0056] In the context of breast cancer, we investigated the doxorubicin + cyclophosphamide combination and observed similar results in vitro (Figure 2).

[0057] To confirm the results of Figure 1 in vivo, we treated C57B1 / 6J mice with the same chemotherapy combination and demonstrated the cardioprotective effect of Malonate (here in the form of dimethyl-malonate to facilitate molecule entry into cells) by studying cardiac function through echocardiography and histology. Measurement of left ventricular ejection fraction highlighted the cardioprotective effect of malonate on the doxorubicin + cytarabine combination in vivo, with a return to normal values of this parameter 2 weeks after chemotherapy treatment (Figure 3). Histological examination of cardiac fibrosis revealed similar results, illustrating the antifibrotic effect of malonate in this model of chemotherapy- induced cardiotoxicity for AML treatment (Figure 4). Next, we sought to study the effect of malonate on the therapeutic resistance of cancer cells. It has been shown that AML cells are highly dependent on mitochondrial metabolism8'9, so we investigated whether the addition of malonate could sensitize the chemoresistant AML cell line M0LM14. The use of increasing doses of chemotherapy and malonate allowed us to conclude on the synergy of malonate and the chemotherapy combination in M0LM14 cells (Figure 5), illustrated at a fixed dose by an increase in the apoptotic population via an Annexin V assay measured by flow cytometry (Figure 6).

[0058] We demonstrated this same synergy in an aggressive metastatic murine breast cancer cell line 4T1 by combining malonate with doxorubicin and cyclophosphamide (Figure 7), illustrated at a fixed dose in Figure 8.

[0059] Finally and as demonstrated in Figure 9, Malonate significantly protected H9C2 cardiomyoblasts from ibrutinib-induced toxicity after 48 hours of treatment.

[0060] EXAMPLE 2:

[0061] Material & Methods

[0062] Viable H9C2 cell number with or without Doxorubicin + Cytarabine and atpenin A5. H9C2 cell line was cultured in MEMa media (Gibco) supplemented with 10% FBS and 1% penicillin / streptomycin. Cells were plated at a density of 0.1 million cells per well in a 12 well plate and treated with or without Doxorubicin (0.4pM) + Cytarabine (4pM) and atpenin A5 (lOOnM) for 24 hours. Following treatments, cell counting was performed using a Mallassez glass slide counter after Trypan blue staining.

[0063] Viable H9C2 cell number with or without Ibrutinib and atpenin A5. H9C2 cell line was cultured in MEMa media (Gibco) supplemented with 10% FBS and 1% penicillin / streptomycin. Cells were plated at a density of 0.1 million cells per well in a 12 well plate and treated with or without Ibrutinib (lOpM) and Atpenin A5 (lOOnM) for 48 hours. Following treatments, cell counting was performed using a Mallassez glass slide counter after Trypan blue staining.

[0064] Results

[0065] Atpenin A5 (AA5), a SDH inhibitor, significantly protected H9C2 cardiomyoblasts from intensive chemotherapy (iCT)-induced toxicity after 24 hours of treatment (Figure 10). Atpenin A5 also significantly protected H9C2 cardiomyoblasts from ibrutinib-induced toxicity after 48 hours of treatment (Figure 11). Taken together, these results confirm the relevance of targeting SDH in cardio-oncology.

[0066] CONCLUSION

[0067] The evidence provided by these experiments demonstrates the therapeutic effect of succinate dehydrogenase inhibition to counteract the cardiotoxicity of chemotherapies used in AML and breast cancer. This same combination also remarkably exhibits a chemosensitizing capacity of cancer cells. The use of Malonate in patients affected by these pathologies would therefore protect the heart from the toxic side effects of chemotherapy and eliminate the most resistant cells for an ever better risk-benefit ratio for these patients.

[0068] REFERENCES:

[0069] Throughout this application, various references describe the state of the art to which this invention pertains. The disclosures of these references are hereby incorporated by reference into the present disclosure.

[0070] 1. Herrmann, J. Adverse cardiac effects of cancer therapies: cardiotoxicity and arrhythmia. Nat Rev Cardiol 17, 474-502 (2020).

[0071] 2. Sturgeon, K. M. et al. A population-based study of cardiovascular disease mortality risk in US cancer patients. European Heart Journal 40, 3889-3897 (2019).

[0072] 3. Beck, T. C. et al. Cellular and Molecular Mechanisms of MEK1 Inhibitor- Induced Cardiotoxicity. JACC: CardioOncology 4, 535-548 (2022).

[0073] 4. Dickerson, T. et al. Hypertension and incident cardiovascular events following ibrutinib initiation. Blood 134, 1919-1928 (2019).

[0074] 5. Wallace, K. B., Sardao, V. A. & Oliveira, P. J. Mitochondrial Determinants of Doxorubicin-Induced Cardiomyopathy. Circulation Research 126, 926-941 (2020).

[0075] 6. Chouchani, E. T. et al. Ischaemic accumulation of succinate controls reperfusion injury through mitochondrial ROS. Nature 515, 431-435 (2014).

[0076] 7. Bae, J. et al. Malonate Promotes Adult Cardiomyocyte Proliferation and Heart Regeneration. Circulation 143, 1973-1986 (2021).

[0077] 8. Farge, T. et al. Chemotherapy-resistant human acute myeloid leukemia cells are not enriched for leukemic stem cells but require oxidative metabolism. Cancer Discovery 7, 716-735 (2017). 9. Bose, C. et al. Mitochondrial inhibitors circumvent adaptive resistance to venetoclax and cytarabine combination therapy in acute myeloid leukemia. Nature Cancer 1- 20 (2021) doi: 10.1038 / s43018-021-00264-y.

[0078] 10. Ojha, R. et al. Regulation of reverse electron transfer at mitochondrial complex I by unconventional Notch action in cancer stem cells. Developmental Cell 57, 260-276. e9 (2022).

[0079] 11. Erdem, A. et al. Inhibition of the succinyl dehydrogenase complex in acute myeloid leukemia leads to a lactate-fuelled respiratory metabolic vulnerability. Nat Commun 13, 2013 (2022). 12. Junqueira, L. C., Bignolas, G. & Brentani, R. R. Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections. Histochem J 11, 447-455 (1979).

Claims

CLAIMS:

1. A method for the treatment of chemotherapy-induced cardiotoxicity in a subj ect in need thereof comprising administering to said subject a therapeutically effective amount of a succinate dehydrogenase inhibitor.

2. The method according to claim 1, wherein the succinate dehydrogenase inhibitor is administrated simultaneously, separately or sequentially with at least one chemotherapeutic agent.

3. The method according to any of claims 1 or 2, wherein the treatment is a preventive treatment.

4. The method according to any of claims 1 to 3, wherein the at least one chemotherapeutic agent is an anthracycline.

5. The method according to any of claims 1 to 3, wherein the at least one chemotherapeutic agent is doxorubicin.

6. The method according to any of claims 1 to 3, wherein the at least one chemotherapeutic agent is a BTK inhibitor.

7. The method according to any of claims 1 to 3, wherein the at least one chemotherapeutic agent is Ibrutinib.

8. The method according to any of claims 1 to 3, wherein the at least one chemotherapeutic agent is an anthracycline and; an antimetabolite or an alkylating agent.

9. The method according to any of claims 1 to 3, wherein the at least one chemotherapeutic agent is doxorubicin and cyclophosphamide.

10. The method according to any of claims 1 to 3, wherein the at least one chemotherapeutic agent is doxorubicin and cytarabine.

11. The method according to any of claims 1 to 10, wherein the subject suffers from Acute Myeloid Leukemia (AML).

12. The method according to claim 11, wherein the AML is a chemoresistant AML.

13. The method according to any of claims 1 to 10, wherein the subject suffers from Breast Cancer (BC).

14. The method according to claim 13, wherein the breast cancer is a metastatic breast cancer.

15. A method for the treatment of a chemoresistant cancer in a subject in need thereof comprising administering to said subject a therapeutically effective amount of a succinate dehydrogenase inhibitor and at least one chemotherapeutic agent.

16. The method according to any of claims 1 to 15, wherein the succinate dehydrogenase inhibitor is malonate, disodium malonate or dimethyl malonate.

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