N-phenyl-quinoline-4-carboxamide compounds for use in the treatment of cancer

Abstract

The present invention pertains to the use of small molecules in the treatment of cancer. These compounds, which are small-molecule Furin inhibitors, show an ability to enhance sensitivity to immunotherapy and radiotherapy treatments, leading to tumor regression in mice and improved overall survival. Treatment of cancer cells with these compounds resulted in the elimination of the capacity for tumor cells and organoids to grow and invade. These results demonstrate the interest of using these small-molecule Furin inhibitors as therapeutic interventions for cancer patients.

Description

DescriptionTitle:N-PHENYL-QUINOLINE-4-CARBOXAMIDE COMPOUNDS FOR USE IN THE TREATMENT OF CANCERTechnical Field

[0001] The present invention pertains to the use of small molecules in the treatment of cancer.Backg ound Art

[0002] Tumor-reactive T cells are essential to antitumor immunity and are intricately regulated by a variety of coinhibitory and costimulatory immune checkpoints. This regulation is critical for ensuring an efficient immune response against tumors. Thereby, cancer immunotherapy aims to improve the survival and functionality of these T cells. One promising approach in this context is immune checkpoint blockade (ICB), which functions by interrupting pathways that suppress antitumor immunity, thus strengthening the body's capacity to fight against cancer cells1-6. ICB has revolutionized cancer treatments by proving effective even in advanced cases2’7-12-

[0003] Nevertheless, both primary and acquired resistance to ICB therapies persist as significant limitations13-15. Various studies indicate that elevated expression of several coinhibitory immune checkpoint receptors or ligands within the tumor microenvironment (TME) is linked to unfavorable treatment outcomes 16,17,17-19. Likewise, the lack of costimulatory immune checkpoints has also been linked to resistance to immunotherapy20-22. The majority of these regulatory factors or their inducer proteins, including growth factors, receptors, adhesion molecules necessitate the proprotein convertases (PC) Furin for their expression or activity2324.

[0004] Furin, also known as PCSK3, plays critical roles in both tumorigenesis and the modulation of immune responses23’25’26. Theoretically, Furin's involvement extends to all human cancers, affecting protein precursor activation and many mRNA expression across various cancer cell types23’25’26. Furin mediates the maturation of unprocessed substrates into bioactive molecules by cleaving specific basic amino acid motifs. These motifs are typically characterized as (K / R)-(X)n-(K / R);, where n can range from 0, 2, 4, or 6, and X represents any amino acid. In driving malignancy, proteins cleaved by Furin regulate critical aspects of cancer biology, such as cellular proliferation, survival, invasion, angiogenesis, metastasis, drug resistance, and immune evasion.27-30

[0005] Cumulative evidence supports Furin as an attractive therapeutic target in many cancers and other human diseases.23’25Silencing Furin expression or impeding its activity within diverse tumor models results in tumor regression, concomitant with a restructuring of the tumor microenvironment27’31. Previous investigations, have demonstrated that inhibiting Furin in cancer cells through cell transfection experiments and the utilization of protein- or peptide-based Furin inhibitors, such as a1 - PDX and Chloromethyl ketone (CMK)2932, effectively suppresses the malignant phenotype across diverse cancer cell types, impeding tumor growth in murine models25’30’33.

[0006] Numerous compounds with Furin-inhibitory activity have been identified, including both peptide-based and non-peptide compounds. Peptide-based molecules, such as d-Arg-based peptides, a1 -antitrypsin Portland51, and decanoyl-Arg-Val-Lys-Arg-chloromethylketone (dec-RVKR-Cmk)32, have been investigated in vitro for their ability to inhibit Furin activity. However, peptide- based inhibitors often face limitations such as susceptibility to degradation or poor absorption in the intestinal tract, thereby limiting their potential clinical utility52’53. Additionally, while several peptidomimetic drugs have been proposed and shown to inhibit Furin by binding to the enzyme's active site in vitro, these cyclic peptides have not demonstrated activity in cellular or in vivo models54. Small molecule-based competitive Furin inhibitors, such as Guanidinylated Aryl 2,5- Dideoxystreptamine Derivatives (GADDs)55, have also been developed. However, no preclinical studies have reported on the efficacy of such molecules.

[0007] Therefore, the development of small molecules with the capacity to directly bind to and impede Furin activity appears highly valuable.Summary

[0008] In this context, the inventors have identified a series of small molecules that inhibit Furin activity.

[0009] These compounds show an ability to enhance sensitivity to immunotherapy and radiotherapy treatments, leading to tumor regression in mice and improved overall survival. Treatment of cancer cells with these compounds resulted in the elimination of the capacity for tumor cells and organoids to grow and invade. These results demonstrate the interest of using these small-molecule Furin inhibitors as therapeutic interventions for cancer patients.

[0010] The first object of the invention is thus a compound of formula (I)whereinRi , R2, R3, R4, R5, Re, R7, Rs, R9, R10, R11 are independently selected from hydrogen, halogen, amine, O-s alkyl, C1-8 alkoxy, -NO2, -CN, optionally substituted phenyl, and optionally substituted heterocycle, for use in a method of treatment of cancer, said method comprising administering to a subject in need thereof an effective amount of compound of formula (I) or of a pharmaceutically acceptable salt thereof.

[0011] Another object of the invention is a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one other ingredient selected from a pharmaceutically acceptable excipient, diluent or carrier, for use in a method of treatment ofcancer.Detailed Description

[0012] A first object of the invention is a compound of formula (I)whereinRi , R2, R3, R4, R5, Re, R7, Rs, R9, R10, R11 are independently selected from hydrogen, halogen, amine, O-s alkyl, C1-8 alkoxy, -NO2, -CN, optionally substituted phenyl, and optionally substituted heterocycle, for use in a method of treatment of cancer, said method comprising administering to a subject in need thereof an effective amount of compound of formula (I) or of a pharmaceutically acceptable salt thereof.

[0013] The inventors demonstrated that compounds of formula (I) have a significative anti-tumor activity, which is associated with enhanced overall survival and infiltration of CD8+ cells in developed tumors. These compounds exhibit efficacy both in vitro and in vivo. In particular, they inhibit the proliferation and survival of colon cancer cells in 2D and 3D cultures, patient-derived colon organoids, and syngeneic mouse tumor models. Furthermore, in vivo experiments revealed that these compounds mediate CD8+ T cell infiltration, which correlates with reduced tumor growth and improved overall survival. Moreover, the inventors found that treatment of T cells with compounds of formula (I) led to the repression of PD-1 expression without affecting their activation and cytotoxic function.

[0014] The experiments led by the inventors demonstrate the pharmacological inhibition of Furin using the identified small molecules of formula (I), that exhibits promise in cancer, in particular in colon cancer models, demonstrating sustained effects in promoting tumor regression.

[0015] In the context of the invention, the term “halogen” refers to fluorine, chlorine, bromine, iodine.

[0016] In the context of the invention, the term “C1-8 alkyl” refers to C1-8 linear ( / .e., "straight-chain"), branched, or cyclic, saturated hydrocarbon chains, including for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, groups.

[0017] In the context of the invention, the term “alkoxy” refers to an alkyl ether group, i.e ., -(O-alkyl), where the alkyl is as defined above. In addition, “C1-8 alkoxy” may refer to alkoxy containing C1-8 alkyl, i.e., -(O-C1-8 alkyl), including for example, methoxy, ethoxy, propoxy, butoxy groups.

[0018] In the context of the invention, the term “optionally substituted phenyl” refers to is a cyclic group of atoms with the formula CeHs. The phenyl can be unsubstituted or substituted with at least one suitable substituent, for example with an optionally substituted Ci-Ce alkyl, or Ci-s alkoxy, or by -NO2, or -CN.

[0019] In the context of the invention, the term “optionally substituted heterocycle” refers to a cyclic compound that has atoms of at least two different elements as members of its ring(s). Preferably, the heterocycle is a five- or six-membered ring. Preferably, the heterocycle has from 1 to 3 heteroatoms selected from nitrogen, oxygen, and sulfur. The heterocycle may be an “heterocycloalkyl”, i.e. , a nonaromatic monocyclic or polycyclic ring comprising carbon and hydrogen atoms and at least one heteroatom, or may be an “heteroaromatic”, i.e., an aromatic ring containing at least one heteroatom as part of the aromatic ring. Examples of heterocycloalkyl groups include but without being limited to aziridinyl, pyrrolidinyl, pyrrolidino, piperidinyl, piperidino, piperazinyl, piperazino, morpholinyl, morpholino, thiomorpholinyl, thiomorpholino, tetrahydrofuranyl, tetrahydrothiofuranyl, tetrahydropyranyl, and pyranyl. Examples of heteroaromatic groups include but without being limited to pyridine, furan, thiophene, cytosine, and indole. The heterocycle can be unsubstituted or substituted with one or two suitable substituents, for example with an optionally substituted Ci-Ce alkyl, or C1-8 alkoxy, or by -NO2, or -CN.

[0020] In the context of the invention, the term “administration”, or a variant thereof (e.g., “administering”, “administered”) refers to the provision of an active agent or active ingredient, alone or as part of a pharmaceutically acceptable composition, to the subject or patient in whom / which the condition, symptom, or disease is to be treated, attenuated, visualised or diagnosed.

[0021] In the context of the invention, the terms “subject” or “patient” refers to a warm-blooded animal, more preferably a human, who / which is awaiting or receiving medical care or is or will be the object of a medical procedure.

[0022] The term “human” here refers to subjects of both genders and at any stage of development (i.e. neonate, infant, juvenile, adolescent, adult). In one embodiment, the human is an adolescent or adult, preferably an adult.

[0023] For the avoidance of doubt, references herein to "treatment" include references to curative, palliative and prophylactic treatment. A “treatment” aims at reversing, alleviating, inhibiting the progress of, or preventing the disorder or condition to which such term applies, or reversing, alleviating, inhibiting the progress of, or preventing one or more symptoms of the disorder or condition to which such term applies.

[0024] The term “cancer” herein refers to the physiological condition in subjects that is characterized by unregulated or dysregulated cell growth or death. The term "cancer" includes solid tumors and blood born tumors.

[0025] Typically, the compounds or compositions for use according to the present invention applies to various organs of cancer origin (such as breast, colon, gastric, rectum, pancreatic, lung, skin, headand neck, bladder, ovary, prostate, liver), and also to various cancer cell types (adenocarcinoma, squamous cell carcinoma, large cell cancer, melanoma, etc).

[0026] In a particular embodiment, the patient suffers from a solid cancer selected from the group consisting of skin cancer (e.g. melanoma, nonmelanoma skin cancer), colorectal cancer, adrenal cortical cancer, anal cancer, bile duct cancer (e.g. periphilar cancer, distal bile duct cancer, intrahepatic bile duct cancer), bladder cancer, bone cancer (e.g. osteoblastoma, osteosarcoma, chondrosarcoma, fibrosarcoma, malignant fibrous histiocytoma), sarcomas such as liposarcoma and soft-tissue sarcoma, brain and central nervous system cancer (e.g. meningioma, astocytoma, oligodendrogliomas, ependymoma, gliomas, medulloblastoma, ganglioglioma, germinoma, craniopharyngioma), breast cancer (e.g. ductal carcinoma in situ, infiltrating ductal carcinoma, infiltrating lobular carcinoma, lobular carcinoma in situ), cervical cancer, endometrial cancer (e.g. endometrial adenocarcinoma, adenocanthoma, papillary serous adnocarcinoma), esophagus cancer, gallbladder cancer (mucinous adenocarcinoma, small cell carcinoma), gastrointestinal carcinoid tumors (e.g. choriocarcinoma, chorioadenoma destruens), kidney cancer (e.g. renal cell cancer), laryngeal and hypopharyngeal cancer, liver cancer (e.g. hepatic adenoma, hepatocellular carcinoma), lung cancer (e.g. small cell lung cancer, non-small cell lung cancer), mesothelioma, nasal cavity and paranasal sinus cancer (e.g. esthesioneuroblastoma, midline granuloma), nasopharyngeal cancer, neuroblastoma, oral cavity and oropharyngeal cancer, ovarian cancer, pancreatic cancer, penile cancer, pituitary cancer, prostate cancer, retinoblastoma, rhabdomyosarcoma (e.g. embryonal rhabdomyosarcoma, alveolar rhabdomyosarcoma, pleomorphic rhabdomyosarcoma), salivary gland cancer, stomach cancer, testicular cancer (e.g. seminoma, nonseminoma germ cell cancer), thymus cancer, thyroid cancer (e.g. follicular carcinoma, anaplastic carcinoma, poorly differentiated carcinoma, medullary thyroid carcinoma), vaginal cancer, vulvar cancer, and uterine cancer (e.g. uterine leiomyosarcoma).

[0027] In the context of the invention, the terms “pharmaceutically acceptable” refer to ingredients of a pharmaceutical composition which are compatible with each other and not deleterious to the subject thereof.

[0028] In the context of the invention, the compounds represented by Formula (I) or (la) of the present invention may be used in the form of a pharmaceutically acceptable salt thereof. Particularly, the pharmaceutically acceptable salt may be an acid addition salt formed by a free acid. Here, acid addition salts may be obtained from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, and phosphorous acid; non-toxic organic acids such as aliphatic mono- and di-carboxylates, phenyl-substituted alkanoates, hydroxy alkanoates, and alkanedioates, aromatic acids, and aliphatic and aromatic sulfonic acids; and organic acids such as trifluoroacetic acid, acetate, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, and fumaric acid. The types of such pharmaceutically acceptable salts may include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, iodide, fluoride, acetate, propionate, decanoate, caprylate,acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1 ,4-dioate, hexane-1 ,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, p-hydroxyburyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1 -sulfonate, naphthalene-2- sulfonate, and mandelate. The acid addition salt may be prepared by a conventional method, for example, by dissolving a derivative of Formula (I) or (la) in an organic solvent such as methanol, ethanol, acetone, methylene chloride, or acetonitrile, adding an organic acid or inorganic acid, and filtering and drying the resulting precipitate, or prepared by distilling the solvent and excess acid under reduced pressure, dehydrating the resulting product and crystallizing the dehydrated product in an organic solvent. In addition, the pharmaceutically acceptable salt may be a salt or metal salt obtained using a base. As an example of the metal salt, an alkali metal or alkaline earth metal salt may be obtained by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering an undissolved compound salt, and evaporating and dehydrating the filtrate. As an alkali metal salt, a sodium, potassium, or calcium salt may be pharmaceutically acceptable. In addition, corresponding salts may be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

[0029] According to a particular embodiment, the compounds for use in a method of treatment of cancer according to the invention are selected from compounds of formula (Ia1 ), (Ia2) or (Ia3)whereinRi , R3, R4, R5, Re, R7, Rs, R9, R10, R11 , R12, R13, R14, R15, R16, R17, Ris, R19, R20 are independently selected from hydrogen, halogen, amine, Ci-s alkyl, Ci-s alkoxy, -NO2, -CN, optionally substituted phenyl, and optionally substituted heterocycle.

[0030] According to a particular embodiment, the compounds for use in a method of treatment of cancer according to the invention are selected from compounds of formula (la) or (lb)whereinR1 , R3, R4, R5, Re, R7, Rs, R9, R10, R11 , R12, R13, R14, R15, R16, R17, Ris, R19, R20 are independently selected from hydrogen, halogen, amine, Ci-s alkyl, Ci-s alkoxy, -NO2, -CN, optionally substituted phenyl, and optionally substituted heterocycle.

[0031] In another embodiment, the compounds for use in a method of treatment of cancer according to the invention are selected from

[0032] The inventors that the compounds of formula (I) can be advantageously used in combination with other anticancer therapy. Indeed, the experiments they carried out showed that combining small molecules Furin inhibitors of formula (I) with anti-PD-1 therapy and radiotherapy elicited superior responses compared to single therapies alone, supporting their hypothesis that anti-PD-1 therapy or radiotherapy, when combined with Furin inhibition, operate through distinct yet complementary mechanisms.

[0033] Indeed, while chemotherapy and radiation have been utilized to trigger an antitumor immune response by inducing tumor cell death, these treatments often coincide with T-cell dysfunction and toxicity. Therefore, combining them with small molecules Furin inhibitors allows to mitigate these side effects by reducing T-cell exhaustion. The inventor’s study furnishes molecular and preclinical evidence advocating for the catalytic inhibition of Furin as a supplementary therapy alongside anti- PD-1 treatment or radiotherapy.

[0034] Thus, according to a particular embodiment, the compounds for use in a method of treatment of cancer according to the invention are used in combination with at least one other anticancer therapy.

[0035] In particular, said other anticancer therapy can be selected from chemotherapy, targeted therapy, immunotherapy or radiation therapy.

[0036] “Chemotherapy” refers to chemical compounds that are effective in inhibiting tumor growth. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, trietylenephosphoramide, triethylenethiophosphaorarnide and trimethylolomelamine; acetogenins (especially bullatacin and bullatacinone); a camptothecin (including irinotecan and topotecan); bryostatin; callystatin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogues, KW- 2189 and CBI-TMI); eleutherobin; pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estrarnustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimus tine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as the enediyne antibiotics (e.g. calicheamicin, especially calicheamicin (1 1 and calicheamicin 21 1 , see, e.g., Agnew Chem Inti. Ed. Engl. 33:183-186 (1994); dynemicin, including dynemicin A; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antiobiotic chromomophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, canninomycin, carzinophilin, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idanrbicin, marcellomycin, mitomycins, mycophenolic acid, nogalarnycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin,streptomgrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophospharnide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfornithine; elliptinium acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidamol; nitracrine; pento statin; phenamet; pirarubicin; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK®; razoxane; rhizoxin; sizofiran; spirogennanium; tenuazonic acid; triaziquone; 2,2',2"-trichlorotriethylarnine; trichothecenes (especially T-2 toxin, verracurin A, roridinA and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobromtol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, e.g. paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, N.].) and docetaxel (TAXOTERE®, Rhone-Poulenc Rorer, Antony, France); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin, carboplatin oxaloplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; CPT-1 1 ; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoic acid; capecitabine; and pharmacally acceptable salts, acids or derivatives of any of the above. Also included in this definition are antihormonal agents that act to regulate or inhibit honnone action on tumors such as anti-estrogens including for example tamoxifen, raloxifene, aromatase inhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene, LY1 17018, onapristone, and toremifene (Fareston); and anti-androgens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptable salts, acids or derivatives of any of the above.

[0037] “Targeted therapy” refer to agents that act by blocking the growth of cancer cells by interfering with specific targeted molecules needed for carcinogenesis and tumor growth. Most targeted therapies are either small-molecule drugs or monoclonal antibodies. It is noteworthy that some targeted therapies can qualify as immunotherapeutic agents and / or chemotherapeutic agents. Examples of targeted therapies include Bortezomib, Braf inhibitors such as vemurafenib and dabrafenib, Cobimetinib, Imatinib, Gefitinib, Erlotinib Sorafenib, Sunitinib, Dasatinib, Lapatinib, Nilotinib, tamoxifen, janus kinase inhibitors such as Tofacitinib, ALK inhibitors such as Crizotinib, Bcl- 2 inhibitors such as Venetoclax, Obatoclax, navitoclax, and gossypol, PARP inhibitors such as olaparib, rucaparib, niraparib and talazoparib, PI3K inhibitors such as perifosine, Apatinib, Zoptarelin doxorubicin, MEK inhibitors such as trametinib, CDK inhibitors, Hsp90 inhibitors, Hedgehog pathway inhibitors such as vismodegib and sonidegib, salinomycin VAL-083, Vintafolide, Temsirolimus, Everolimus, Vemurafenib, Trametinib, Dabrafenib, monoclonal antibodies including Pembrolizumab, Rituximab, Alemtuzumab, Cetuximab, Panitumumab, Bevacizumab and Ipilimumab.

[0038] “Immunotherapy” refers to a compound, composition or treatment that indirectly or directly enhances, stimulates or increases the body's immune response against cancer cells and / or that decreases the side effects of other anticancer therapies. Immunotherapy is thus a therapy that directly or indirectly stimulates or enhances the immune system's responses to cancer cells and / or lessens the side effects that may have been caused by other anti-cancer agents. Immunotherapy is also referred to in the art as immunologic therapy, biological therapy, biological response modifier therapy and biotherapy. Examples of common immunotherapeutic agents known in the art include, but are not limited to, anti-PD-1 , anti-PDL-1 , anti-CTLA4 immunotherapies, cytokines, cancer vaccines, monoclonal antibodies and non-cytokine adjuvants. Alternatively, the immunotherapeutic treatment may consist of administering the patient with an amount of immune cells (T cells, NK, cells, dendritic cells, B cells...).

[0039] According to a particular embodiment, the compounds for use in a method of treatment of cancer according to the invention are used in the treatment of cancer selected from colon cancer, pancreatic cancer, breast cancer, liver cancer, leukemia, and prostate cancer.

[0040] According to another embodiment, the compounds for use in a method of treatment of cancer according to the invention are used in a method comprising administering to a subject in need thereof, said subject being a human, an effective amount of compound of formula (I) or of a pharmaceutically acceptable salt thereof.

[0041] Another object of the invention is a pharmaceutical composition comprising a compound of formula (I)whereinRi , R2, R3, R4, R5, Re, R7, Rs, R9, R10, R11 are independently selected from hydrogen, halogen, amine, C1-8 alkyl, C1-8 alkoxy, -NO2, -CN, optionally substituted phenyl, and optionally substituted heterocycle, or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient, for use in a method of treatment of cancer.

[0042] The term “excipient” as used herein means a substance formulated alongside the active agent or active ingredient in a pharmaceutical composition or medicament. Acceptable excipients for therapeutic use are well known in the pharmaceutical art. The choice of excipient can be selected with regard to the intended route of administration and standard pharmaceutical practice. The excipient must be acceptable in the sense of being not deleterious to the recipient thereof. The atleast one pharmaceutically acceptable excipient may be for example, a binder, a diluent, a carrier, a lubricant, a disintegrator, a wetting agent, a dispersing agent, a suspending agent, and the like.

[0043] Another object of the invention is a pharmaceutical composition comprising a compound selected from compounds of formula (la) or (lb)whereinRi , R3, R4, R5, Re, R7, Rs, R9, R10, R11 , R12, R13, R14, R15, R16, R17, Ris, R19, R20 are independently selected from hydrogen, halogen, amine, C1-8 alkyl, C1-8 alkoxy, -NO2, -CN, optionally substituted phenyl, and optionally substituted heterocycle, or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient, for use in a method of treatment of cancer.

[0044] According to a particular embodiment, the pharmaceutical composition for use in a method of treatment of cancer according to the invention comprises a compound selected from compounds of formula (Ia1 ), (Ia2) or (Ia3)whereinRi , R3, R4, R5, Re, R7, Rs, R9, R10, R11 , R12, R13, R14, R15, R16, R17, Ris, R19, R20 are independently selected from hydrogen, halogen, amine, C1-8 alkyl, C1-8 alkoxy, -NO2, -CN, optionally substituted phenyl, and optionally substituted heterocycle, or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient.

[0045] Another object of the invention is a pharmaceutical composition comprising a compound selected fromor a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient, for use in a method of treatment of cancer.

[0046] According to a particular embodiment, the compound or composition for use in a method of treatment of cancer according to the invention can be administered orally, enterally, intravenously, or intramuscularly.Brief Description of Drawings

[0047] Fig. 1

[0048] [Fig. 1 ] Furin Small Molecule Inhibitors Decrease Tumor Growth and Enhance Overall Survival. In vivo studies and analysis conducted in syngeneic mice inoculated subcutaneously with CT-26 and MC-38 cells (1 .106cells / injection). Mice were administered compounds intraperitoneally at 10mg / kg 3 to 5 times per week. Analysis of tumor growth and survival following injections of CT- 26 (B, R) and MC-38 (D, T) cells and treatment with compounds I0, and 113, respectively. Survival analysis of mice injected with CT-26 (C, S). Data are presented as mean ± SEM.*, P < 0.05, **, P < 0.01 , ***, P < 0.001 , “**, P < 0.0001 , determined by 2-way ANOVA test with Sidak’s multiple comparison test or log-rank test.Fig. 2

[0049] [Fig. 2] Compounds I0, and 113 Inhibit Cell Growth, Cell Invasion, and Enhance Cell Death and Intratumoral Cytotoxic T Cell Infiltration. Flow cytometry analysis and quantification of cell proliferation from tumors of five different colon cancer patients. Data are presented as mean ± SEM. *, P < 0.05, **, P < 0.01 , ***, P < 0.001 , P < 0.0001 , determined by 2-way ANOVA test with Sidak’s multiple comparison test or log-rank test or Wilcoxon matched-pairs signed rank test.

[0050] Fig. 3

[0051] [Fig. 3] A, Flow cytometry histogram of CFSE marker in controls and indicated compounds- treated cells-derived organoids. B, Flow cytometry analysis of percentage of cell proliferation. Data are shown as mean ± SEM. n = 5-10 / group. *, P < 0.05, **, P < 0,01 , ***, P < 0,001 , *“*, P < 0,0001 was determined by 2-way ANOVA test with Sidak’s multiple comparison test or log-rank test or Wilcoxon matched-pairs signed rank test.Fig. 4

[0052] [Fig. 4] Compounds I0, and 113 Inhibit Cell Growth, Cell Invasion, and Enhance Cell Death and Intratumoral Cytotoxic T Cell Infiltration. Representative microscopy images of organoids derived from CT-26 cells treated with or without compound I0, labeled with EB. Scale bar, 200pm. Cell death quantification is represented by EB intensity staining in the corresponding bar graphs. Data are presented as mean ± SEM. *, P < 0.05, **, P < 0.01 , ***, P < 0.001 , ****, P < 0.0001 , determined by 2-way ANOVA test with Sidak’s multiple comparison test or log-rank test or Wilcoxon matched-pairs signed rank test.

[0053] Fig. 5

[0054] [Fig. 5] I0 and 113 compounds induced cell death in organoids. Representative invert microscopy images of organoids from colon cancer line (CT-26) treated or not with I0 and 113respectively marked with EB and the corresponding analysis. Each red dot represents a dead cell. Scale bar, 200pm.Fig. 6

[0055] [Fig. 6] Compounds I0 and 113 Inhibit Cell Growth, Cell Invasion, and Enhance Cell Death and Intratumoral Cytotoxic T Cell Infiltration. Representative microscopy images of organoids from the colon cancer line (CT-26) treated with or without compounds I0 and 113, respectively, embedded in a collagen matrix. The bar graph indicates quantification of invasion area measured using the Fiji Macro analysis program. Scale bar, 200pm.Fig. 7

[0056] [Fig. 7] Compounds I0 and 113 Inhibit Cell Growth, Cell Invasion, and Enhance Cell Death and Intratumoral Cytotoxic T Cell Infiltration. L, Representative epifluorescence microscopy images and quantification of CD8+ T cells immunofluorescence within tumors derived from CT-26 injected mice and treated with compounds I0 and 113. Scale bar, 100pm. M, Tumor growth at the conclusion of the corresponding experiments on mice with colon tumors. Three to five independent experiments were conducted. Data are presented as mean ± SEM. *, P < 0.05, **, P < 0.01 , ***, P < 0.001 , “**, P < 0.0001 , determined by 2-way ANOVA test with Sidak’s multiple comparison test or log-rank test or Wilcoxon matched-pairs signed rank test.Fig. 8

[0057] [Fig. 8] Synergistic Effects of Furin Inhibition by Compound I0 with Immunotherapy and Radiotherapy to Constrain Tumor Growth in Mice and Suppress Cancer Cell Patient Growth. D-E, Analysis of tumor growth following CT-26 injection and treatment with compound I0 and / or anti- PD1 .F, Survival analysis of mice treated accordingly. G-H, Assessment of tumor growth post-CT-26 injection and treatment with compound I0 and / or ionizing radiation (IR). I, Corresponding survival analysis of the treated mice. K, N, Immunofluorescence analysis and quantification of CD8+ T cells from frozen sections derived from CT-26 injected mice treated with Compound I0 and anti-PD1 (K) or irradiated (N). L, O, Evaluation of tumor growth in mice injected with CT-26 and treated with compound I0 and anti-PD1 (L) or irradiation (O) compared to controls. Scale bar, 100pm. Data are presented as mean ± SEM.*, P < 0.05, **, P < 0.01 , ***, P < 0.001 , P < 0.0001 , determined by 2-way ANOVA test with Sidak’s multiple comparison test or log-rank test.Fig. 9

[0058] [Fig. 9] Sensitization of tumors to radiotherapy by furin silencing. B-C, Analysis of tumor growth following injection of MC-38 cells and irradiation treatment. D, Survival analysis of mice subjected to the corresponding treatments. Data are presented as mean ± SEM from 3 independent experiments.*, P < 0.05, ***, P < 0.001 , ****, P < 0.0001 , determined by 2-way ANOVA test with Sidak’s multiple comparison test or log-rank test.ExamplesMaterial and methodsReaqents and antibodiesThe antibodies used in this study include anti-human-PE-anti-PD-1 (130-120-382, Miltenyi Biotec), anti-human-PE-anti-CD69 Ab (IM1943U, Beckman Coulter), anti-mouse-APC / Fire750-anti-PD-1 (#135239, BioLegend), PE / Cya7-anti-CD8a (#100721 , BioLegend), and AF700-anti-CD69 (#104539, BioLegend). Additionally, anti-mouse CD8 (Ab217344, Abeam, 1 :500), anti-Ki67 (#9129T, Cell Signaling, dilution 1 :200), and anti-c-caspase 3 (#96617, Cell Signaling, dilution 1 :200) antibodies were utilized. The secondary antibody anti-rabbit (#71 1 -585-152, dilution 1 :300) was obtained from Jackson ImmunoResearch. All powders and reagents used in the study were acquired from Sigma. The compounds I0, 113, 11 and I4, used in the research, were purchased from commercial suppliers (ChemBridge, San Diego, USA) or MolPort (Riga, Latvia) and were dissolved in DMSO.Tumor samples, peripheral blood mononuclear cells (PBMCs), and T cells were obtained from colorectal cancer patients with approval from the Ethics Committee at Institut Bergonie in Bordeaux, France. All patients provided signed informed consent, and the studies adhered to Good Clinical Practice guidelines and the Declaration of Helsinki. Tissue specimens were promptly placed on ice and snap-frozen in liquid nitrogen after surgery for subsequent analysis or for use in organoid generation, as previously described 73. In additional experiments, either organoids or tissue samples sourced from patients were used to generate a single-cell suspension utilizing the tumor dissociation kit from Miltenyi Biotech (Bergisch Gladbach, Germany). PBMCs obtained from healthy donors were employed directly for protein expression analysis or underwent T-cell isolation utilizing the Pan T-cell isolation kit, following the manufacturer's instructions provided by Miltenyi Biotec.Cell culture and organoid generationThe HT29 human colon carcinoma cell line, along with the murine colon carcinoma cell lines CT26 and MC38, and the Jurkat human T-cell line, were obtained and authenticated from ATCC. Following procurement, each cell line underwent two passages before being stored or utilized in both in vitro and in vivo experiments. Culturing of the cells was carried out in complete media, utilizing either DMEM or RPMI 1640 (Gibco). Prior to experimentation, all cell lines were verified to be free from Mycoplasma contamination. To ensure Mycoplasma absence, the cells underwent a two-monthMycoplasma elimination process, which involved culturing them in ciprofloxacin. This process was confirmed by PCR analysis before their utilization in experiments. For the generation of CT26, HT- 29, and MC38 organoids, cells (5x103) were cultured in DMEM / F-12 supplemented with N2 and B27 supplements (Thermo Fisher Scientific), along with growth factors [20 ng / mL basic fibroblast growth factor (p-FGF) and 20 ng / mL epidermal growth factor (EGF) (Sigma)], and 0.4% sterile methylcellulose. This culture was conducted in a 96-well round-bottom plate, facilitating the formation of a single homogeneous organoid of cells.Mouse modelsThe research animals were housed at the University of Bordeaux in a temperature-controlled environment. Approval for all experimental procedures was obtained from the Institutional Animal Care and Use Committee at the University of Bordeaux, and these procedures were conducted under the supervision of trained veterinarians. BALB / c and C57BL6 mice aged 5 to 8 weeks were subcutaneously inoculated in the right flank with 1 x106 CT26 or MC38 colon carcinoma cells. Upon reaching tumor sizes of 25-50 mm3, the animals were randomly assigned to either the vehicle group (1 % DMSO in PBS) or the I0, 113, 11 and I4 compounds group (10 mg / kg intraperitoneally), with group sizes ranging from 7 to 10 mice for each experiment. In additional tests, mice received treatment with selected compounds (10 mg / kg intraperitoneally) combined with anti-PD-1 (5 mg / kg) or radiotherapy (2 Gy). Simultaneously, 6-8-week-old CD4cre-furflox / flox mice, characterized by T-cell-specific Furin deficiency 43,44, were subcutaneously injected with syngeneic MC-38 cells and subjected to radiotherapy or left untreated. Tumor progression was monitored three times weekly, and mice were euthanized at the end of the experiments. Tumor volume was calculated using the formula volume (mm3) = length [mm] x (width [mm])2 x 0.5216. In specific experiments, established tumors were dissociated for T-cell isolation following the manufacturer's instructions (Miltenyi Biotec).Cell proliferation assayTo evaluate the proliferation rates of both 2D cells and 3D organoids, cells and organoids were treated with an 8 pM CarboxyFluorescein Succinimidyl Ester (CFSE) marker (BioLegend) for 20 minutes. After thorough washing, the cells and organoids were exposed to the specified compounds. Following the incubation period, cells were rinsed with PBS, detached using 0.25% Trypsin, and subsequently suspended in PBS for analysis using flow cytometry (C6 Accuri, BD Biosciences).Cell viability assaysCell viability was evaluated through the MTT assay, as previously described73. Following exposure to the specified compounds for 24 and 48 hours, cells were incubated in an MTT solution (0.5 mg / mL) for 2 hours at 37°C. The absorbance was subsequently measured at 570 nm using an ELISA plate reader. For organoids, viability was assessed using 4 pM ethidium homodimer-1 (EB) from Invitrogen. Briefly, after treating organoids with the indicated compounds at concentrations of 10 or 50 pM, or with a vehicle solution, EB diluted in DPBS was added to the organoid wells for 1 hour at 37°C. The organoids were then imaged using a Nikon inverted microscope. Dead cells, visualized in red with the EB dye, were quantified using the Fiji Macro analysis program.Wound-healing assayCancer cells were plated in 6-well plates at a density of 1 x 105cells per well, leading to the formation of confluent monolayers. These monolayers were then deprived of serum overnight. Subsequently, straight wounds were introduced using 200-pl pipette tips, followed by washing with medium to clear away cell debris. The wounded monolayers were then exposed to medium containing 5% FBS along with treatment of 10 pM of the specified compounds or vehicle. Wound images were captured at 0 and 24 hours, and the area lacking cells within the wounds was quantified using Fiji software.Organoids collagen invasion assavFor the organoids collagen invasion assay, organoids were first washed twice in PBS and then mixed with 1 mg / mL rat tail Type I Collagen (Sigma). Subsequently, they were individually incubated in separate wells of a 96-well plate, either in the presence or absence of the specified compounds. All images were captured under consistent settings, and the invasion of collagen type I was quantified by subtracting the total area from the central area, using the Fiji Macro analysis program, as detailed previously73.Syngeneic mouse tumors obtained from CT26 and MC-38 cells were harvested, embedded in OCT, and stored at -80°C until use. Tissue sections underwent fixation in 4% PFA for 10 minutes, followed by three 5-minute washes in PBS and a 20-minute incubation in TBS-Tween 0.02%. After additional washes, the tissue sections were incubated in a blocking solution containing 5% bovine serum albumin (BSA; Euromedex) for 1 hour at room temperature (RT). Subsequently, sections were exposed overnight at 4°C to the primary antibody anti-mouse CD8 (Ab217344, Abeam) at a dilution of 1 :500 in PBS-0.1 % BSA. The sections were then treated with the appropriate fluorophore- conjugated secondary antibody at a 1 :500 dilution (Fluoprobes) for 1 hour at RT. To reduce tissue autofluorescence, sections were washed with Sudan Black and subjected to a series of decreasing ethanol solutions before DAPI labeling. Nuclei were counterstained with DAPI. Fluorophore signals were captured using an inverted microscope (Nikon), and the acquired images were analyzed using NIH Imaged.T cell activationActivation of TCR signaling in PBMCs, isolated T cells, and Jurkat T cells was conducted as previously described 74, utilizing either 5 pg / ml plate-bound anti-CD3 (clone OKT3, #317302, BioLegend) or anti-CD3 and anti-CD28 beads (T cell TransAct, Miltenyi Biotec). The activation period lasted for 24 hours. Treatment with I0, 113, 11 and I4 compounds occurred prior to TCR activation for 24 hours, after which cells were harvested for flow cytometry analyses, RNA / protein extraction, or cytotoxicity assays.Statistical analysis and visualizationThe statistical analyses were performed using Prism software (GraphPad). Various tests, including the Student t-test, Pearson correlation coefficient, Spearman’s rank correlation coefficient, andKaplan-Meier analysis (log-rank test), were employed. A significance level of P < 0.05 was considered, and the experimental data were presented as the mean ± standard error of the mean.Results1 . Tumor growth repressionTo assess the impact of the identified small molecule Furin inhibitors of formula (I) on tumor growth, syngeneic mouse hosts were subcutaneously (s.c.) inoculated with colon cancer cells CT-26 and MC-38. Subsequently, mice were randomly assigned to either a control or treatment group. The treatment group received an intraperitoneal dose of 10 mg / kg / compound when tumors reached a size of 25-50 mm3.The tested I0, 113 compounds exhibited a significant reduction in tumor growth induced by CT-26 (Fig. 2B, R) and MC-38 (Fig. 2D, T). This decrease in tumor growth correlated with an enhanced overall survival of treated mice (Fig. 2C, S).2. Inhibitory effects on cell proliferation, viability, and invasionThe inventors sought to determine whether the inhibited tumor growth in mice mediated by I0 and 113 is linked to a repression of these biological processes. They first assayed these compounds capability for inducing cell growth repression in the colon cancer cell line CT-26, MC-38 and HT-29 cells and derived organoids. By analyzing the fluorescence signal of CFSE, they found that these cells-derived organoids show different sensitivity to I0 and 113 compounds.They next evaluated I0 directly on cancer cells derived from colon cancer patients using CFSE. As illustrated in Fig. 3B, I0 (10 pM) reduced the number of proliferating cells by up to 50%, as assessed by CFSE assay.The use of the MTT assay revealed that compounds I0 and 113 (10 pM) significantly reduced the cell viability of CT-26, MC-38, and HT-29 cells (Supplementary Fig. S5). When using organoids derived from these cells, all compounds promoted CT-26 cell death, as assessed by the dead cell-permeable red fluorescent dye Ethidium homodimer-1 (Fig. 3C for CT-26, Supplementary Fig. S6 for MC-38 and HT-29).Next, the inventors investigated the impact of the inhibitors on the invasion of cancer cell-derived organoids in vitro, employing a 3D model of collagen cancer invasion. Notably, compounds I0 and 113 emerged as the primary inhibitors of collagen cell invasion (Fig. 3G-J). Additionally, the wound healing assay demonstrated a diminished motility potential of the cancer cells, as evidenced by delayed wound healing observed 24 hours after the scratch in the monolayer culture of cancer cells.3. Promotion of enhanced intratumoral infiltration of cytotoxic T cells.The inventors proceeded to quantify the presence of intratumoral cytotoxic T cells (CTLs) in mice under both control conditions and treatment with Furin inhibitors. Subcutaneous implantation of CT- 26 cells into syngeneic mouse hosts was performed, with mice randomly allocated into control or IO and 113-treated groups.The infiltration of CD8+ cells was then assessed within the developed tumors. Immunofluorescence staining revealed a significant increase in the number of infiltrating CD8+ cells in mice treated with all compounds (Fig. 3L), that associates the suppression of tumor growth compared to control mice tumors (Fig. 3M).4. Enhanced synergistic impact of immunotherapy and radiotherapy on tumor growth restraint by compound IOThe inventors assessed the potential synergistic effect of combining IO with anti-PD-1 in impeding tumor progression. CT-26 cells were implanted via intracutaneous injection, and mice were treated. While IO (10 mg / kg) notably reduced tumor growth, anti-PD-1 at a concentration of 100 pg / mouse didn't produce a significant reduction in tumor growth when administered alone. However, the combination treatment demonstrated a more pronounced limitation of tumor growth by day 25 (Fig. 4D-E). Furthermore, the combined use of IO and anti-PD-1 significantly increased the overall survival of treated mice (Fig. 4F).To explore whether this combinatorial effect could extend to other therapies, such as radiotherapy (I R), tumor cells were introduced via subcutaneous injection, and mice were randomized into control, IO-treated, and / or irradiated groups. The treatment groups received IO (10 mg / kg) or irradiation (IR, 2 Gy) alone or in combination. While IR led to a reduction in tumor growth, the combination treatment induced a more substantial reduction in tumor growth on day 25 (Fig. 4G-H). Additionally, the combination treatment increased overall survival compared to all other groups (Fig. 4I). Subsequently, the inventors evaluated whether I0, combined with checkpoint immunotherapy and ionizing radiation (IR), could further enhance CD8+ infiltration within tumors induced in mice. Thereby, mice were injected with CT-26 and were treated, either with I0 and anti-PD-1 or I0 and IR. While I0, anti-PD-1 , or IR individually led to a notable enhancement in CD8+ infiltration within tumors (approximately 4.36-fold, 5.16-fold, and 2.12-fold, respectively), the combined treatments of IO / anti- PD-1 (Fig. 4K) and I0+IR (Fig. 4N) demonstrated even greater increases in CD8+ infiltration (approximately 5.69-fold and 7.20-fold, respectively). This effect was observed across all mice tumors that exhibited reduced growth following these combination treatments (Fig. 4L, O).5. Silencing of Furin in murine T cells suppresses tumor growth and improves mice survival.To further confirm the observed effect of Furin repression on tumor growth and its influence on T cells mediated by I0, the inventors utilized mice lacking Furin specifically in T cells, known as CD4cre- furflox / flox (KO CD4 / Furin) mice43’44. MC-38 colon cancer cells were introduced into both KO CD4 / Furin and control syngeneic mice. During these experiments, they also examined the combinedeffect of T cell deficiency and IR in mice with MC-38 tumors. Consequently, the mice were categorized into non-irradiated and irradiated groups. The absence of Furin in T cells of KO CD4 / Furin mice resulted in a delayed growth of MC38 tumors (Fig 7B-C) and an improvement in overall survival (Fig 7D). Treatment with IR (2 Gy) induced a more pronounced reduction in tumor growth by day 25 (Fig 7B-C), leading to prolonged survival of the host mice (Fig. 7D). These findings indicate that the genetic repression of Furin, or its decreased activity in T cells, mediated by IO treatment, synergistically inhibits tumor growth and enhances overall survival when utilized in combination with other therapies.Citation List

[0059] 1 . Blank, C., Brown, I., Peterson, A.C., Spiotto, M., Iwai, Y., Honjo, T., and Gajewski, T.F. (2004). PD-L1 / B7H-1 inhibits the effector phase of tumor rejection by T cell receptor (TCR) transgenic CD8+ T cells. Cancer Res. 64, 1 140-1 145. 10.1 158 / 0008-5472.can-03-3259.2. Larkin, J., Chiarion-Sileni, V., Gonzalez, R., Grob, J.J., Cowey, C.L., Lao, C.D., Schadendorf, D., Dummer, R., Smylie, M., Rutkowski, P., et al. (2015). Combined Nivolumab and Ipilimumab or Monotherapy in Untreated Melanoma. N. Engl. J. Med. 373, 23-34. 10.1056 / NEJMoa1504030.3. Curiel, T.J., Wei, S., Dong, FL, Alvarez, X., Cheng, P., Mottram, P., Krzysiek, R., Knutson, K.L., Daniel, B., Zimmermann, M.C., et al. (2003). Blockade of B7-H1 improves myeloid dendritic cell-mediated antitumor immunity. Nat. Med. 9, 562-567. 10.1038 / nm863.4. Sierro, S.R., Donda, A., Perret, R., Guillaume, P., Yagita, FL, Levy, F., and Romero, P. (201 1 ). Combination of lentivector immunization and low-dose chemotherapy or PD-1 / PD-L1 blocking primes self-reactive T cells and induces anti-tumor immunity. Eur. J. Immunol. 41, 2217- 2228. 10.1002 / eji.201041235.5. Winograd, R., Byrne, K.T., Evans, R.A., Odorizzi, P.M., Meyer, A.R.L., Bajor, D.L., Clendenin, C., Stanger, B.Z., Furth, E.E., Wherry, E.J., et al. (2015). Induction of T-cell Immunity Overcomes Complete Resistance to PD-1 and CTLA-4 Blockade and Improves Survival in Pancreatic Carcinoma. Cancer Immunol. Res. 3, 399-41 1 . 10.1 158 / 2326-6066. CIR-14-0215.6. Taube, J.M., Klein, A., Brahmer, J.R., Xu, FL, Pan, X., Kim, J.H., Chen, L., Pardoll, D.M., Topalian, S.L., and Anders, R.A. (2014). Association of PD-1 , PD-1 ligands, and other features of the tumor immune microenvironment with response to anti-PD-1 therapy. Clin. Cancer Res. Off. J. Am. Assoc. Cancer Res. 20, 5064-5074. 10.1 158 / 1078-0432. CCR-13-3271 .7. Chen, D.S., and Mellman, I. (2013). Oncology Meets Immunology: The Cancer-Immunity Cycle. Immunity 39, 1-10. 10.1016 / j.immuni.2013.07.012.8. Sharma, P., and Allison, J.P. (2015). Immune Checkpoint Targeting in Cancer Therapy:Toward Combination Strategies with Curative Potential. Cell 161, 205-214.10.1016 / j.cell.2O15.03.030.9. Pardoll, D.M. (2012). The blockade of immune checkpoints in cancer immunotherapy. Nat. Rev. Cancer 12, 252-264. 10.1038 / nrc3239.10. Robert, C. (2020). A decade of immune-checkpoint inhibitors in cancer therapy. Nat. Commun. 11, 3801. 10.1038 / s41467-020-17670-y.1 1 . Hodi, F.S., O’Day, S.J., McDermott, D.F., Weber, R.W., Sosman, J.A., Haanen, J.B., Gonzalez, R., Robert, C., Schadendorf, D., Hassel, J.C., et al. (2010). Improved survival withipilimumab in patients with metastatic melanoma. N. Engl. J. Med. 363, 71 1-723. 10.1056 / NEJMoa1003466.12. Robert, C., Schachter, J., Long, G.V., Arance, A., Grob, J. J., Mortier, L., Daud, A., Carlino,M.S., McNeil, C., Lotem, M., et al. (2015). Pembrolizumab versus Ipilimumab in Advanced Melanoma.N. Engl. J. Med. 372, 2521 -2532. 10.1056 / NEJMoa1503093.13. Kourie, H.R., and Klastersky, J. (2016). Immune checkpoint inhibitors side effects and management. Immunotherapy 8, 799-807. 10.2217 / imt-2016-0029.14. Postow, M.A., Sidlow, R., and Hellmann, M.D. (2018). Immune-Related Adverse Events Associated with Immune Checkpoint Blockade. N. Engl. J. Med. 378, 158-168. 10.1056 / NEJMra1703481 .15. Schoenfeld, A.J., and Hellmann, M.D. (2020). Acquired Resistance to Immune Checkpoint Inhibitors. Cancer Cell 37, 443-455. 10.1016 / j.ccell.2020.03.017.16. Thommen, D.S., Schreiner, J., Muller, P., Herzig, P., Roller, A., Belousov, A., Umana, P., Pisa, P., Klein, C., Bacac, M., et al. (2015). Progression of Lung Cancer Is Associated with Increased Dysfunction of T Cells Defined by Coexpression of Multiple Inhibitory Receptors. Cancer Immunol. Res. 3, 1344-1355. 10.1 158 / 2326-6066.CIR-15-0097.17. Koyama, S., Akbay, E.A., Li, Y.Y., Herter-Sprie, G.S., Buczkowski, K.A., Richards, W.G., Gandhi, L., Redig, A.J., Rodig, S.J., Asahina, H., et al. (2016). Adaptive resistance to therapeutic PD-1 blockade is associated with upregulation of alternative immune checkpoints. Nat. Commun. 7, 10501 . 10.1038 / ncomms10501 .18. Kawashima, S., Inozume, T., Kawazu, M., Ueno, T., Nagasaki, J., Tanji, E., Honobe, A., Ohnuma, T., Kawamura, T., Umeda, Y., et al. (2021 ). TIGIT / CD155 axis mediates resistance to immunotherapy in patients with melanoma with the inflamed tumor microenvironment. J. Immunother. Cancer 9, e003134. 10.1 136 / jitc-2021 -003134.19. Hung, A.L., Maxwell, R., Theodros, D., Belcaid, Z., Mathios, D., Luksik, A.S., Kim, E., Wu, A., Xia, Y., Garzon-Muvdi, T., et al. (2018). TIGIT and PD-1 dual checkpoint blockade enhances antitumor immunity and survival in GBM. Oncolmmunology 7, e1466769. 10.1080 / 2162402X.2018.1466769.20. Rescue of exhausted CD8 T cells by PD-1 -targeted therapies is CD28-dependent | Science https: / / www.science.Org / doi / full / 10.1 126 / science.aaf0683.21 . Majzner, R.G., Frank, M.J., Mount, C., Tousley, A., Kurtz, D.M., Sworder, B., Murphy, K.A., Manousopoulou, A., Kohler, K., Rotiroti, M.C., et al. (2020). CD58 Aberrations Limit Durable Responses to CD19 CAR in Large B Cell Lymphoma Patients Treated with Axicabtagene Ciloleucel but Can be Overcome through Novel CAR Engineering. Blood 136, 53-54. 10.1 182 / blood-2020- 139605.22. Frangieh, C.J., Melms, J.C., Thakore, P.I., Geiger-Schuller, K.R., Ho, P., Luoma, A.M., Cleary, B., Jerby-Arnon, L., Malu, S., Cuoco, M.S., et al. (2021 ). Multimodal pooled Perturb-CITE- seq screens in patient models define mechanisms of cancer immune evasion. Nat. Genet. 53, 332- 341 . 10.1038 / S41588-021 -00779-1 .23. Ng, S., and A, P. (2012). The biology and therapeutic targeting of the proprotein convertases. Nat. Rev. Drug Discov. 1 1. 10.1038 / nrd3699.24. Frangois, A., Descarpentrie, J., Badiola, I., Siegfried, G., Evrard, S., Pernot, S., and Khatib, A.-M. (2023). Reprogramming immune cells activity by furin-like enzymes as emerging strategy for enhanced immunotherapy in cancer. Br. J. Cancer 128, 1 189-1 195. 10.1038 / s41416-022-02073-1 .25. He, Z., Khatib, A.-M., and Creemers, J.W.M. (2022). The proprotein convertase furin in cancer: more than an oncogene. Oncogene 41, 1252-1262. 10.1038 / s41388-021 -02175-9.26. Rose, M., Duhamel, M., Rodet, F., and Salzet, M. (2021 ). The Role of Proprotein Convertases in the Regulation of the Function of Immune Cells in the Oncoimmune Response. Front. Immunol. 12, 667850. 10.3389 / fimmu.2021 .667850.27. He, Z., Khatib, A.-M., and Creemers, J.W.M. (2020). Loss of Proprotein Convertase Furin in Mammary Gland Impairs prolGFI R and proIR Processing and Suppresses Tumorigenesis in Triple Negative Breast Cancer. Cancers 12, 2686. 10.3390 / cancers12092686.28. Scamuffa, N., Siegfried, G., Bontemps, Y., Ma, L., Basak, A., Cherel, G., Calvo, F., Seidah, N.G., and Khatib, A.-M. (2008). Selective inhibition of proprotein convertases represses the metastatic potential of human colorectal tumor cells. J. Clin. Invest. 118, 352-363. 10.1 172 / JCI32040.29. Thomas, G. (2002). Furin at the cutting edge: from protein traffic to embryogenesis and disease. Nat. Rev. Mol. Cell Biol. 3, 753-766. 10.1038 / nrm934.30. Ojanen, M.J.T., Caro, F.M., Aittomaki, S., Ploquin, M.J., Ortutay, Z., Pekkarinen, M., Kesseli, J., Vahatupa, M., Maatta, J., Nykter, M., et al. (2023). FURIN regulates cytotoxic T- lymphocyte effector function and memory cell transition in mice. Eur. J. Immunol. 53, e2250246. 10.1002 / eji.202250246.31 . Chen, M., Pan, Y., Liu, H., Ning, F., Lu, Q., Duan, Y., Gan, X., Lu, S., Hou, H., Zhang, M., et al. (2023). Ezrin accelerates breast cancer liver metastasis through promoting furin-like convertase-mediated cleavage of Notchl . Cell. Oncol. Dordr. 46, 571-587. 10.1007 / s13402-022- 00761 -x.32. Garten, W., Hallenberger, S., Ortmann, D., Schafer, W., Vey, M., Angliker, H., Shaw, E., and Klenk, H.D. (1994). Processing of viral glycoproteins by the subtilisin-like endoprotease furin and its inhibition by specific peptidylchloroalkylketones. Biochimie 76, 217-225. 10.1016 / 0300- 9084(94)90149-x.33. Klein-Szanto, A.J., and Bassi, D.E. (2017). Proprotein convertase inhibition: Paralyzing the cell’s master switches. Biochem. Pharmacol. 140, 8-15. 10.1016 / j.bcp.2017.04.027.51 . Anderson, E.D., Thomas, L., Hayflick, J.S., and Thomas, G. (1993). Inhibition of HIV-1 gp160-dependent membrane fusion by a furin-directed alpha 1 -antitrypsin variant. J. Biol. Chem. 268, 24887-24891 .52. Bruno, B.J., Miller, G.D., and Lim, C.S. (2013). Basics and recent advances in peptide and protein drug delivery. Ther. Deliv. 4, 1443-1467. 10.4155 / tde.13.104.53. Henrich, S., Cameron, A., Bourenkov, G.P., Kiefersauer, R., Huber, R., Lindberg, I., Bode, W., and Than, M.E. (2003). The crystal structure of the proprotein processing proteinase furin explains its stringent specificity. Nat. Struct. Biol. 10, 520-526. 10.1038 / nsb941 .54. Van Lam van, T., Ivanova, T., Hardes, K., Heindl, M.R., Morty, R.E., Bbttcher- Friebertshauser, E., Lindberg, I., Than, M.E., Dahms, S.O., and Steinmetzer, T. (2019). Design, Synthesis, and Characterization of Macrocyclic Inhibitors of the Proprotein Convertase Furin. ChemMedChem 14, 673-685. 10.1002 / cmdc.201800807.55. Jiao, G.-S., Cregar, L., Wang, J., Millis, S.Z., Tang, C., O’Malley, S., Johnson, A.T., Sareth, S., Larson, J., and Thomas, G. (2006). Synthetic small molecule furin inhibitors derived from 2,5- dideoxystreptamine. Proc. Natl. Acad. Sci. U. S. A. 103, 19707-19712. 10.1073 / pnas.0606555104

Claims

Claims

1. Compound of formula (I)wherein Ri , R2, R3, R4, R5, Re, R7, Rs, R9, R10, R11 are independently selected from hydrogen, halogen, amine, Ci-s alkyl, C1-8 alkoxy, -NO2, -CN, optionally substituted phenyl, and optionally substituted heterocycle, for use in a method of treatment of cancer, said method comprising administering to a subject in need thereof an effective amount of compound of formula (I) or of a pharmaceutically acceptable salt thereof.

2. Compound for use in a method according to claim 1 , wherein said compound is selected from compounds of formula (la) or (lb)wherein R1 , R3, R4, R5, Re, R7, Rs, R9, R10, R11 , R12, R13, R14, R15, R16, R17, Ris, R19, R20 are independently selected from hydrogen, halogen, amine, Ci-s alkyl, Ci-s alkoxy, -NO2, -CN, optionally substituted phenyl, and optionally substituted heterocycle.

3. Compound for use in a method according to claim 1 or 2, wherein said compound is selected from compounds of formula (Ia1 ), (Ia2) or (Ia3)selected from hydrogen, halogen, amine, C1-8 alkyl, C1-8 alkoxy, -NO2, -CN, optionally substituted phenyl, and optionally substituted heterocycle.

4. Compound for use in a method according to anyone of claims 1 to 3, wherein said compound is selected from

5. Compound for use in a method according to any one of claims 1 to 4 in combination with at least one other anticancer therapy.

6. Compound for use in a method according to claim 5, wherein the anticancer therapy is a chemotherapy, targeted therapy, immunotherapy or radiation therapy.

7. Compound for use in a method according to any one of claims 1 to 6, wherein the cancer is selected from colon cancer, liver cancer, pancreatic cancer, leukemia, prostate cancer and breast cancer.

8. Compound for use in a method according to any one of claims 1 to 7, wherein the subject is human.

9. Pharmaceutical composition comprising a compound of formula (I)whereinRi , R2, R3, R4, R5, Re, R7, Rs, R9, R10, R11 are independently selected from hydrogen, halogen, amine, C1-8 alkyl, C1-8 alkoxy, -NO2, -CN, optionally substituted phenyl, and optionally substitutedheterocycle, or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient, for use in a method of treatment of cancer.

10. Pharmaceutical composition for use according to claim 9 wherein said compound is selected from compounds of formula (la) or (lb)whereinRi , R3, R4, R5, Re, R7, Rs, R9, R10, R11 , R12, R13, R14, R15, R16, R17, Ris, R19, R20 are independently selected from hydrogen, halogen, amine, C1-8 alkyl, C1-8 alkoxy, -NO2, -CN, optionally substituted phenyl, and optionally substituted heterocycle.

11. Pharmaceutical composition for use according to claim 9 or 10 wherein said compound is selected from compounds of formula (Ia1 ), (Ia2) or (Ia3)whereinRi , R3, R4, R5, Re, R7, Rs, R9, R10, R11 , R12, R13, R14, R15, R16, R17, Ris, R19, R20 are independently selected from hydrogen, halogen, amine, C1-8 alkyl, C1-8 alkoxy, -NO2, -CN, optionally substituted phenyl, and optionally substituted heterocycle.

12. Pharmaceutical composition for use according to anyone of claims 9 to 1 1 wherein said compound is selected from

13. Compound for use in a method according to any one of claims 1 to 9 or pharmaceutical composition for use according to any one of claims 9 to 12, wherein said compound or composition is administered orally, enterally, intravenously, or intramuscularly.

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