Disulfonate stilbenes for use in the treatment of proliferative disorders - Patents.com

Abstract

The present invention relates to compounds of the general formula: 0A and R 0B is independently selected from hydrogen and a pharmaceutically acceptable cation; R A and R B are the same and are selected from amide, carbamate, sulfonamide, azide, cyano, and halide. The present invention also relates to pharmaceutical compositions comprising the compounds of the present invention. According to one embodiment, the composition further comprises another active ingredient, in particular an anti-neoplastic agent. The present invention also relates to compounds or compositions of the present invention for use as a medicament, in particular for use in the treatment of proliferative diseases such as cancer. [Case 1] TIFF2022514195000027.tif46162

Description

[Technical field]

[0001] The present invention relates to disulfonate stilbene compounds, in particular disulfonate stilbenes, useful as inhibitors of the RAD51 protein for use in the treatment of proliferative diseases. [Background technology]

[0002] Proliferative disease therapy such as radiotherapy or chemotherapy often faces the resistance induced and / or inherent resistance from cancer cells.DNA repair pathway can repair the DNA of cancer cells that is damaged by this treatment, which reduces the efficacy of the treatment.Therefore, inhibiting DNA repair pathway is an interesting option for sensitizing tumors to anti-cancer treatment.

[0003] Double-strand breaks (DSBs) are the most damaging nuclear alterations in DNA and can be repaired by a biological process called homologous recombination (HR). RAD51 protein is a key component of the HR repair pathway. The first step of the recombinase activity of RAD51 protein is the formation of nucleofilaments of RAD51 protein that catalyze the search for homology and strand exchange between the damaged DNA sequence and the homologous sequence, thus ensuring accurate repair of DSBs. Thus, dysregulation of RAD51 protein can lead to carcinogenesis and promote cancer resistance. Overexpression of RAD51 protein leads to genomic instability in cells at the origin of some cancers and also provides resistance by promoting the repair of DSBs induced by some anticancer treatments. In particular, it has been shown that patients who are being treated for cancer and express higher levels of RAD51 protein have shorter survival, and that reducing the amount of RAD51 protein by antisense or ribozyme treatment improves the efficacy of cancer treatment by radiotherapy. Thus, the RAD51 protein is a suitable biological target for regulating HR, thus resulting in better efficacy of antiproliferative treatments.

[0004] Attention has been focused on identifying small molecules that exhibit anti-recombinase activity, and several RAD51 protein inhibitory small molecules have been found. These can directly interfere with certain steps of RAD51 protein recombinase activity, thereby preventing or limiting the HR repair pathway. Examples of RAD51 protein small molecule inhibitors in the field include: [ka]

[0005] In particular, it has been shown that BPQ impairs the formation of RAD51 filaments by targeting protein-DNA interactions (Huang, F. et al., "Inhibition of Homologous Recombination in Human Cells by Targeting RAD51 Recombinase", Journal of Medicinal Chemistry, April 2012, Vol. 55, pp. 3011-3020), and that sensitization of cancer cells treated with doxorubicin can be observed (Alagpulinsa, DA et al., "A small-molecule inhibitor of RAD51 reduces homologous recombination and sensitizes multiple myeloma cells to doxorubicin", Frontiers in Oncology, October 2014, Vol. 4, p. 289).

[0006] In particular, it has been shown that DIDS can inhibit the recombinase activity of RAD51 during HR (Ishida, T., et al., "DIDS, a chemical compound that inhibits RAD51-mediated homologous pairing and strand exchange.", Nucleic Acids Research, 2009, Vol. 37, No. 10, pp. 3367-3376). The inhibitory activity is likely due to the direct interaction of DIDS with RAD51 protein, but the mechanism of action of DIDS has not been elucidated and the biological targets involved remain to be determined.

[0007] However, small molecule inhibitors of the RAD51 protein, such as DIDS or BPQ, have limitations that impede their medical use against cancer, e.g., regarding therapeutic efficiency (e.g., with respect to sensitizing effect), the presence or potency of side effects, selectivity over other biological receptors, and / or the chemical stability of the molecule.

[0008] In particular, DIDS as an inhibitor is not specific to RAD proteins because it also inhibits membrane transporters (Wilson, MC, et al. "Studies on the DIDS-binding site of monocarboxylate transporter 1 suggest a homology model of the open conformation and a plausible translocation cycle," Journal of Biological Chemistry, 2009, Vol. 284, No. 30, pp. 20011 20021) and ion channels (Matulef, K., et al. "Discovery of potent CLC chloride channel inhibitors," ACS Chemical Biology, 2008, Vol. 3, No. 7, pp. 419 428). Therefore, DIDS may induce adverse side effects such as cytotoxicity.

[0009] Furthermore, the isothiocyanate functional group of DIDS is known to be sensitive to hydrolysis (Jakobsen, P. and Horobin, RW, "Preparation and Characterization of 4-Acetamido-4'-Isothiocyanostilbene-2,2'-Disulfonic Acid (Sits) And Related Stilbene Disulfonates" Stain Technology, 1989, Vol. 64, No. 6, pp. 301-313), and as a result, DIDS oligomers with potential chloride channel or transporter inhibitory activity can be formed (see, e.g., International Patent Publication No. WO 2010 / 148177).

[0010] The applicant has carried out extensive research to envision and synthesize novel RAD51 protein inhibitors that can overcome the limitations of known compounds.Unexpectedly, during the course of this research, it has been found that some disulfonatostilbene compounds are potent inhibitors of RAD51 protein and thus may be useful in the treatment of proliferative diseases.

[0011] The disulfonatostilbenes of the present invention are potent RAD51 protein inhibitors, with efficacy comparable to or superior to that of RAD51 proteins in the art, and in particular, they efficiently sensitize proliferating cells in the presence of anti-cancer drugs.

[0012] The disulfonate stilbenes of the present invention induce fewer adverse side effects than prior art RAD51 inhibitors, in particular they are less cytotoxic.

[0013] The disulfonatostilbenes of the present invention may also have improved chemical stability compared to molecules in the art. Summary of the Invention

[0014] The present invention relates to a compound of formula (I): [ka] (In the formula, R 0A and R 0B is independently selected from hydrogen, lithium, sodium, and potassium; R A and R B are identical, -NH-COR, -NH-COOR, -NH-SO 2 R is selected from azide, cyano, and halide; where each R is independently: (C 1 -C 6 )-alkyl, (C 3 -C 6 )-cycloalkyl, (C 6 -C 10 ) aryl, and (C 6 -C 10 )-heteroaryl groups; The groups are independently selected from the group 1 -C 6 )-alkyl, (C 3 -C 6 )-cycloalkyl, (C 1 -C 6 )-alkoxy, (C 1 -C 6 )-alkyl-S-, and nitro) or a pharma- ceutically acceptable salt thereof.

[0015] According to one embodiment, each R is independently selected from the group consisting of: 1 -C 4 )-alkyl, phenyl, pyridinyl, or diazinyl groups; the phenyl, pyridinyl, or diazinyl groups are independently selected from (C 1 -C 3 )-alkyl, (C 1 -C 3)-alkoxy, and nitro, preferably methyl, methoxy, and nitro. According to one embodiment, R A and R B -NH-COR, -NH-COOR, and -NH-SO 2 R. According to one embodiment, R A and R B is selected from azide, cyano, and a halide. According to one embodiment, the halide is iodine. According to one embodiment, R 0A and R 0B are both sodium or R 0A and R 0B Both are potassium.

[0016] According to one embodiment, the compound is Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-acetamidobenzenesulfonate); Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-propionamidobenzenesulfonate); Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-benzamidobenzenesulfonate); Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-(4-methoxybenzamido)benzenesulfonate); Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-(4-nitrobenzamido)benzenesulfonate); Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-((ethoxycarbonyl)amino)benzenesulfonate); Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-((isobutoxycarbonyl)amino)benzenesulfonate); Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-((phenoxycarbonyl)amino)benzenesulfonate); Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-(phenylsulfonamido)benzenesulfonate); Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-((4-methylphenyl)sulfonamido)benzenesulfonate); Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-((4-nitrophenyl)sulfonamido)benzenesulfonate); Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-azidobenzenesulfonate); Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-cyanobenzenesulfonate); and Potassium (E)-6,6'-(ethene-1,2-diyl)bis(3-iodobenzenesulfonate) is selected from.

[0017] According to one embodiment, the proliferative disease is cancer, preferably breast cancer, glioblastoma, or multiple myeloma.

[0018] The present invention also relates to a composition comprising a compound of formula (I) and a pharma- ceutically acceptable excipient for use according to the present invention.

[0019] According to one embodiment, the composition further comprises at least another active ingredient, hi one embodiment, the at least another active ingredient is an antineoplastic active ingredient.

[0020] The present invention also relates to a compound of formula (II): [ka] (In the formula, R 0A and R 0B is independently selected from hydrogen, lithium, sodium, and potassium; Each R is independently: (C 6 -C 10 )-aryl and (C 6 -C10 )-heteroaryl groups) or a pharma- ceutically acceptable salt thereof.

[0021] According to one embodiment, each R is independently selected from phenyl, pyridinyl, and diazinyl; preferably phenyl. According to one embodiment, R 0A and R 0B are identical; preferably R 0A and R 0B is sodium.

[0022] The present invention also relates to a process for preparing a compound of formula (II) according to the invention, comprising the step of reacting a halo-formate with (E) 6,6'-(ethene-1,2-diyl)bis(3-aminobenzenesulfonate). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0023] definition In the present invention, the following terms have the following meanings.

[0024] "Active ingredient" and "active pharmaceutical ingredient" and "therapeutic agent" refer to a compound for use in therapy and related to health. In particular, an active ingredient may be adapted to treat or prevent a disease, preferably a proliferative disease. An active ingredient may also be adapted to improve the therapeutic activity of another active ingredient to treat or prevent a disease, preferably a proliferative disease.

[0025] "Alkene" or "alkenyl" refers to any straight or branched hydrocarbon chain having from 2 to 12 carbon atoms, preferably from 2 to 8 carbon atoms, more preferably from 1 to 6 carbon atoms, and even more preferably from 1 to 4 carbon atoms; and having at least one double bond. Examples of alkenyl groups are ethenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl and its isomers, 2-hexenyl and its isomers, or 2,4-pentadienyl.

[0026] "Alkoxy" refers to a radical of the formula -O-alkyl.

[0027] "Alkyl" refers to any straight or branched saturated hydrocarbon chain having 1 to 12 carbon atoms, preferably 1 to 8 carbon atoms, more preferably 1 to 6 carbon atoms, and even more preferably 1 to 4 carbon atoms. Examples of alkyl groups are methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, pentyl and its isomers (e.g., n-pentyl or i-pentyl), or hexyl and its isomers (e.g., n-hexyl or i-hexyl).

[0028] "Alkyne" or "alkynyl" refers to any straight or branched hydrocarbon chain having from 2 to 12 carbon atoms, preferably from 2 to 8 carbon atoms, more preferably from 1 to 6 carbon atoms, and even more preferably from 1 to 4 carbon atoms; and having at least one triple bond. Examples of alkynyl groups are ethynyl, 2-propynyl, 2-butynyl, 3-butynyl, 2-pentynyl and its isomers, or 2-hexynyl and its isomers.

[0029] "Amido" refers to a group containing an amide moiety (-NH-CO-).

[0030] "Aryl" refers to a polyunsaturated aromatic hydrocarbon group having 5 to 16 carbon atoms, preferably 6 to 12 carbon atoms, more preferably 6 to 10 carbon atoms; having a single ring (i.e., phenyl) or multiple aromatic rings either fused together (e.g., naphthyl) or covalently linked, in which at least one ring is aromatic. The aromatic ring may optionally contain one to two additional rings (either cycloalkyl, heterocyclyl, or heteroaryl) fused thereto. Examples of aryl are phenyl, biphenylyl, biphenylenyl, 5- or 6-tetralinyl, naphthalen-1- or -2-yl, 4-, 5-, 6, or 7-indenyl, 1-, 2-, 3-, 4-, or 5-acenaphtylenyl, 3-, 4-, or 5-acenaphtenyl, 1- or 2-pentalenyl, 4- or 5-indanyl, 5-, 6-, 7-, or 8-tetrahydronaphthyl, 1,2,3,4-tetrahydronaphthyl, 1,4-dihydronaphthyl, 1-, 2-, 3-, 4-, or 5-pyrenyl.

[0031] "Azide" is a compound of the formula -N 3 represents a group.

[0032] "BPQ" stands for (E)-3-benzyl-2-(2-(pyridin-3-yl)vinyl)quinazolin-4(3H)-one.

[0033] "Carbamate" refers to a group that contains a carbamate moiety (-NH-COO-).

[0034] "Ctrl" stands for "control."

[0035] Cyano represents a group of the formula -CN.

[0036] "Cycloalkyl" refers to any cyclic or polycyclic alkyl group containing at least three carbon atoms, which is linear or branched. Examples of cycloalkyl groups are cyclopropyl, cyclopentyl, or cyclohexyl.

[0037] "Cycloalkenyl" refers to any cyclic or polycyclic alkenyl group that is linear or branched and contains at least 5 carbon atoms.

[0038] "DIDS" stands for 4,4'-diisothiocyano-2,2'-stilbene-disulfonic acid.

[0039] "Disease" includes any disease, disorder, or condition, preferably a proliferative disease, disorder, or condition. The goal of treatment is to prevent or delay disease in a subject in need thereof.

[0040] "DNA" stands for deoxyribonucleic acid.

[0041] "Halide" refers to a fluorine (F), chlorine (Cl), bromine (Br), or iodine (I) atom.

[0042] "Heteroaryl" refers to an aromatic ring or ring system having 5 to 16 carbon atoms, preferably 6 to 12 carbon atoms, more preferably 6 to 10 carbon atoms; having one or two rings fused or covalently bonded together, in which at least one ring is aromatic; in which one or more of the rings one or more carbon atoms are replaced by the elements oxygen, nitrogen, and / or sulfur, the nitrogen and sulfur heteroatoms may be optionally oxidized, and the nitrogen heteroatom may be optionally quaternized. Such rings may be fused to an aryl, cycloalkyl, heteroaryl, or heterocyclyl ring. Examples of heteroaryl are furanyl, thiophenyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, oxatriazolyl, thiatriazolyl, pyridinyl, pyrimidyl, pyrazinyl, pyridazinyl, oxazinyl, dioxinyl, thiazinyl, triazinyl, imidazo[2,1-b][1,3]-thiazolyl or thieno[3,2-b]furanyl.

[0043] "Heteroalkyl" refers to any alkyl group containing at least three carbon atoms and interrupted by at least one heteroatom selected from oxygen, nitrogen, and sulfur. The nitrogen and sulfur heteroatoms may be optionally oxidized, and the nitrogen heteroatom may be optionally quaternized.

[0044] "Heterocycloalkyl" refers to a non-aromatic cyclic or polycyclic alkyl group containing at least three carbon atoms, linear or branched, and interrupted by at least one heteroatom selected from oxygen, nitrogen, and sulfur. The nitrogen and sulfur heteroatoms may be optionally oxidized, and the nitrogen heteroatom may be optionally quaternized.

[0045] "Nitro" is a group of the formula -NO 2 represents a group.

[0046] "Pharmaceutical composition" refers to a composition comprising an active ingredient in association with a pharma- ceutically acceptable excipient. The pharmaceutical composition is for therapeutic use and is health-related. In particular, the pharmaceutical composition may be adapted to treat or prevent a disease, preferably a proliferative disease.

[0047] "Pharmaceutically acceptable" means that the components of a pharmaceutical composition are compatible with each other and not harmful when administered to a subject.

[0048] A "pharmaceutically acceptable excipient" refers to an excipient or vehicle that does not produce adverse, allergic, or other undesirable reactions when administered to a subject. This includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc. For human administration, preparations must meet sterility, pyrogenicity, general safety, and purity standards as required, for example, by the United States Food and Drug Administration (FDA) or the European Medicines Agency (EMA).

[0049] "RAD51," "RAD51A," or "RECA" refer to the RAD51 gene.

[0050] "RAD51 protein" is an abbreviation for "DNA repair protein RAD51 protein homolog 1" or "RAD recombinase" and refers to the protein encoded by RAD51.

[0051] "RAD51 protein nucleofilaments" represent presynaptic filament assemblies involved in key steps of the RAD51 protein-dependent homologous recombination (HR) process (Velic, D. et al., "DNA Damage Signalling and Repair Inhibitors", Biomolecules, November 2015, Vol. 5, No. 4, pp. 3204-3059).

[0052] "Subject" refers to a warm-blooded animal, preferably a mammal, more preferably a human. Preferably, the subject is a patient, i.e. a subject waiting for medical treatment, receiving medical treatment, or currently / in the future being / will be a subject of medical treatment. Preferably, the subject has a proliferative disease.

[0053] "Sulfonamide" refers to the sulfonamide moiety (-NH-SO 2 -) is represented as a group.

[0054] "Treating" or "treatment" or "alleviation" refers to both therapeutic treatment and prophylactic or preventative measures, the purpose of which is to prevent or delay (attenuate) the disease of interest in a subject in need thereof. Those in need of treatment include those who already have the disease, disorder, or condition, and those who are prone to have the disorder, or those in whom the disorder is to be prevented. A subject is successfully "treated" for said disease, disorder, or condition if, after administration of a therapeutic amount of a substance or composition according to the invention, the subject shows an observable and / or measurable reduction or absence of one or more of the following: a reduction in the number of pathogenic cells; a reduction in the percentage of total cells that are pathogenic; an alleviation to some extent of one or more of the symptoms associated with the particular disease, disorder, or condition; a reduction in morbidity and mortality; and / or an improvement in quality of life issues. The above parameters for assessing the success of treatment and the improvement of the disease, disorder, or condition are easily measurable by routine procedures known to physicians. Preferably, the disease to be treated is a proliferative disease.

[0055] When a chemical substituent is a combination of chemical groups, the point of attachment of the substituent to the molecule is through the last chemical group listed. For example, an "arylalkyl" substituent is attached to the remainder of the molecule through the alkyl moiety, which may be represented as follows: "aryl-alkyl-".

[0056] Detailed Description compound The present invention relates to a compound of formula (I-0): [ka] (In the formula, R 0A and R 0B is independently selected from hydrogen; a pharma- ceutically acceptable cation; and a sulfonate functional protecting group or precursor; R A and R Bis independently selected from amide, carbamate, sulfonamide, azide, cyano, and halide. Regarding.

[0057] According to one embodiment, R A and R B are identical. In one embodiment, R A and R B are the same amide. In one embodiment, R A and R B are the same carbamide. In one embodiment, R A and R B are the same sulfonamide. In one embodiment, R A and R B is azide. In one embodiment, R A and R B is cyano. In one embodiment, R A and R B is a halide.

[0058] According to one embodiment, R A and R B is independently selected from amide, carbamate, and sulfonamide. According to one embodiment, the amide is -NH-COR, the carbamate is NH-COOR, and / or the sulfonamide is -NH-SO 2 R; each R is independently selected from alkyl, arylalkyl, heteroarylalkyl, cycloalkyl, arylcycloalkyl, heteroarylcycloalkyl, alkenyl, arylalkenyl, heteroarylalkenyl, cycloalkenyl, arylcycloalkenyl, heteroarylcycloalkenyl, alkynyl, arylalkynyl, heteroarylalkynyl, aryl, alkylaryl, cycloalkylaryl, alkenylaryl, cycloalkenylaryl, alkynylaryl, heteroaryl, alkylheteroaryl, cycloalkylheteroaryl, alkenylheteroaryl, cycloalkenylheteroaryl, and alkynylheteroaryl groups; The groups or substituents are optionally interrupted by at least one substituent selected from oxygen (O), nitrogen (N), and sulfur (S); The group can be an alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl, amino (-NH 2 ), azide (-N 3 ), cyano (-CN), nitro (-NO 2 ), oxo (=O), halide, hydroxyl (-OH), formyl (-C(O)H), carboxyl (-COOH), amide (-C(O)-NH 2 ), and optionally substituted with at least one substituent selected from thio (=S) and sulfhydryl (-SH); A nitrogen or sulfur atom substituted or contained in a group or substituent is optionally oxidized, and a nitrogen atom substituted or contained in a group or substituent is optionally quaternized.

[0059] In one embodiment, R is selected from an alkyl group, a cycloalkyl group, a heteroalkyl group, a heterocycloalkyl group, an aryl group, and a heteroaryl group, which are optionally interrupted, substituted, oxidized, and / or quaternized as described hereinabove. In one specific embodiment, R is selected from an alkyl group, a cycloalkyl group, an aryl group, and a heteroaryl group, which are optionally interrupted, substituted, oxidized, and / or quaternized as described hereinabove. In one further specific embodiment, R is selected from (C 1 -C 6 )-Alkyl(C 3 -C 6 )-cycloalkyl group, (C 6 -C 10 )-aryl groups, and (C 6 -C 10)-heteroaryl groups, which are optionally interrupted, substituted, oxidized, and / or quaternized as described herein above. In a further specific embodiment, R is selected from alkyl groups and aryl groups, which are optionally interrupted, substituted, oxidized, and / or quaternized as described herein above. In a further specific embodiment, R is selected from alkyl groups and cycloalkyl groups, which are optionally interrupted, substituted, oxidized, and / or quaternized as described herein above. In a further specific embodiment, R is selected from aryl groups and heteroaryl groups, which are optionally interrupted, substituted, oxidized, and / or quaternized as described herein above.

[0060] In one embodiment, R is as defined hereinabove, which is optionally interrupted, oxidized, or quaternized as described hereinabove; and which is optionally substituted with at least one substituent selected from alkyl, cycloalkyl, alkoxy, alkyl-S-, and nitro. In one specific embodiment, R is as defined hereinabove, which is optionally interrupted, oxidized, or quaternized as described hereinabove, and which is optionally selected from (C 1 -C 6 )-alkyl, (C 3 -C 6 )-cycloalkyl, (C 1 -C 6 )-alkoxy, (C 1 -C 6 )-alkyl-S, and nitro, preferably (C 1 -C 3 )-alkyl, (C 1 -C 3 )-alkoxy, and nitro; more preferably (C 1 -C 2 )-alkyl, (C 1 -C 2)-alkoxy, and nitro, and even more preferably substituted with at least one substituent selected from methyl, methoxy, and nitro. In one specific embodiment, R is as defined hereinabove, which group is optionally interrupted, oxidized, or quaternized as described hereinabove; and optionally substituted with at least one substituent selected from alkyl, alkoxy, and nitro. In one embodiment, R is as defined hereinabove, which group is optionally interrupted, oxidized, or quaternized as described hereinabove; and optionally substituted with at least one substituent selected from (C 1 -C 3 )-alkyl, (C 1 -C 3 )-alkoxy, and nitro; preferably (C 1 -C 2 )-alkyl, (C 1 -C 2 )-alkoxy, and nitro; more preferably substituted with at least one substituent selected from methyl, methoxy, and nitro.

[0061] In a more specific embodiment, R is selected from an alkyl group, a cycloalkyl group, an aryl group, and a heteroaryl group; this group is optionally substituted with at least one substituent selected from alkyl, cycloalkyl, alkoxy, alkyl-S-, and nitro. In a further specific embodiment, R is selected from an alkyl group and an aryl group; the aryl group is optionally substituted with at least one substituent selected from alkyl, alkoxy, and nitro. For example, R as an alkyl group can be methyl, ethyl, propyl, and butyl. For example, R as an aryl group can be phenyl that does not contain any substituent or phenyl that is substituted with only one group selected from methyl, methoxy, and nitro.

[0062] In one more specific embodiment, R is selected from the group consisting of (C 1 -C 4)-alkyl, for example methyl, ethyl, propyl (eg n-propyl and isopropyl), and butyl (eg n-butyl, isobutyl and tert-butyl).

[0063] In another more specific embodiment, R is phenyl, which is optionally interrupted, substituted, oxidized, and / or quaternized as described hereinabove.

[0064] In another more specific embodiment, R is pyridinyl or diazinyl, which is optionally interrupted, substituted, oxidized, and / or quaternized as described herein above. The pyridinyl or diazinyl group may be attached to an amide, carbamate, or sulfonamide functional group through either a carbon atom (e.g., 2-pyridinyl, 3-pyridinyl, 4-pyridinyl, 5-pyridinyl, or 6-pyridinyl) or nitrogen atom (e.g., 1-pyridinyl) of the pyridinyl or diazinyl; preferably through a carbon atom of the pyridinyl or diazinyl.

[0065] In one embodiment, R is substituted with one or two substituents as described hereinabove. In one specific embodiment, R is substituted with only one substituent as described hereinabove.

[0066] According to one embodiment, R A and R B is independently selected from azide, cyano, and a halide. In one embodiment, the halide is an iodine (I) atom.

[0067] According to one embodiment, R A and R B At least one of R is not an amide, in particular is not -NH-COR, where R is as defined herein above. A and R B and R are not both amides. A and R BAt least one of the groups is not a sulfonamide, particularly -NH-SO 2 R, where R is as defined herein above. In one embodiment, R A and R B are not both sulfonamides. According to one embodiment, at least one of the R groups is not alkyl. In one embodiment, the R group is not alkyl. According to one embodiment, at least one of the R groups is not cycloalkyl. In one embodiment, the R group is not cycloalkyl. According to one embodiment, at least one of the R groups is a sulfonate functional group (-SO 2 In one embodiment, the R group is not substituted with a sulfonate functional group.

[0068] According to one embodiment, R 0A and R 0B are identical. In one embodiment, R 0A and R 0B is hydrogen. In one embodiment, R 0A and R 0B are the same pharma- ceutically acceptable cations.

[0069] According to one embodiment, R 0A and / or R 0B is a pharma- ceutically acceptable alkaline cation. In one embodiment, the pharma- ceutically acceptable alkaline cation is lithium (Li + ), Sodium (Na + ), and potassium (K + ). Preferably, the pharma- ceutically acceptable alkaline cation is selected from sodium (Na + ) or potassium (K + ), more preferably sodium (Na + ).

[0070] According to one embodiment, R 0A and / or R 0Bis an alkyl group as described herein above, such as ethyl, isopentyl, methyl, ethyl, isopropyl, or neopentyl. In this embodiment, the compounds of formula (I-0) are useful, for example, as synthetic precursors.

[0071] According to one embodiment, R 0A and / or R 0B is an aryl group as described herein above, such as phenyl or para-nitrophenyl. In one embodiment, the alkyl group is selected from ethyl. In this embodiment, the compounds of formula (I-0) are useful, for example, as prodrugs.

[0072] According to one embodiment, R 0A and / or R 0B is a group that contains a functional group that can be converted by an enzyme system (e.g., nitroreductase or esterase) to a free sulfonate functional group, such as those disclosed in International Patent Publication No. WO 2011 / 017800 (Kong, X. et al.). In this embodiment, the compounds of formula (I-0) are useful, for example, as prodrugs.

[0073] According to one embodiment, the compound according to the invention is a compound of formula (Ia): [ka] (In the formula, R 0A and / or R 0B is as described herein above)

[0074] In one embodiment, the compound of the invention is a compound of formula (Iai). [ka] (In the formula, R A is as described herein above)

[0075] In one embodiment, the compound of the invention is a compound of formula (Ia-ii). [ka] (In the formula, R A is as described herein above)

[0076] According to one embodiment, the compound according to the invention is a compound of formula (Ib). [ka] (In the formula, R A and R B is as described herein above)

[0077] According to one embodiment, the compound according to the invention is a compound of formula (Ic): [ka] (In the formula, R A and R B is as described herein above)

[0078] According to one embodiment, the compound according to the invention is a compound of formula (II-0): [ka] (In the formula, R A and R B and each R is as described herein above.

[0079] According to one embodiment, the compound according to the invention is a compound of formula (III-0): [ka] (In the formula, R A and R B is as described herein above)

[0080] In one specific embodiment, the compound according to the invention is selected from the compounds in Table 1 below. [Table 1] TIFF0007611824000013.tif241164TIFF0007611824000014.tif182165

[0081] The compounds in Table 1 were named using ChemDraw® Professional 15.0 (PerkinElmer).

[0082] According to one embodiment, R A and / or R B is not a thiocyanate. According to one embodiment, R A and / or R B is not nitro.

[0083] In one embodiment, R is not methyl. According to one embodiment, R is not ethyl.

[0084] Reference to a compound of the present invention includes all enantiomers, solvates (e.g., hydrates), polymorphs (e.g., crystalline forms), multicomponent complexes, and pharma- ceutically acceptable salts thereof. Reference to a compound of the present invention further includes all prodrugs thereof.

[0085] Reference to the compounds of the present invention includes all pharma- ceutically acceptable salts thereof. Pharmaceutically acceptable salts include the acid addition and base addition salts thereof. Suitable acid addition salts are formed from acids which form non-toxic salts. Examples include acetate, adipate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, camsylate, citrate, cyclamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hybenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, phosphate ... The salts include gonate, maleate, malonate, mesylate, methylsulfate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogen, phosphate / dihydrogen, phosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate, and xinafoate. Suitable bases are formed from bases that form non-toxic salts. Examples include aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine, 2-(diethylamino)ethanol, ethanolamine, morpholine, 4-(2-hydroxyethyl)morpholine, and zinc salts. Hemisalts of acids and bases can also be formed, such as hemisulfate and hemicalcium salts. When the compound contains an acidic and a basic group, it may also form an internal salt. When the compound contains a hydrogen-donating heteroatom (e.g., NH), its pharma- ceutically acceptable salts also include salts and / or isomers formed by the transfer of said hydrogen atom to a basic group or atom within the molecule. These salts may be prepared by standard procedures, for example, by reacting the free acid with an appropriate organic or inorganic base.

[0086] Compounds that do not have sulfonate functionality and are not part of this invention are presented in Table 2 below. [Table 2]

[0087] The compounds in Table 2 were named using ChemDraw Professional 15.0 (PerkinElmer). The compounds in Table 2 are used for comparison in the "Examples" section below. They are not part of the present invention.

[0088] synthesis The compounds according to the invention described hereinabove may be prepared by any synthetic method known in the art.

[0089] According to one embodiment, the compound is sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-aminobenzenesulfonate) (DADS): [ka] It is manufactured starting from

[0090] In one embodiment, the compound is prepared by the reaction of an acid halide or anhydrous halide, such as an acid chloride or anhydrous chloride, with DADS. In one embodiment, the compound is prepared by the reaction of a halo-formate, such as a chloroformate, with DADS. In one embodiment, the compound is prepared by the reaction of a sulfonyl halide, such as a sulfonyl chloride, with DADS. In one specific embodiment, the reaction is carried out in a basic medium, for example with Na 2 CO 3 or N,N-diisopropylethylamine (DIEA). In one specific embodiment, the reaction is carried out in the presence of a solvent such as a water / dioxane mixture, dimethylsulfoxide (DMF) or dichloromethane (CH 2 Cl 2) or the like. In one specific embodiment, a catalyst such as tetrabutylammonium chloride may be present. In one specific embodiment, the reaction is carried out at room temperature or at 80° C.

[0091] These embodiments are illustrated in Scheme 1 below. [ka]

[0092] In one embodiment, the compounds are prepared starting from DADS by the displacement of the aromatic amino group via the preparation of a diazonium double salt followed by substitution with a nucleophile (Sandmeyer reaction), the nucleophile being, for example, N - 3 , C.N. - , or I - It is.

[0093] In one specific embodiment, the preparation of the diazonium double salt is carried out by using Na 2 NO 2 and acetic acid (AcOH). In one specific embodiment, the nucleophile is a sodium or potassium salt. In one specific embodiment, the displacement step is carried out in the presence of a catalyst, such as a copper (I) or (II) catalyst, such as copper iodide (CuI). In one specific embodiment, the first and / or second reaction step is carried out in a solvent, such as water. In one specific embodiment, the first and / or second reaction step is carried out at room temperature.

[0094] composition The present invention also relates to compositions comprising the compounds according to the invention as described herein above.

[0095] According to one embodiment, the composition further comprises at least another active ingredient, i.e., a compound according to the invention and another different active ingredient. In one embodiment, the active ingredient is an antineoplastic agent, i.e., a compound effective in treating proliferative diseases. In one specific embodiment, the active ingredient is selected from alkylating agents, antimetabolites, microtubule inhibitors, topoisomerase inhibitors, and cytotoxic antibiotics. Examples of antineoplastic agents are listed in Actualite Pharmaceutiques, October 1992 (No. 302, pp. 41-43), which is incorporated herein by reference. In one further specific embodiment, the active ingredient is selected from alkylating agents selected from nitrogen mustards, nitrosoureas, tetrazines, aziridines, cisplatin, and cisplatin derivatives (e.g., carboplatin and oxaliplatin), and non-classical alkylating agents such as procarbazine and hexamethylmelamine. Examples of antineoplastic agents include cisplatin or its derivatives; inhibitors of topoisomerase, such as etoposide or topotecan; or tyrosine kinase receptors, such as erlotinib or imatinib.

[0096] According to one embodiment, the composition further comprises a pharma- ceutically acceptable excipient. In one embodiment, the composition is a pharmaceutical composition. In one embodiment, the composition is a drug.

[0097] kit The present invention also relates to a kit comprising a compound or composition according to the invention as described herein above.

[0098] According to one embodiment, the kit comprises at least one pharmaceutical composition according to the invention and at least another pharmaceutical composition comprising another active ingredient.

[0099] According to one embodiment, the kit comprises a plurality of pharmaceutical compositions according to the invention, eg, a plurality of single doses for monthly, weekly or daily administration.

[0100] Uses and Methods

[0101] The present invention also relates to the use of a compound or composition according to the invention as described hereinabove for inhibiting DNA repair, e.g. for inhibiting repair of a double strand break.The present invention also relates to a method for inhibiting DNA repair (e.g. for example repair of a double strand break) in a subject in need thereof, comprising administering to the subject a compound or composition according to the invention as described hereinabove.

[0102] According to one embodiment, the inhibition of double strand break repair comprises the inhibition of RAD51 protein. The inhibition of RAD51 protein includes any reduction in the presence of RAD51 protein, for example, preventing RAD51 protein synthesis, and / or preventing the folding of RAD51 protein. The inhibition of RAD51 protein also includes any reduction in the biological activity of RAD51 protein, for example, preventing the formation of RAD51 protein nucleofilaments, the binding of RAD51 protein to DNA, and / or the catalytic activity of RAD51 protein.

[0103] The present invention also relates to a compound or composition according to the present invention as described herein above for use as a medicament. According to one embodiment, the present invention relates to a compound or composition for use in the treatment of a proliferative disease. According to one embodiment, the present invention relates to a compound or composition for use in the treatment of cancer.

[0104] The present invention also relates to a method of treating a disease in a subject in need thereof, comprising administering to the subject a compound or composition according to the invention as described herein above. According to one embodiment, the disease is a proliferative disease.

[0105] The present invention also relates to the use of a compound or composition according to the invention as described hereinabove, in the manufacture of a medicament, which, according to one embodiment, is for treating a proliferative disease.

[0106] In one embodiment, the proliferative disease is cancer. In a specific embodiment, the cancer is selected from breast cancer, glioblastoma, and multiple myeloma. In one embodiment, the treatment is a first-line, second-line, or third-line cancer treatment.

[0107] According to one embodiment, the compound or the composition is administered to a subject in need thereof simultaneously, sequentially, or separately with at least another active ingredient. In one embodiment, the other active ingredient is administered by simultaneous administration with the compound or the composition. In one embodiment, the other active ingredient is administered by sequential administration with the compound or the composition. In one embodiment, the other active ingredient is administered by separate administration with the compound or the composition.

[0108] According to one embodiment, the compound is not for use in the treatment of a viral disease. According to one embodiment, the compound is not for use in the treatment of HSV, HIV, or CMV. According to one embodiment, the compound is not for use in the treatment of an inflammatory disease. According to one embodiment, the compound is not for use in the treatment of an inflammatory skin disease. According to one embodiment, the compound is not for use in the treatment of an autoimmune disease. According to one embodiment, the compound is not for use in contraception.

[0109] According to one embodiment, the compound is not a follicle stimulating hormone (FSH) antagonist. According to one embodiment, the compound is not a chloride channel or transporter compound (CLC). [Brief description of the drawings]

[0110] [Figure 1] FIG. 1 is a series of histograms showing on the y-axis the degree of oligomerization of RAD51 protein in the presence of one of compounds B-1 through B-3, X-4, and DIDS, respectively, as presented in Example 2 below. [Diagram 2]FIG. 2 is a series of histograms showing the relative binding (%) of single-stranded DNA to RAD51 protein in the presence of one of compounds B-1 through B-3, X-4, DIDS, and BPQ on the y-axis and different amounts of compound (μM) on the x-axis, respectively, as presented in Example 3 below. [Diagram 3] FIG. 3 is a histogram showing on the y-axis the % intracellular HR inhibition in the presence of one of compounds C-3, B-3, and X-2, as presented in Example 5 below. [Figure 4] FIG. 4 is a graph showing cell viability (%) on the y-axis and different amounts of cisplatin (μM) on the x-axis in the presence of cisplatin alone (upper curve with square symbols) or cisplatin with 20 μM of B-2 compound (lower curve with circle symbols), as presented in Example 6 below. [Diagram 5] FIG. 5 is a graph showing cell viability (%) on the y-axis and different amounts of camptothecin (μM) on the x-axis in the presence of camptothecin alone (upper curve with square symbols) or camptothecin with 20 μM of B-2 compound (lower curve with circular dots), as presented in Example 6 below. [Figure 6] FIG. 6 is a graph showing on the y-axis the viability (%) of cells in the presence of one of the compounds B-1 to B-3 or BPQ, as presented in Example 7 below. [Figure 7] FIG. 7 is a graph showing on the y-axis percent viability in the presence of 50 μM of one of the compounds DIDS, B-1 through B-3, or BPQ, as presented in Example 7 below. [Figure 8] FIG. 8 is a histogram showing quantification (%) of cells containing more than 20 Rad51 foci per nucleus following treatment with Compound B-2 20 μM, cisplatin 30 μM, or Compound B-2 20 μM + cisplatin 30 μM, as presented in Example 8 below. EXAMPLES

[0111] The present invention is further illustrated by the following examples.

[0112] Example 1: Synthesis of stilbene compounds material and method material Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-aminobenzenesulfonate) (DADS) was purchased from Alfa Aesar (4,4'-diaminostilbene 2,2'-disulfonic acid, CAS 81-11-8). Other reactants, including 4,4'-diaminostilbene dihydrochloride (CAS 54760-75-7), were purchased from Sigma Aldrich (Bethesda, US). All solvents used were reagent grade. Thin layer chromatography (TLC) was performed on silica-covered aluminum sheets (Kieselgel 60F 254 , MERCK).

[0113] General Method Eluted TLC was developed using UV irradiation (λ = 254 nm) or molybdic acid solution. NMR spectra were recorded on samples dissolved in appropriate deuterated solvents at room temperature using a BRUKER AC300 ( 1 H is 300MHz, 13 C 75MHz) or BRUKER 400 ( 1 H is 400MHz, 13 C, 100 MHz). 1 Tetramethylsilane (TMS) for H, 13Reference materials of deuterated solvent signals relative to C were used. Chemical displacement values ​​(δ) are expressed in parts per million (ppm) and coupling constants in Hertz (Hz). Protons and carbons that were clearly identified are specified. Infrared spectroscopy analyses were recorded on an IRTF Bruker Tensor 27, Vector 22. Low resolution mass spectra were recorded at CEISAM laboratories using a Thermo-Finnigan DSQII quadripolar (NH 3 The high- and low-resolution mass spectra (HRMS in Da) were recorded on an LC-Q-TOF (Synapt-G2 HDMS, Waters) at the IRS-UN center (Mass Spectrometry platform, Nantes).

[0114] Synthesis method General synthesis procedure 1: To a suspension of DADS (1 mmol, 0.37 g) in a mixture of water / dioxane (ratio 10:2), Na 2 CO 3 (5 eq, 5 mmol) was added. The solution became clear and yellow and anhydride (4 mmol) was added along with a catalytic amount of tetrabutylammonium chloride (0.1 eq). After stirring for 2-18 hours, a precipitate formed. This was filtered and diluted with Et 2 O to give the expected pure compound as observed by NMR analysis. For optimization of purity, a second precipitation step (DMSO:Et 2 O) could be carried out.

[0115] General synthetic procedure 2: To a solution of DADS (1 mmol, 0.37 g) in dimethylsulfoxide (DMF) was added N,N-diisopropylethylamine (DIEA) (5 eq). Acid chloride (4 eq) was added and the reaction was heated at 80° C. overnight. After cooling to room temperature, methanol was added until a precipitate formed. This was filtered and eluted with Et2 The precipitate was then dissolved in DMSO and washed with Na 2 CO 3 Add an aqueous solution of Et 2 After precipitation with O, the expected sodium salt was obtained.

[0116] General synthesis procedure 3: To a suspension of DADS (1 mmol, 0.37 g) in a mixture of water / dioxane (ratio 10:2), Na 2 CO 3 (5 eq, 5 mmol) was added. The solution became clear and yellow and chloroformate reagent (2.5 eq) was added. After stirring overnight a precipitate formed. This was filtered and diluted with Et 2 Washing with 20 mL of DMSO provided the expected pure compound as observed by NMR analysis. For optimization of purity, a second precipitation step (DMSO:Et 2 O) could be carried out.

[0117] General synthesis procedure 4: To a suspension of DADS (1 mmol, 0.37 g) in a mixture of water / dioxane (ratio 10:2), add Na 2 CO 3 (5 eq, 5 mmol) was added. The solution became clear and yellow and sulfonyl chloride reagent (2.5 eq) was added. After stirring overnight a precipitate formed. This was filtered and diluted with Et 2 Washing with 20 mL of DMSO provided the expected pure compound as observed by NMR analysis. For optimization of purity, a second precipitation step (DMSO:Et 2 O) could be carried out.

[0118] General Synthetic Procedure 5: CH 2 Cl 2 To a solution of 4,4'-diaminostilbene dihydrochloride (1 mmol, 0.28 g) in 10 mL of ethyl acetate was added N,N-diisopropylethylamine (DIEA) (5 eq). The solution became clear and brown and chloride reagent (2.5 eq) was added. After stirring overnight a precipitate formed. This was filtered and diluted with Et 2Washing with 20 mL of DMSO provided the expected pure compound as observed by NMR analysis. For optimization of purity, a second precipitation step (DMSO:Et 2 O) could be carried out.

[0119] General synthetic procedure 6: To a suspension of DADS (1 mmol, 0.37 g) in water (6 mL), add Na 2 NO 2 (0.14 g, 2.2 eq) in water (2 mL) was added, followed by an aqueous solution of acetic acid (5 eq) in water (2 mL). After stirring for 45 min, the sodium or potassium salt was added (2 eq to 4 eq in 2 mL water) in the presence or absence of a catalytic amount of Cu(I) (0.2 eq). After stirring for 4 h to 3 days, MeOH and Et 2 O was added to give a precipitate, which was filtered and diluted with Et 2 Washing with 20 mL of DMSO provided the expected pure compound as observed by NMR analysis. For optimization of purity, a second precipitation step (DMSO:Et 2 O) could be carried out.

[0120] result The disulfonate stilbene compounds presented in Table 1 above and the comparative non-sulfonated stilbene compounds presented in Table 2 above were synthesized according to General Synthetic Procedures 1-5 described above.

[0121] Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-acetamidobenzenesulfonate) (A-1) was prepared from DADS and acetic anhydride according to general synthetic procedure 1. After purification by precipitation, it was obtained in 62% yield. 1 HNMR (300MHz, DMSO- d6 ):δ ppm 10.03 (s, 2H, 2 x NH), 7.96 (s, 2H, CH=CH), 7.94 (d, 2H, 2 x Har), 7.68 (dd, 2H, 2 x Har), 7.52 (d, 2H, 2 x Har), 2.03 (s, 6H, 2 x CH 3 ). 13C NMR (75MHz, DMSO- d6 ): δ ppm 168.08 (2 x CO), 145.69 (2 x CIVar), 137.29 (2 x CIVar), 129.88 (2 x CIVar), 125.98 (CH=CH), 125.57 (2 x CHar), 118.99 (2 x CHar), 117.74 (2 x CHar), 23.90 (2 x CH 3 ). MS (ESI+) m / z: [M+Na] + = 521.0, MS (ESI - ) m / z: [M-Na] - =475.0. HRMS (ESI + ):[M+Na] C 18 H 16 N 2 O 8 Na 3 S 2 Theoretical value: 521.0041; measured value: 521.0029.

[0122] Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-propionamidobenzenesulfonate) (A-2) was prepared from DADS and propionic anhydride according to general synthetic procedure 1. After purification by precipitation, it was obtained in 76% yield. 1 HNMR (300MHz, DMSO- d6 ): δ ppm 9.63 (s, 2H, 2 x NH), 7.93 (s, 2H, CH=CH), 7.90 (s, 2H, 2 x Har), 7.49 (s, 4H, 4 x Har), 4.12 (q, 4H, 2 x CH 2 ), 1.24 (t, 6H, 2 x CH 3 ). 13 C NMR (75MHz, DMSO- d6): δ ppm 153.43 (2 x CO), 145.71 (2 x CIVar), 137.13 (2 x CIVar), 129.40 (2 x CIVar), 125.88 (CH=CH), 125.68 (2 x CHar), 118.39 (2 x CHar), 117.22 (2 x CHar), 60.04 (2 x CH 2 ), 14.50 (2 x CH 3 ). MS (ESI-) m / z : [M-Na]- = 535.2. HRMS (ESI - ):[M-Na] - C 20 H 20 N 2 O 10 NaS 2 Theoretical value: 535.0457; measured value: 535.0443.

[0123] Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-benzamidobenzenesulfonate) (A-3) was prepared from DADS and benzoic anhydride according to general synthetic procedure 1. After purification by precipitation, it was obtained in 70% yield. 1 HNMR (300MHz, DMSO- d6 ): δ ppm 8.11 (s, 2H, CH=CH), 8.09 (s, 2H, 2 x Har), 7.96 (d, 2H, 2 x Har), 7.69 (d, 2H, 2 x Har), 7.33 (m, 10H, 10 x Har). 13 C NMR (75MHz, DMSO- d6): δ ppm 154.04 (2 x CIVar'), 152.35 (2 x CO), 144.27 (2 x CIVar), 138.64 (2 x CIVar), 132.13 (2 x CIVar), 130.45 (4 x CHar'), 128.67 (2 x CHar), 127.99 (CH=CH), 126.64 (2 x CHar), 122.92 (4 x CHar), 121.57 (2 x CHar), 119.01 (2 x CHar). MS (ESI + ) m / z: [M+Na] + = 676.9. HRMS (ESI + ):[M+Na] C 28 H 20 N 2 O 10 Na 3 S 2 Theoretical value: 677.0252; measured value: 677.0262.

[0124] Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-(4-methoxybenzamido)benzenesulfonate) (A-4) was prepared from DADS and 4-methoxybenzoyl chloride according to general synthetic procedure 2. After purification by precipitation, it was obtained in 44% yield. 1 HNMR (300MHz, CD 3 OD): δ ppm 10.21 (s, 2H, 2 x NH), 8.18 (d, 2H, 2 x Har), 8.05 (s, 2H, CH=CH), 8.02 (d, 4H, 4 x Har), 7.88 (dd, 2H, 2 x Har), 7.58 (d, 2H, 2 x Har), 7.05 (d, 4H, 4 x Har), 3.84 (s, 6H, 2 x CH 3 ). 13 C NMR (75MHz, CD 3OD): δ ppm 164.57 (2 x CO), 161.80 (2 x CIVar'), 145.66 (2 x CIVar), 137.37 (2 x CIVar), 130.27 (2 x CIVar), 129.60 (4 x CHar), 126.77 (2 x CIVar), 126.14 (CH=CH), 125.41 (2 x CHar), 120.35 (2 x CHar), 119.18 (2 x CHar), 113.48 (4 x CHar), 55.35 (2 x OMe). MS (ESI - ) m / z: [M-Na] - = 659.1. HRMS (ESI - ): cald for [M-2Na] 2- C 30 H 25 N 2 O 10 S 2 637.0951; found 637.0932.[M-Na]-C 30 H 24 N 2 O 10 NaS 2 Theoretical value: 659.0770; measured value: 659.0778.

[0125] Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-(4-nitrobenzamido)benzenesulfonate) (A-5) was prepared from DADS and 4-nitrobenzoyl chloride according to general synthetic procedure 2. After purification by precipitation, it was obtained in 70% yield. 1 HNMR (300MHz, DMSO- d6 ): δ ppm 10.70 (s, 2H, 2 x NH), 8.36 (m, 4H, 4 x Har), 8.24 (m, 6H, 6 x Har), 8.08 (s, 2H, CH=CH), 7.91 (dd, 2H, 2 x Har), 7.63 (d, 2H, 2 x Har). 13 C NMR (75MHz, DMSO- d6): δ ppm 163.54 (2 x CO), 149.06 (2 x CIVar), 145.86 (2 x CIVar), 140.41 (2 x CIVar), 136.78 (2 x CIVar), 130.92 (2 x CIVar), 129.23 (4 x CHar), 126.42 (2 MS (ESI + ) m / z: [M+Na] + = 734.9, MS (ESI - ) m / z: [M-Na] - = 688.9. HRMS (ESI + ):[M+Na] + C 28 H 18 N 4 O 12 Na 3 S 2 Theoretical value: 735.0056; measured value: 735.0057.

[0126] Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-((ethoxycarbonyl)amino)benzenesulfonate) (C-1) was prepared from DADS and ethyl chloroformate according to general synthetic procedure 3. After purification by precipitation, it was obtained in 66% yield. 1 HNMR (300MHz, DMSO- d6 ): δ ppm 9.63 (s, 2H, 2 x NH), 7.93 (s, 2H, CH=CH), 7.90 (s, 2H, 2 x Har), 7.49 (s, 4H, 4 x Har), 4.12 (q, 4H, 2 x CH2), 1.24 (t, 6H, 2 x CH 3 ). 13 C NMR (75MHz, DMSO- d6): δ ppm 153.43 (2 x CO), 145.71 (2 x CIVar), 137.13 (2 x CIVar), 129.40 (2 x CIVar), 125.88 (CH=CH), 125.68 (2 x CHar), 118.39 (2 x CHar), 117.22 (2 x CHar), 60.04 (2 x CH 2 ), 14.50 (2 x CH 3 ). MS (ESI - ) m / z: [M-Na] - = 535.2. HRMS (ESI-): [M- Na] - C 20 H 20 N 2 O 10 NaS 2 Theoretical value: 535.0457; measured value: 535.0443.

[0127] Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-((isobutoxycarbonyl)amino)benzenesulfonate) (C-2) was prepared from DADS and isobutyl chloroformate according to general synthetic procedure 3. After purification by precipitation, it was obtained in 67% yield. 1 HNMR (300MHz, DMSO- d6 ): δ ppm 9.65 (s, 2H, 2 x NH), 7.94 (s, 2H, CH=CH), 7.92 (d, 2H, 2 x Har), 7.49 (m, 4H, 4 x Har), 3.87 (d, 4H, 2 x CH 2 ), 1.92 (m, 2H, 2 x CH), 0.94 (d, 12H, 4 x CH 3 ). 13 C NMR (75MHz, DMSO- d6): δ ppm 153.58 (2 x CO), 145.72 (2 x CIVar), 137.169 (2 x CIVar), 129.41 (2 x CIVar), 125.89 (CH=CH), 125.70 (2 x CHar), 118.44 (2 x CHar), 117.25 (2 x CHar), 70.01 (2 x OCH 2 ), 27.55 (2 x CH), 18.90 (4 x CH 3 ). MS (ESI + ) m / z: [M+Na] + = 637.1, MS (ESI - ) m / z: [M-Na] - = 591.0. HRMS (ESI + ):[M+Na] + C 24 H 28 N 2 O 10 Na 3 S 2 Theoretical value: 637.0878; measured value: 637.0870.

[0128] Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-((phenoxycarbonyl)amino)benzenesulfonate) (C-3) was prepared from DADS and phenyl chloroformate according to general synthetic procedure 3. After purification by precipitation, it was obtained in 38% yield. 1 HNMR (300MHz, CD 3 OD): δ ppm 8.11 (s, 2H, CH=CH), 8.09 (s, 2H, 2 x Har), 7.96 (d, 2H, 2 x Har), 7.69 (dd, 2H, 2 x Har), 7.39 (m, 4H, 4 x Har), 7.22 (m, 6H, 6 x Har). 13 C NMR (75MHz, CD 3OD) : δ ppm 154.04 (2 x CIVar), 152.35 (2 x CO), 144.27 (2 x CIVar), 138.64 (2 x CIVar), 132.13 (2 x CIVar), 130.45 (4 x CHar), 128.67 (2 x CHar), 127.99 (CH=CH), 126.64 (2 x CHar), 122.92 (4 x CHar), 121.57 (2 x CHar), 119.01 (2 x CHar). MS (ESI + ) m / z: [M+Na] + = 676.9. HRMS (ESI + ):[M+Na] + C 28 H 20 N 2 O 10 Na 3 S 2 Theoretical value: 677.0252; measured value: 677.0262.

[0129] Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-(phenylsulfonamido)benzenesulfonate) (S-1) was prepared from DADS and benzenesulfonyl chloride according to general synthetic procedure 4. After purification by precipitation, it was obtained in 81% yield. 1 HNMR (300MHz, D 2 O): δ ppm 7.80 (m, 4H, Har), 7.65 (s, 2H, CH=CH), 7.64 (d, 2H, J =8.2 Hz, 2 x Har), 7.60-7.48 (m, 6H, Har), 7.43 (d, 2H, J = 2.44 Hz, 2 x Har), 7.06 (dd, 2H, J = 2.44 Hz and 8.6 Hz, 2X Har). 13 C NMR (75MHz, D 2O): δ ppm 144.06 (2xCIV), 141.11 (2xCIV), 140.19 (2xCIV), 132.18 (2xCHar), 129.11 (4xCHar), 127.91 (2xCHar), 127.61 (2xCIV), 126.47 (4xCHar), 125.78 (CH=CH), 124.40 (2xCHar), 119.66 (2xCHar). HRMS (ESI+):C 26 H 19 N 2 O 10 Na 2 S 4 [MH] - Theoretical value: 692.9718; measured value: 692.9719.

[0130] Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-((4-methylphenyl)sulfonamido)benzenesulfonate) (S-2) was prepared from DADS and para-toluenesulfonyl chloride according to general synthetic procedure 4. After purification by precipitation, it was obtained in 87% yield. 1 HNMR (300MHz, D 2 O): δ ppm 7.68 (m, 4H, 4xHar), 7.64 (s, 2H, CH=CH), 7.64 (s, 2H, CH=CH), 7.64 (d, 2H, 2 x Har), 7.42 (d, 2H, J = 2.4 Hz, 2 x Har), 7.33 (m, 4H, 4 x Har), 7.03 (dd, 2H, J = 8.5 Hz and 2.5 Hz, 2 x Har), 2.35 (s, 6H, 2 x CH 3 ). HRMS (ESI - ):C 28 H 23 N 2 O 10 S 4 Na 2 [M+H] - Theoretical value: 721.0031; measured value: 721.0024.

[0131] Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-(4-nitrophenylsulfonamido)benzenesulfonate) (S-3) was prepared from DADS and 4-nitrobenzenesulfonyl chloride according to general synthetic procedure 3. After purification by precipitation, it was obtained in 81% yield. 1 HNMR (300MHz, D 2 O): δ ppm 8.32 (dl, 4H, 4 x Har), 7.99(dl, 4H, 4xHar), 7.664 (m, 4H, 2 x Har and CH=CH), 7.43 (sl, 2H, 2 x Har), 7.05 (dl, 2H, 2x Har). HRMS (ESI + ):C 26 H 19 N 4 O 14 S 4 Na 2 [M+H] + Theoretical value: 784.9570; measured value: 784.9560.

[0132] (E)-N,N'-(ethene-1,2-diylbis(4,1-phenylene))diacetamide (X-1) was prepared from 4,4'-diaminostilbene dihydrochloride and acetyl chloride according to general synthetic procedure 5. After purification by precipitation, it was obtained in 53% yield. 1 HNMR (300MHz, DMSO- d6 ): δ ppm 10.06 (s, 2H, 2 x NH), 7.54 (m, 8H, 8 x Har), 7.07 (s, 2H, CH=CH), 2.05 (s, 6H, 2 x CH 3 ). 13 C NMR (75MHz, DMSO- d6 ): δ ppm 168.17 (2 x CO), 138.58 (2 x CIVar), 131.95 (2 x CIVar), 126.58 (4 x CHar), 126.47 (CH=CH), 118.98 (4 x CHar), 23.96 (2 x CH 3 ). MS (ESI +) m / z: [M+Na] + = 317.1. HRMS (ESI + ):[M+Na] + The theoretical value of C 18 H 18 N 2 O 2 Na 317.1266; measured value 317.1278.

[0133] Diphenyl(ethene-1,2-diylbis(4,1-phenylene)) (E)-dicarbamate (X-2) was prepared from 4,4′-diaminostilbene dihydrochloride and phenyl chloroformate according to general synthetic procedure 5. After purification by precipitation, it was obtained in 54% yield. 1 HNMR (300MHz, DMSO- d6 ): δ ppm 10.33 (s, 2H, NHCO), 7.57-7.42 (m, 10H, Har), 7.30-7.23 (m, 8H, Har), 7.11 (s, 2H, CH=CH). 13 C NMR (75MHz, DMSO- d6 ): δ ppm 151.54 (2x CIV), 150.41 (2x CIV), 137.81 (2x CIVar), 131.99 (2x CIVar), 129.36 (4xCHar), 126.82 (4xC), 126.52 (CH=CH), 125.39 (2xCHar), 121.89 (4xCHar), 118.48 (4xCHar). HRMS (ESI+):[M+Na] + C 28 H 22 N 2 O 4 Theoretical value of Na is 473.1472; measured value is 473.1471.

[0134] (E)-N,N'-(ethene-1,2-diylbis(4,1-phenylene))dibenzenesulfonamide (X-3) was prepared from 4,4'-diaminostilbene dihydrochloride and phenylsulfonyl chloride according to general synthetic procedure 5. After purification by precipitation, it was obtained in 41% yield. 1HNMR (300MHz, DMSO- d6 ): δ ppm 10.31 (sl, 2H, NH), 7.76 (dd, 4H, J=8.33 Hz and 1.55 Hz, 2 x Har), 7.60-7.54 (m, 6H, 6xHar), 7.38 (d, 4H, J = 8.64 Hz, Har stilb), 7.07 (d, 4H, J = 8.64 Hz, Har stilb), 6.97 (s, 2H, CH=CH). HRMS (ESI + ):[M+Na] + C 26 H 22 O 4 N 2 NaS 2 Theoretical value: 513.0913; measured value: 513.0918.

[0135] (E)-1,2-bis(4-azidophenyl)ethene (X-4) was synthesized using NaN according to general synthesis procedure 6. 3 (2eq) without Cu catalyst. After filtration of the brown precipitate, purification by column chromatography afforded two products, of which the expected (E)-isomer (X-4) was identified. IR (cm-1): υ max = 2111 and 2081 (N 3 ). 1 HNMR (300MHz, DMSO- d6 ): δ ppm 7.64 (d, 4H, CHar), 7.23 (s, 2H, CH=CH), 7.13 (d, 4H, CHar).

[0136] Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-azidobenzenesulfonate) (B-1) was prepared by the general synthesis procedure 6 using NaN 3 (2eq) without copper catalyst. After stirring for 3h 30min, Na 2 CO 3 was added to obtain a pH of about 9. After purification by precipitation, it was obtained in 80% yield. 1HNMR (300MHz, DMSO- d6 ): δ ppm 8.04 (s, 2H, CH=CH), 7.63 (d, 2H, 2 x Har), 7.50 (d, 2H, 2 x Har), 7.12 (dd, 2H, 2 x Har). 13 C NMR (75MHz, DMSO- d6 ): δ ppm 147.07 (2 x CIV), 136.99 (2 x CIV), 131.82 (2 x CIV), 127.05 (2 x CHar), 126.50 (CH=CH), 119.55 (2 x CHar), 117.32 (2 x CHar). MS (ESI - ) m / z: [M-Na] - = 442.9. HRMS (ESI - ):[M-Na] - The theoretical value of C 14 H 8 N 6 O 6 NaS 2 ; 442.9844 Actual value 442.9858.

[0137] Sodium (E)-6,6'-(ethene-1,2-diyl)bis(3-cyanobenzenesulfonate) (B-2) was prepared from DADS using NaCN (0.2 g) and CuCl (0.05 g) as catalysts according to general synthetic procedure 6. After purification by precipitation, it was obtained as a brown powder in 59% yield. 1 HNMR (300MHz, DMSO- d6 ): δ ppm 8.27 (s, 2H, CH=CH), 8.07 (d, 2H, 2 x Har), 7.88 (dd, 2H, 2 x Har), 7.77 (d, 2H, 2 x Har). 13 C NMR (75MHz, DMSO- d6): δ ppm 146.59 (2 x CIV), 138.83 (2 x CIV), 132.64 (2 x CHar), 130.51 (2 x CHar), 129.50 (2 x CH=CH), 126.64 (2 x CHar), 118.62 (2 x CN), 109.25 (2 x CIV). MS (ESI - ) m / z: [M-Na] - = 411.

[0138] Potassium (E)-6,6'-(ethene-1,2-diyl)bis(3-iodobenzenesulfonate) (B-3) was prepared from DADS using KI (5 eq, 0.83 g) and CuI (0.1 eq, 0.02 g) as catalysts according to general synthetic procedure 6. After purification by precipitation, it was obtained as a brown powder in 41% yield. 1 HNMR (300MHz, D2O): δ ppm 8.23 ​​(d, J = 1.4 Hz, 4H, CHar), 7.92 (dd, J = 1.4 Hz and J = 8.2 Hz, 4H, CHar), 7.84 (s, 2H, CH=CH), 7.61 (d, J = 8.3 Hz, CHar (5)). 13 C NMR (75MHz, D 2 O): δ ppm 140.1 (2 x CIV), 140.65 (2 x CHar), 135.52 (2 x CHar), 134.07 (2 x CIV), 128.9 (2 x CHar),127.9 (CH=CH), 92.37 (2 x CI). MS (ESI - ) m / z: [M-2K+Na] - = 612.8; [M-2K+H] - = 590.8; [MK] - = 628.8. HRMS (ESI): [MK] - C 14 H 8 O 6 N 2 Theoretical value of K is 628.7489; measured value is 628.7496.

[0139] Example 2: In vitro assay - RAD51 oligomerization by SDS-PAGE gel (cross-linking) material and method The reaction was carried out at room temperature. 2.5 μM HsRad51 was incubated with the indicated concentrations of test compound in 20 mM sodium phosphate, 50 mM NaCl, 1 mM MgCl 2 The cross-linking reaction was initiated by the addition of disuccinimidyl suberate (50 μm) and allowed to proceed for 30 min. The reaction was quenched by the addition of Tris-HCl (pH 7.5, final concentration 50 mm) followed by incubation for 15 min. Products were then separated by 12% SDS polyacrylamide gel electrophoresis and detected by Western blot using a mouse monoclonal anti-Rad51 antibody (NeoMarkers) and an anti-mouse Alexa-Fluor 700-conjugated secondary antibody (Molecular Probes, Eugene, OR). Crosslinked dimers were quantified using an Odyssey Infrared Imaging System scanner (Li-Cor Biosciences) (Martinez, SF et al., "Targeting human Rad51 by specific DNA aptamers induces inhibition of homologous recombination", Biochimie, December 2010, Vol. 92, No. 12, pp. 1832-1838).

[0140] result SDS-PAGE electrophoresis after cross-linking maintains and fixes the RAD51 protein complex, thus allowing visualization of the degree of oligomerization (polymerization) of the RAD51 protein. Thus, the effects of RAD protein inhibitors, B-1, B-2, B-3, DIDS, and X-4, on the key steps of the recombinase activity of this RAD51 protein have been evaluated. The results are presented in Figure 1.

[0141] These results show that in the presence of disulfonatostilbenes B-1, B-2, and B-3, a significant reduction in the oligomerization state of RAD51 was observed. The oligomeric form is very strongly reduced for B-2, which contains two cyano groups. This reduction induced by disulfonatostilbenes B-1, B-2, and B-3 is within the same order of magnitude compared to that observed with the reference compound DIDS. It is the applicant's understanding that DIDS and B-1 to B-3 prevent oligomerization by acting at the interface between monomers of the RAD51 protein.

[0142] Notably, in contrast to B-1, the comparative compound X-4, which does not contain a sulfonate group, has no effect on the oligomerization of RAD51 protein, suggesting that the two sulfonate groups are essential for the inhibitory properties of RAD51 protein.

[0143] Thus, the disulfonatostilbenes of the present invention are potent RAD51 protein inhibitors, as evidenced by the prevention of RAD51 protein oligomerization.

[0144] Example 3: In vitro assay - RAD51 nucleofilament formation by interferometry material and method Binding of Rad51 to ssDNA was monitored and quantified by Bio-Layer Interferometry (BLI) technology using the BLItz platform (ForteBio Inc.). The procedure included four steps: (1) 1× PBS, 10 mM MgCl 2and a baseline in buffer containing 0.0075% Tween 20 was recorded for 20 seconds; (2) Rad51 loading and binding was measured for 40 seconds using 1.5 μM Rad51 and 1 mM ATP in buffer A; (3) Rad51 dissociation from ssDNA was recorded for 20 seconds in buffer A; (4) ssDNA biosensor was regenerated using a 50 mM NaOH bath. Steps 1, 3, and 4 used 250 μL of solution in a 0.5 mL tube, except for step 2, which used a 4 μL drop of protein sample. Measurement parameters were as follows: room temperature measurement and stirring speed 2200 rpm. To investigate the effect of CSB and analogs on Rad51 ssDNA binding, the molecules were included in the protein sample from step 2 at the indicated concentrations and incubated for 5 min before loading onto the biosensor (Normand, A. et al., "Identification and characterization of human Rad51 inhibitors by screening of an existing drug library.", Biochemical Pharmacology, October 2014, Vol. 91, No. 3, pp. 293-300.).

[0145] result A test based on interferometry (Blitz®, Fortebio) allows the measurement of the kinetics of binding of the RAD51 protein with single-stranded DNA immobilized on a biosensor support in the presence or absence of inhibitors. The effect of compounds B-1, B-2, B-3, BPQ, and X-4 on the formation of RAD51 nucleofilaments has been evaluated in the presence of increasing doses of inhibitors. The results are presented in FIG. 2.

[0146] These results show that disulfonatostilbenes B-1, B-2, and B-3 inhibit the formation of RAD51 protein nucleofilaments. The formation of nucleofilaments is very strongly reduced in B-2, which contains two cyano groups. The activity of B-1, B-2, and B-3 is comparable to or even better than one of the reference substances BPQ. The activity of B-1, B-2, and B-3 is comparable to one of the reference substances DIDS at low doses (5 μM). B-2 has the same effect compared to DIDS at high doses (50 μM).

[0147] These results also show that the comparative compound X-3 has no inhibitory effect on the RAD51 protein at any dose.

[0148] Thus, the disulfonatostilbenes of the present invention are potent RAD51 protein inhibitors, as evidenced by the prevention of RAD51 protein nucleofilament formation.

[0149] Example 4: In vitro assay - DNA invasion by D-loop assay material and method Labeled 100-ssDNA ** (1 μM) in 20 mM Tris-HCl (pH 8), 1 mM ATP, 1 mM DTT, 1 mM CaCl 2 The reactions were incubated with 0.5 μM Rad51 in the presence or absence of the indicated amounts of the corresponding molecules in 10 μL of standard reaction buffer containing 10 mM Tris-HCl (pH 8), 10 mM MgCl 2 for 20 min at 37 °C. The reactions were initiated by the addition of supercoiled pPB4.3 DNA (200 μM in bp). After incubation for 30 min at 37 °C, the reactions were stopped with stop solution (10 mM Tris-HCl (pH 8), 10 mM MgCl 2 2The reaction mixture was stopped and deproteinized with 100-ssDNA (1% SDS, 1 mg / mL proteinase K). The reaction mixture was further incubated at 37°C for 15 min. After addition of 5x loading dye (0.05% bromophenol blue, 8% glycerol, 1 mM EDTA), the reaction products were separated by electrophoresis on a 1% agarose gel. Electrophoresis was performed at 100 V for 2 h in 0.5XTAE buffer (20 mM Tris, 10 mM acetic acid, and 1 mM EDTA). The labeled products (100-ssDNA ** and D-loop) were visualized and quantified by detection of IRD-700 dye using the 700 nm infrared fluorescence detection channel of an Odyssey Infrared Imager (LI-COR) (Normand, A. et al., Biochemical Pharmacology, October 2014, Vol. 91, No. 3, pp. 293-300.).

[0150] result The "D-loop" in vitro test allows the quantification of the formation of the strand exchange structure (D-loop) and thus the determination of the inhibitory power of a molecule during the strand exchange step of the recombinase mechanism. Next, the IC2000 was used to measure the inhibition of the RAD51 protein in the presence of compounds A-1 to A-5, B-1 to B-3, B-1 to B-3, and DIDS. 50 The results are presented in Table 3 below. [Table 3] Table 3

[0151] These results show that disulfonatostilbenes A-3, A-4, C-3, S-1, S-2, B-1, B-2, and B-3 have relatively low IC 50 Thus, they are potent inhibitors of strand-exchange structure (D-loop) formation. Disulfonatostilbenes A-4, C-3, and B-2 have very low IC values ​​within the same order of magnitude of the DIDS reference compound. 50It has a high molecular weight and is therefore particularly efficient in preventing strand exchange steps.

[0152] Thus, the disulfonatostilbenes of the present invention are potent inhibitors of the RAD51 protein, as evidenced by DNA invasion.

[0153] Example 5: Cellular assay - Effects on intracellular homologous recombination (HR) material and method After transfection with pCMV-HA-I-SceI plasmid and incubation for 72 hours, cells were harvested in PBS and 50 mM EDTA, pelleted, and fixed with 2% paraformaledhyde for 20 minutes. The percentage of cells expressing GFP was scored by FACS analysis using a Cytoflex (Beckman Coulter). At least three independent experiments were performed, and the expression of HA-I-SceIb was verified each time by Western blot with rabbit anti-HA antibody (Abcam).

[0154] result The recombinant fibroblast cell line RG37 is used to confirm the in vitro results (examples 2-5) in a cellular context. This cell line contains two non-functional GFP cassettes, one of which has a unique cleavage site inducible by transfection of a restriction enzyme. After a cleavage equivalent to a double-strand break (DSB) in DNA, homologous recombination (HR) leads to the reconstitution of the complete GFP cassette, thus making the cells fluorescent. The intracellular HR rate is then determined by flow cytometric measurement, measuring the levels of HR in cells after treatment with compounds C-3, B-2, and X-2. The results are presented in FIG. 3.

[0155] Thus, these results confirmed the in vitro results, since significant inhibitory effects on HR were obtained with disulfonatostilbenes C-3 and B-2, whereas the comparative compound X-2, which does not contain a sulfonate group, did not produce any inhibition (Figure 3).

[0156] Thus, the disulfonatostilbenes of the present invention are potent RAD51 protein inhibitors, as demonstrated by intracellular homologous recombination.

[0157] Example 6: Cellular assay - Sensitization with anti-cancer drugs material and method Clonogenicity tests are performed on several model cell lines derived from cancer, including prostate cancer (DU-145). Cells are exposed to DNA-targeting anticancer drugs (cisplatin or camptothecin). The number of damaged cells is counted and seeded in triplicate culture plates in the presence or absence of B-2. After approximately 2 weeks of incubation at 37°C, the number of clones formed is counted. Each cell that is able to repair its damage then forms a clone. After counting, the percentage of survival is calculated relative to the control condition, where 100% are considered to be alive.

[0158] result The main interest of RAD51 protein inhibitors is the sensitization of chemo- or radio-resistant cancer cell lines. Clonogenic potential was evaluated in DU-145 cell lines treated with cisplatin treatment in the presence or absence of disulfonatostilbene B-2 (RAD51 protein inhibitor). The results are presented in Figure 4 and Table 4 below. [Table 4]

[0159] These results show that cell viability is higher in the presence of cisplatin alone (upper curve with square symbols) than in the presence of cisplatin with 20 μM of B-2 compound (lower curve with circle symbols) regardless of the amount of cisplatin (Figure 4). Thus, B-2 indeed acts as a sensitizing agent for cancer cell lines that complements the antineoplastic activity of cisplatin. This action also increases the IC of growth inhibition of cisplatin. 50 This is also evidenced by the approximately 40% decrease in the presence of B-2 (Table 4).

[0160] The same experiment was performed using camptothecin treatment, which is known to induce DNA damage that is repaired by RAD51-mediated homologous recombination.

[0161] The DU-145 cell line was treated with camptothecin in the presence or absence of disulfonatostilbene B-2 (RAD51 protein inhibitor) and the survival of clonogenic cells was determined, the results of which are shown in Figure 5 and Table 5 below. [Table 5]

[0162] These results show that cell viability in the presence of camptothecin alone (upper curve with square dots) is higher than in the presence of 20 μM of B-2 compound and camptothecin (lower curve with circular symbols), regardless of the amount of camptothecin (Figure 5). Thus, B-2 indeed acts as a sensitizer for cancerous cell lines that complements the antineoplastic activity of camptothecin. This action also increases the IC of camptothecin growth inhibition. 50 This is also evidenced by the approximately 90% reduction in the presence of B-2 (Table 5).

[0163] Thus, the disulfonatostilbenes of the present invention are potent sensitizing agents and are therefore useful in the treatment of proliferative diseases.

[0164] Example 7: Cellular Assay - Cytotoxicity material and method Prostate cancer cells DU145 (cancer) and fibroblast cells were cultured in minimum essential medium (RPMI1640 medium) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin at 37 °C for 24 h at 5% CO. 2 , and grown at 37°C.

[0165] The treated molecules were dissolved and stored in DMSO at a concentration of 20 mM.

[0166] DU145 cells and fibroblasts were cultured at 5 x 10 in 100 μl of medium in a standard 96-well plate. 3 The plates were incubated at 37 °C for 24 h at 5% CO 2 The mixture was incubated at 37° C. for 24 hours in an atmosphere of 0.1%. The molecule solutions diluted in DMSO were serially diluted from 60 μM to 0.02 μM with RPMI 1640 medium and added in quadruplicate to 100 μl of RPMI.

[0167] After 72 hours of incubation, 20 μl of 5 mg / ml-1 MTT was added to each well according to the manufacturer's instructions. After 2 hours of incubation at 37°C, the medium was removed and 200 μl of DMSO was added to each well to dissolve the formazan crystals. The absorbance of each well at 540 nm was then monitored using an EnSpire multimode plate reader. The absorbance of each well was expressed as a percentage of the control (wells with untreated cells) and was calculated as the 50% inhibitory concentration (IC 50 ) was decided. IC 50 Values ​​are defined as the concentration of drug that results in 50% growth inhibition. Results represent the mean of at least three experiments. All data are expressed as the mean ± standard deviation. IC 50 Values ​​were calculated by linear regression analysis of concentration-log response curves using the software GraphPad InStat 3.0.

[0168] result The DU245 prostate cancer cell line was used to evaluate and compare the toxicity of small molecule stilbene compounds of the present invention B-1 to B-3 and the reference compound BPQ.

[0169] Compounds B1-B3 had no cytotoxic effect at 50 μM for 3 days on DU-145 cells, whereas BPQ produces high cytotoxicity (low cell viability) in cells at the same incubation time and concentration (Figure 6). Indeed, BPQ produces 90% cell mortality, whereas the stilbene derivatives of the invention do not affect cell viability (>95% cell viability). The same results are obtained with fibroblast cells (Figure 7).

[0170] Studies carried out on fibroblasts (FIG. 7) also show that DIDS, contrary to the stilbene derivatives of the invention, has a cytotoxic effect at 50 μM for 3 days.

[0171] Example 8: Cellular assay - Quantification of Rad51 foci material and method DU-145 cells were cultured at 5 × 10 per well in medium (RPMI) supplemented with 10% fetal bovine serum (Gibco Invitrogen). 3 Cells were grown on glass coverslips (Life Technologies) at a density of 100 / well. Cells were treated with 30 μM cisplatin in the presence or absence of 20 μM RAD51 inhibitor B-2 for 1 h, and then the medium was replaced with molecule-free medium. After 18 h, cells were washed with PBS for 5 min and fixed with absolute ethanol for 25 min.

[0172] Fixed cells were incubated in PBS buffer containing anti-RAD51 antibody diluted 1:1000 for 1 h at room temperature. After washing three times with PBS for 5 min, slides were incubated in PBS solution containing secondary antibody conjugated with AlexaFluor 555 (dilution 1:0000) for 45 min in a dark room. Slides were washed three times with PBS, counterstained and mounted with ProLong Antifade with DAPI (4',6-diamidino-2-phenylindole) (Life Technologies), and coverslipped. The slides were observed under a confocal microscope (Nikon A-1RSi, Minato-ku, Tokyo, Japan) and an epifluorescence microscope (Nikon Eclipse E800). Images were recorded using NIS Element software (Version 3.6, Nikon, Tokyo, Japan). Quantification of RAD51 foci was processed with a program from Fiji-ImageJ software (NIH, Bethesda, Maryland, USA).

[0173] result The DU145 prostate cancer cell line was used to assess the levels of RAD51 foci following cisplatin treatment in the presence or absence of B-2.

[0174] Treatment with cisplatin alone induces high levels of RAD51 foci, but this level is reduced when treatment is combined with 20 μM B-2. Following quantification of the number of RAD51 foci per cell, the number of cells containing more than 20 RAD51 foci was determined.

[0175] The histogram graph in Figure 8 confirms that the level of RAD51 foci mediated by 30 μM cisplatin is reduced (by about 20%) by B-2, indicating that B-2 can reduce RAD51 foci formation after cisplatin-mediated DNA damage.

Claims

1. 1. A pharmaceutical composition for use in the treatment of a proliferative disease, comprising a compound of formula (I): 【Chemistry 1】 or a pharma- ceutically acceptable salt thereof, wherein R ０Ａ and R ０Ｂ is independently selected from hydrogen, lithium, sodium, and potassium; R Ａ and R Ｂ are the same and are selected from azide, cyano, and halide; Pharmaceutical compositions.

2. 2. The pharmaceutical composition of claim 1, wherein the halide is iodine.

3. R ０Ａ and R ０Ｂ The pharmaceutical composition according to claim 1 or 2, wherein both are sodium.

4. R ０Ａ and R ０Ｂ The pharmaceutical composition according to claim 1 or 2, wherein both are potassium.

5. The compound of formula (I) 【Table 1】 The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition is selected from the group consisting of

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the proliferative disease is cancer.

7. The pharmaceutical composition of claim 6, wherein the cancer is breast cancer, glioblastoma, or multiple myeloma.

8. The pharmaceutical composition according to any one of claims 1 to 7, further comprising at least another active ingredient.

9. The pharmaceutical composition according to claim 8, wherein the at least another active ingredient is an antineoplastic active ingredient.

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